v3 Nov 2024

Author instructions
 Please fill in the template below to generate your manuscript.
 We strongly recommend saving a new copy of this template and using one version
as a guide.
 Prior to drafting your manuscript, please read our Information for authors: STAR
Protocols.
 The Protocol should be written:
o Spelling and use of periods and commas in numbers should conform to
American English usage.
o In the present tense and using active and imperative verbiage throughout.
o For example
 Default Active Voice: We fill the bottom of a cryo-mold with OCT Cryo-
embedding medium.
 Imperative Active Voice: Fill the bottom of a cryo-mold with OCT Cryo-
embedding medium.
 A Protocol should be limited to one or more sections of related experimental steps.
Manuscripts containing multiple unrelated protocols will be asked to submit them
separately or remove unrelated protocols.
 Replace any text within [square brackets] with text relevant for your protocol and
delete the brackets.
 Instructions for preparing your manuscript are provided as Editor Instructions in red
italics text. Delete before submitting your manuscript.
 Text boxes outlined in red inserted in this document serve as Example Text. Delete all
text boxes before submitting your manuscript.
 Please keep all sections (identified Optional sections can be deleted if not applicable).
 NOTE: Manuscripts lacking the Key Resources Table, Troubleshooting, Limitations or
Expected Outcomes sections will not be sent out for review and returned to the
author for revision.

Manuscripts without an associated primary research manuscript

 For manuscripts without an associated primary research manuscript, you are required
to provide sufficient raw and analysed data sets derived from the use of the protocol
to allow for reviewers to evaluate the reproducibility and validity of the protocol.
 This can include, but not limited to replicated control sets, imaging standards,
deposited data sets, antibody validation, code repositories.
 Please consult our policy here for more information.
 Please contact our editorial team if you have any questions.

Protocols containing code.

 If your protocol contains code, please follow these guidelines:
 o For small snippets of code, please use a monospace font and indented text as
shown below:
>save.image("STAR Protocols.Rdata")

o For larger code segments, please put the code in an in-line text box in Word
exactly where the code should appear (see example below). Please consult this
walk through on how to add in-line text box.
o Code should be inputted as black text. If you wish to use code with colored text, it
must be submitted as a figure, but users will not be able to copy/paste the code.

>if(!require(devtools)){
> install.packages("devtools") # If not already
installed
>}
>devtools::install_github('Sbuttery/protocols')

o A single section of code may not be longer than three pages. If it exceeds this
limit, submit the code as a supplementary file.
o You can find an example of a computational protocol here.

Tables
 Include a title for each table.
 Include table callouts in the text close to where it is first cited (e.g., Table 1)
o Recipe tables DO NOT require a call out or legend.
 Prepare all tables using the table function in a word-processing program.
 Do not include colored text, indented text, shading, or panels in the table.
 Please ensure that all tables and cell lines and borders are visible.

Figures
 All figures should be uploaded separately as high-resolution (300 dpi) files during
submission. We accept TIFF, PDF, CDX (ChemDraw), or JPEG files for figures.
o Please refer to the Cell Press Digital Image guidelines for more information.
o NOTE: All microscopy images must include appropriate scale bars.
 If your protocol re-uses any part of a figure from previous manuscripts from your lab,
you should be sure to obtain permission to use or adapt the figure from the copyright
holder. Elsevier (including Cell Press) authors retain the right to use or re-use portions
or excerpts in other works; see
https://www.elsevier.com/about/our-business/policies/copyright/personal-use.
 Avoid the use of direct re-use of unmodified figures from your research manuscripts.
 The author should also include an image attribution line in the figure legend, e.g.,
"Figure reprinted and adapted with permission from Smith et al., 2015".
 For figures such as screen shots of computer programs or chemical structures:
 o In the file name, include “Fixed_Image_XXX”. Replace the XXX with name of
file.
o Indicate in the text where the screen shot is to be placed using the following
notation: [*Insert Fixed_Image_XXX here]

Supplemental materials
 Note that the file size limit for supplemental material is 150MB. If needed, consider
dividing the data into separate submission items, each with a descriptive title and
"Related to" information in the main manuscript file.
 The total size of all Supplementary Files is 1.5GB
 Supplementary material should be referenced in the text at the appropriate location
in the format of:
o Figure/Table/Video S1: One-sentence descriptive title for the item, related to
Step
 Please refer to the Supplemental information guidelines: Cell Press

Video Files
 Smart phone camera videos are usually adequate.
 Use landscape not portrait view.
 Video files should be less than 150MB and be in .MP4 format.

Resource Availability and Data Repositing

 For depositing of data associated with your protocol must follow the Cell Press
guidelines under Data and Code Availability.

 If you have produced a dataset with your method: consider submitting a data
article to Data in Brief. This is an Open Access data journal which allows you to
receive credit and citations for the description of your dataset.
 Please ensure that you have archived a “version of record” of your GitHub repository
or code at Zenodo: https://guides.github.com/activities/citable-code/. This will
provide you with a DOI at Zenodo.

Submitting your manuscript

 Please submit your manuscript to https://www.editorialmanager.com/star-
protocols/default.aspx. Be sure to upload the Word version of this manuscript at
submission.
 Before submitting your protocol, please delete the Author instructional pages, all
italicized instructions and Editor’s Note text boxes from your final manuscript version
before submitting. You can see a reference of a properly formatted manuscript here.
[Title]
EDITOR INSTRUCTIONS
 Choose a concise title that includes the name of the method/technique, the purpose of the
protocol, and the model system or organism.
 The title should be no more than 145 characters total (including spaces).

Example Title:
Protocol to study projection-specific circuits in the basal ganglia of adult mice using viral vector
tracing, optogenetics, and patch-clamp technique.

[Author (First Name Surname),1,2,3,* Author (First Name Surname),1,2 …and Author (First Name Surname)1,4,**]
1
Affiliation, including city, state, postal code, and country.
2
Optional footnotes such as current addresses, equal contributions and/or senior authorship
3
Technical contact
4
Lead contact
*Correspondence: Author’s email
**Correspondence: Author’s email]

EDITOR INSTRUCTIONS
 Explanations of the roles and responsibilities of the corresponding author(s) and lead
and technical contacts can be found in the authorship section of the information for authors.
 Please include email addresses for both the lead and technical contact in the Resource
availability section. Please note that:
o The technical contact must match the name/contact in the Resource availability
statement.
o You may designate only ONE lead contact.
 Please check the author roles carefully; changes after acceptance will delay publication and
will require approval of the editor and written agreement among all affected authors.
Changes after publication will require approval of the editor, written agreement among all
affected author, and a published Correction.

Summary
EDITOR INSTRUCTIONS
 The summary is a paragraph no longer than 80 words and written in the active voice and
present tense.
 Any background information should be limited to one sentence. The summary should focus on
the details of the major steps of the protocol, techniques involved, and model organisms used.
 Avoid the use of the word “method” in your summary. Acceptable substitutions are
“protocol”, “technique”, “assay” or “approach”. Do NOT include references in the summary.
 Summaries should include the statement as the last sentence: For complete details on the use
and execution of this protocol, please refer to X et al. (1).
 o Only reference published, peer-reviewed and associated manuscripts in this
statement. If you do not have an associated published manuscript, do not include this
statement, and make note in your submission cover letter.
o This statement does not count towards the 80-word limit for the Summary.

Example Summary (delete before submitting your manuscript):

Pioneer transcription factors (TF) can directly establish higher-order chromatin interactions to instruct gene
transcription. Here, we present a protocol for capturing TF-mediated 3D chromatin interactions using
affinity tag-based BL-HiChIP. We describe steps for constructing flag-tagged TF, performing BL-HiChIP, and
library preparation. We then detail procedures for sequencing and data analysis. This protocol has potential
application in 3D chromatin analysis centered on any specific TF in the absence of optimal antibodies.

For complete details on the use and execution of this protocol, please refer to X et al.1

Graphical abstract
EDITOR INSTRUCTIONS
 The GA is a graphical description of the protocol that highlights the time for each major step.
Click here to see examples. We strongly encourage you to submit a GA with your initial
submission, but it is not required; however, it will be required if your paper is accepted.
 We have created a graphical abstract template (**if the link is not working, please copy and
paste this link to your web browser: https://www.cell.com/pb-assets/journals/EM/STAR
%20Protocols/STAR_GA_Template_SP_Finalized-1694695050470.pptx) to help you create a
GA. (Click here for the Adobe Illustrator version and follow these instructions.)
 Programs such as Biorender may also be used with licensing and citation to create the
Graphical Abstract.

Before you begin

EDITOR INSTRUCTIONS
 Describe the protocol for a specific usage case (ideally what was done in your associated
research paper). You may also describe alternate applications of the protocol.
o For example: “The protocol below describes the specific steps for using HeLa cells.
However, we have also used this protocol in HEK293 cells and NIH3T3 cells.”
 Keep background information related to the protocol to a minimum. Introductory text should
not exceed 1 page.
 Provide instructional steps for what needs to be set up or prepared before a researcher begins
the protocol. Please use numbering (up to three levels: 1,2,3; a,b,c; i,ii,iii) to list steps. Do not
use bullet points in this section.
o Avoid including more than 15 sub-steps for an individual step.
 All materials recipes should be listed in the Materials and Equipment Setup section.
Do not place them here.
  For explanatory text related to the steps, please use the following callouts that can be inserted
after the relevant numbered step.
o Call outs are not to contain instructional steps.
o Optional: [Call out optional steps in this format; optional steps are not numbered.]
o Note: [Please include general notes or explanations as a note. Notes are not
numbered and should be near the relevant step.]
o Pause point: [If relevant, list potential pause points with time intervals in this format;
pause points are not numbered.]
o CRITICAL: [when applicable, note steps that are critical for success of the protocol;
critical steps are usually not numbered.]

Institutional permissions (if applicable)

EDITOR INSTRUCTIONS
 Any experiments on live vertebrates or higher invertebrates must be performed in accordance
with relevant institutional and national guidelines and regulations.
 In the before you begin section, a statement identifying the committee approving the
experiments and confirming that all experiments conform to the relevant regulatory standards
must be included.
o In addition, you should provide a statement reminding readers that they will need to
acquire permissions from the relevant institutions.

[Preparation one]
EDITOR INSTRUCTIONS
 Replace the Preparation One description with your own text. For example: Primer design, Cell
Culture, Sample preparation.
 Steps should be written in the active voice, present tense and in an instructional style.
 Include figures and/or videos to help guide critical steps. Include figure callouts in the text
(e.g., Figure 1) to indicate where figures should appear.
 Timing must be included with each major step. It is also possible to include additional timing
notes for sub steps, but only if the total time for the major step is included. For time ranges
that are difficult to define, you may use ranges or “variable”.

Timing: [s, min, h, days, or weeks.]

1. [First step of preparation one. Use numbers to order steps.]

Step instruction

2. [Second step of preparation one]

a. [Sub-steps under first step. Use letters for ordering of sub-steps.]

CRITICAL: [when applicable, note steps that are critical for success of the protocol]

b. [Sub-step two, as needed.]

 i. [Third-level sub-steps under second-level sub-steps. Use lowercase Roman
numerals for ordering.]
ii. [Third-level sub-step two, as needed.]

[Preparation two]

Timing: [s, min, h, days, or weeks]

EDITOR INSTRUCTION: Step numbers between Preparations are continuous.

3. [First step of preparation two. Use numbers to order steps.]

a. [Use letters for ordering of sub-steps.]
b. [Sub-step two, as needed.]
4. [Second step of preparation two]

Step Example:

1. Remove medium from culture dish by gentle aspiration.

a. Wash cells twice with sterile 1X PBS
b. Add enough trypsin-EDTA to completely cover the cells.
c. Place them at 37 °C for 2 min.
d. Check under the microscope if the cells have detached from the
plate.
Critical: It is important to ensure that all cells have detached before
proceeding to the next step.

2. Add 4 mL of complete medium to stop the trypsin-EDTA reaction.

a. Collect all the liquid in a sterile tube.
b. Centrifuge the cell suspension for 5 min at 400-600 g at RT.
c. Remove the trypsin-EDTA solution by aspiration.
d. Mix the cells with fresh medium containing serum.
Note: You can resuspend cells in 1 mL of fresh medium.

3. Determine cell concentration using a hemocytometer or Burker camera.

Note: In this step you can check cell viability, cell number, and
concentration

Key resources table

EDITOR INSTRUCTIONS
 The Key Resources Table (KRT) highlights key reagents, resources, and equipment used in
the protocol. Include vendor/manufacturer and the model/catalog number for each
reagent or resource.
o Whenever possible, use RRIDs as the identifiers for the items used in the protocol
(see https://scicrunch.org/resources).
  To create the key resources table using Word, please use this table template
(http://www.cell.com/pb-assets/journals/research/cell/methods/table-template1.docx).
 DO NOT CHANGE THE HEADINGS IN THE KRT
 For reagents/resources that are critical for the success of this protocol, please provide
links to the corresponding vendor sites.
 Please note that the heading "Other" is for items not covered by other headings and all
equipment (microscopes, flow cytometers, etc).
 Please do not leave blank spaces on the key resources table (use n/a if there is no
information to include). Also, note that only the standardized subheadings can be used
within the table.
 The KRT can be inserted into the manuscript (preferred method), or alternatively uploaded
as a separate document.
Materials and equipment setup (optional)
EDITOR INSTRUCTIONS
 This section is for providing additional information regarding equipment setup, details about
custom software used, and notes on alternative materials and equipment. This optional
section is a complement to the key resources table.
 ALL EQUIPMENT SHOULD BE LISTED IN THE KRT UNDER “OTHER”, NOT HERE.
 All recipes should be in this section; these tables do not require legends or call outs and can
stay in this section.
 Please use bullet points (up to three level) to list items. Do not use numbering (1,2,3; a,b,c;
i,ii,iii) in this section.
 Recipes with ≥3 ingredients should be presented as tables. All solutions should include storage
conditions (temperature and maximum store time).

Materials Examples

[Name of solution/buffer/medium]
Reagent Final concentration Amount
Component A (100 µM) X 1 mL
Component B (100 µM) Y 1 mL
10 × ligase buffer Z 5 mL
T4 polynucleotide kinase Z 1 mL
ddH2O n/a 42 mL
Total n/a 50 mL
[Note on storage conditions e.g. Store at -4C for up to 3 months]

CRITICAL: [Mark any chemicals or reagents that are harmful or toxic with a brief explanation of the
hazard, and state clearly how to take precautions when handling these agents.]

Alternatives: [Mention any known alternative materials or equipment in this format.]

Step-by-step method details

EDITOR INSTRUCTIONS
  This section lists the major steps and provides step-by-step details and timing for each major
step.
 Please use continuous numbers for this section through Major Steps. Do not continue
numbering from the Before You Begin section.
 Please use numbering (up to three levels: 1,2,3; a,b,c; i,ii,iii) to list steps. Do not use bullet
points in this section.
o Avoid including more than 15 sub-steps for an individual step.
 DO NOT number the Major Steps.
 When describing the protocol steps, write in the present tense and use active and imperative
verbiage throughout. See authors instructions page for an example.
 For steps using manufacturer's protocols, please include link(s) to the protocols.
 Whenever possible, include links between troubleshooting sections and major steps.

[Your major step one]

EDITOR INSTRUCTIONS
 Your major step title should be descriptive. Avoid short titles such as “Cell culture”, “Western
blot”, or “qPCR”.
 Include a brief description (no more than 2 sentences) about what this major step
accomplishes; this will help other researchers repeat and troubleshoot the protocol.
 Timing must be included with each major step. It is also possible to include additional timing
notes for sub steps, but only if the total time for the major step is included. For time ranges
that are difficult to define, you may use ranges or “variable”.

Timing: [s, min, h, days, or weeks]

1. [First step of major step one. Use numbers to order steps.]

a. [Sub-steps under first step. Use letters for ordering of sub-steps.]
b. [Sub-step two, as needed.]
i. [Third-level sub-steps under second-level sub-steps. Use Roman numerals for
ordering.]
ii. [Third-level sub-step two, as needed.]
2. [Second step of major step one]

[Your major step two]

Timing: [s, min, h, days, or weeks]

[Include a brief description about what this major step accomplishes. This will help other researchers
repeat and troubleshoot the protocol.]

3. [First step of major step two. Use numbers to order steps.]

a. [Sub-steps under first step. Use letters for ordering of sub-steps.]
b. [Sub-step two, as needed.]
4. [Second step of major step two] Troubleshooting 1
Step-by-step method example
Step-by-step method details

Live cell analysis: Sir-tubulin labeled microtubule length tracing.

Timing: 2 h (per time-lapse movie)

This step enables the analysis of microtubule dynamics by tracking changes in microtubule
length between successive frames of a time-lapse movie. The analysis accounts for
potential bending or movement of the microtubule and can be performed using tools
available in ImageJ software.

Note: For image preparation, use ImageJ for level adjustments of frame brightness and
contrast, and background subtraction.

1. Microtubule tracing:
a. Use ImageJ to open the movie obtained by recording SiR-tubulin labeled
microglia.
b. Invert LUT using ImageJ dedicated command.
c. …

2. Report the measurements and measure microtubule dynamics parameters:

a. Copy the values into an Excel file.
b. Starting from the second frame, measure the difference in length with the
previous frame.
c. …

Critical: Draw the line as carefully as possible, comparing successive frames through the
entire video to follow the behavior of the entire microtubule. This will help discriminate
the microtubule when the frame is blurred or unclear. See Figure 1 as an example.

Note: As an indication, at least 30 substacks per condition from three independent culture
experiments should be analyzed for a treatment comparison.
 PCR Table Example

PCR reaction master mix

Reagent Amount
DNA template
DNA Polymerase
Primer 1
Primer 2
Buffer x
Buffer y
ddH2O

PCR cycling conditions

Steps Temperature Time Cycles
Initial Denaturation 98 °C 30 sec 1
Denaturation 98 °C 10 sec
Annealing 55 °C 20 sec 25-35 cycles
Extension 72 °C 10 min
Final extension 72 °C X min 1
Hold 4 °C forever

Expected outcomes
EDITOR INSTRUCTIONS
 This section should be written in paragraph form and detailed the expected results from the
protocol and what a researcher can expect to produce for data.
 Include information about anticipated outcomes of the protocol, e.g., estimated yield of DNA
extraction, images of protein expression pattern.
 We encourage authors to provide figures to illustrate the expected outcomes (see examples
here.) and describe the expected results in paragraphs.
 NOTE: If you have no associated primary research manuscript, you are REQUIRED to provide
evidence to validate the data generated by your protocol.
 DO NOT directly insert relevant results sections OR directly reproduce figures from a primary
research paper.
 Find additional instructions for figure preparation in our instructions for authors here.

Quantification and statistical analysis (optional)

EDITOR INSTRUCTIONS
 This section should be written in paragraph form and detail the analysis pipeline.
 Avoid placing extensive stepwise instructions in this section; if the analysis methods require
multiple stepwise instructions, add them as a detailed analysis major step to the protocol.
 Either numbering (up to three levels: 1,2,3; a,b,c; i,ii,iii) or bullet points (up to three levels)
can be used to list steps in this section.
  Provide details about the methods used for data processing, quantification, and statistical
analysis of the data generated in this protocol. Include criteria for data inclusion/exclusion.
 If the protocol outcome requires a complex statistical or computational analysis, include a
sample data set to allow others to repeat your approach, and provide instruction on the
interpretation of the raw data.
 You may provide sample data or reference supplementary data files here.

Limitations
EDITOR INSTRUCTIONS
 In paragraph form, describe possible limitations of the protocol, including any situations
where the protocol may be unreliable or unsuccessful.
 Describe environmental factors or mechanical limitations that might affect the validity of the
results.

Troubleshooting
EDITOR INSTRUCTIONS
 Describe any problems that might arise from running the protocol and provide possible
solutions.
 Make sure to flag them in the corresponding steps of the protocol and refer to this section.
 If applicable, this section can also be the space to include information about alternatives, i.e.,
what reagents and/or equipment has some flexibility and what cannot be changed.
 If you wish to include a list for each problem/potential solution, please use bullet points (up to
three level) make a list.
 DO NOT use numbering (1,2,3; a,b,c; i,ii,iii) in this section.
 We recommend including a minimum of 5 Troubleshooting points.
 Link all Troubleshooting points to the appropriate Steps in the manuscript. Provide the step
number in the Troubleshooting point.

Problem 1:
[When illustrating problems, we encourage the use of figures/videos to indicate a good outcome
versus a bad outcome. See examples here and here.

Potential solution:
[Describe the details of the solution to resolve this problem.]
 Problem 1
Variations in phenotypes due to differences in developmental timing and genetic background
(related to Step 3b).

The use of conditional KD datasets yield variable results even when experiments are
conducted under the same conditions. Additionally, different fly stocks with distinct genetic
backgrounds exhibit variations in their developmental time. These factors contribute to
phenotypic variations, which can complicate experimental analyses and interpretations. To
ensure reliable and accurate outcomes for your experiment, consider implementing the
following potential strategies.

Potential solution
 Phenotypic variation: We acknowledge that some phenotypic variation is inevitable.
However, we have observed that inducing KD earlier in development often results in
stronger phenotypes. Therefore, it is essential to initiate the KD at a similar
developmental stage whenever possible. Minimizing variation in the timing of KD
induction will help reduce phenotypic variability and enhance the reproducibility of
results.
 Egg collection time: The timing of egg collection can significantly affect the
developmental stage of the larvae and, consequently, the resulting phenotypic
outcomes. To minimize variation, ensure precise and consistent egg collection timing.
By reducing the time span for egg collection, we can improve the uniformity of
developmental stages among experimental samples and minimize potential variations
in phenotypes.
 Varying developmental time among different fly stocks: Different fly stocks may exhibit
variations in their developmental timing, which can affect the effectiveness of KD
induction and the resulting phenotypes. To address this, conduct preliminary
experiments to monitor the developmental timing of each specific fly stock used in the
study. This information will allow you to tailor the experimental protocol accordingly,
accounting for the specific differences in developmental stages among different stocks.

Resource availability
EDITOR INSTRUCTIONS
 The lead contact should match the one on the cover page.
 The technical contact should match the one on the cover page.We require four subheadings in
this section (lead contact, technical contact, materials availability, and data and code
availability). The guidelines include instructions and examples.
o Lead contact: Further information and requests for resources and reagents should be
directed to and will be fulfilled by the lead contact, [lead contact’s name] (lead
contact’s email).
 o Technical contact: Technical questions on executing this protocol should be directed
to and will be answered by the technical contact, [technical contact’s name] (technical
contact’s email).
o Materials availability: Provide information regarding availability of newly generated
materials associated with this protocol, including any conditions for access.
o Data and code availability: State whether the protocol includes all datasets
generated or analyzed during this study. Provide information about data and code
availability.

Materials availability statement examples:

 Plasmids generated in this study have been deposited to [Addgene, name
and catalog number or unique identifier].
 Mouse lines generated in this study have been deposited to [the Knockout
Mouse Project (KOMP), name and catalog number or unique identifier].
 This study did not generate new unique reagents.
 There are restrictions to the availability of [reagent] due to [reason why
restrictions exist].

Data and code availability statement examples (Statements with multiple

types of datasets may use a combination of statements)
 Original/source data for [figures/datatype] in the paper is available [e.g.,
Mendeley Data DOI].
 The [datasets/code] generated during this study are available at [name of
repository]: [accession code/web link].
 The published article includes all [datasets/code] generated or analyzed
during this study.
 The [datasets/code] supporting the current study have not been deposited
in a public repository because [reason why data are not public] but are
available from the corresponding author on request.
 This study did not generate/analyze [datasets/code].
There are restrictions to the availability of [dataset/code] due to [reason
why restrictions exist].

Acknowledgments
[Enter the following information here: 1) all funding sources; 2) collaborators and/or core facilities
contributing to the work.]

Author contributions
[Mention each individual author with a statement outlining the contribution of each author to the
work.]
Declaration of interests
[Please disclose competing interests for all submitted content by completing and submitting
the “declaration of interests” form. Include a “declaration of interests” section in the text of
all articles even if there are no interests declared.]

References
EDITOR INSTRUCTIONS
 Please format all in-text citations as superscripted numbers (e.g., “Multiple reports support
this observation.1,2”). All manuscripts must use the superscript numbered Cell Press
referencing style.
 Make sure to use this numbered referencing style for all new and revised submissions as well.
Use the updated CSL and EndNote referencing styles for Cell Press articles.

Figure legends
EDITOR INSTRUCTIONS
 Do not include figures in the main text document. All figures should be uploaded as separate
high-resolution JPG.
 All captions for tables should also be included here.

Figure 1: [Include a clear and concise figure title for each figure. Figure legends are suggested, but not
required.]

Methods Video S1: [Include a clear and concise title for each video. Be sure to mention the relevant
step, e.g., Preparing microscope slides, related to step 3.]

Table 1: [Caption for table with contents.]