Nature - USA (2020-09-24)

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nature research | reporting summary

October 2018

Field-specific reporting

Please select the one below that is the best fit for your research. If you are not sure, read the appropriate sections before making your selection.

Life sciences Behavioural & social sciences Ecological, evolutionary & environmental sciences

For a reference copy of the document with all sections, see nature.com/documents/nr-reporting-summary-flat.pdf

Life sciences study design

All studies must disclose on these points even when the disclosure is negative.

Sample size No statistical methods were used to predetermine sample sizes. Sample sizes were determined based on previous studies from our group and

publications in the field.

Data exclusions No data were excluded from the analysis

Replication Experiments were replicated several times with reproducible results, as indicated in each figure legend.

Randomization Mice were paired based on gender (male), weight and age, and randomly selected for brain ischemia experiments.

Blinding The data collection was not blinded. Blinding was not possible as the investigators were also conducting the experiments and had to be aware

of controls and treated groups.

Reporting for specific materials, systems and methods

We require information from authors about some types of materials, experimental systems and methods used in many studies. Here, indicate whether each material,

system or method listed is relevant to your study. If you are not sure if a list item applies to your research, read the appropriate section before selecting a response.

Materials & experimental systems

n/a Involved in the study

Antibodies

Eukaryotic cell lines

Palaeontology

Animals and other organisms

Human research participants

Clinical data

Methods

n/a Involved in the study

ChIP-seq

Flow cytometry

MRI-based neuroimaging

Antibodies

Antibodies used For immunohistochemistry, the following antibodies were used in this study. anti-mouse Tubulin b3 (no.801208, Biolegend,

Alexa Fluor 594 conjugate, clone TUJ1), anti-mouse c-Fos (no.SC-52, Santa Cruz Biotechnology), anti-mouse pERK (no.9101, Cell

Signaling Technology), anti-mouse PGP9.5 (no.ab8191, Abcam, clone 13C4/I34), anti-mouse TRPV1 (no. GP14100, Neuromics),

anti-mouse Tubulin b3 (no.ab68193, Abcam, clone EPR1568Y), anti-mouse Tyrosine hydroxylase (no. AB152, Merck Millipore),

anti-mouse HuC/D (no. A-21271, Thermo Fisher Scientific, clone 16A11), anti- rabbit IgG antibody (no.ab150077, Abcam, Alexa

Fluor 488 conjugate), anti- mouse IgG antibody (no.ab175473, Abcam, Alexa Fluor 568 conjugate), anti- guinea pig IgG antibody

(no.ab150187, Abcam, Alexa Fluor 647 conjugate), anti- mouse Tubulin b3 (no.801203, Biolegend, Alexa Fluor 488 conjugate,

clone TUJ1), anti- mouse I-A/I-E (no.107617, Biolegend, Alexa Fluor 647 conjugate, clone M5/114.15.2).

For flow cytometry analysis, the following antibodies were used in this study. anti-mouse MHC-II (no.12-5321-82, eBioscience, PE

conjugate, clone M5/114. 15.2), anti-mouse TCRb (no.109208, Biolegend, PE conjugate, clone H57-597), anti-mouse CD3e

(no.552774, BD Biosciences, PE-Cy7 conjugate, clone 145-2C11), anti-mouse CD3e (no.561824, BD Biosciences, PE conjugate,

clone 145-2C11), anti-mouse B220 (no.103208, Biolegend, PE conjugate, clone RA-6B2), anti-mouse NK1.1 (no.108708,

Biolegend, PE conjugate, clone PK136), anti-mouse CD11c (no.558079, BD Biosciences, PE-Cy7 conjugate, clone 1HL3), anti-

mouse Foxp3 (no.45-5773-80, eBioscience, PerCP conjugate, clone FJK-16s), anti-mouse CD4 (no.563106, BD Biosciences, BV510

conjugate, clone RM4-5), anti-mouse RORgt (no.562894, eBioscience, Alexa Fluor 488 conjugate, clone Q31-378), anti-mouse

Helios (no.137216, Biolegend, PE conjugate, clone 22F-6), anti-mouse IFNg (no.505808, Biolegend, PE conjugate, clone XMG1.2),

anti-mouse Gata3 (no.53-9966-42, eBioscience, Alexa Fluor 488 conjugate, clone TWAJ), anti-mouse IL-17A (no.45-7177-82,

eBioscience, PerCP conjugate, clone 17B7), anti-mouse NKp46 (no.11-3351-82, eBioscience, Alexa Fluor 488 conjugate, clone

11-3351-82), anti-mouse IL-22 (no.17-7222-82, eBioscience, APC conjugate, clone IL22JOP), anti-mouse CD24 (no.138504,

Biolegend, PE conjugate, clone 30F-1), anti-mouse CD184 (no.146508, Biolegend, APC conjugate, clone L276F12), anti-mouse

CD44 (no.103030, Biolegend, PE-Cy7 conjugate, clone IM7), anti-mouse CD16/CD32 (no.553141, BD Bioscience, clone 2.4G),

anti-human CD45 (no.304038, Biolegend, FITC conjugate, clone HI30), anti-human CD3 (no.47-0036-41, eBioscience, APC-eFluor

780 conjugate, clone SK7), anti-human CD19 (no.47-0199-41, eBioscience, APC-eFluor 780 conjugate, clone HIB19), anti-human

CD56 (no.318331, Biolegend, APC-Cy7 conjugate, clone NCAM), anti-human HLA-DR (no.307646, Biolegend, BV510 conjugate,