Comprehensive Immune Cell Profiling

The following antibodies were used for staining: CD4 BUV395, CD8α BUV737 (BD); CD5 PerCp-Cy5.5, CD25 PE or APC, CD44 FITC or Pacific Blue, CD45.2 PE, CD62L APC, CD69 APC, TCRβ Pacific Blue, Vα2 PE or Pacific Blue, CXCR5-biotin, PD-1 FITC, SLAM PE-Cy7, CD95 PE-Cy7, GL-7 FITC (Biolegend); B220 efluore780, CD8α efluore780, ZAP70 Alexa488, Strepavidin-A647 (Life Technology); GP66 tetramer PE or APC (NIH Tetramer Core Facility). The following antibodies were used for immunoblot analysis: ZAP70 (clone 1E7.2) (36 (link)); ZAP70-pTyr³¹⁹ (clone 65E4), LAT-pTyr¹⁹¹, Erk1/2, Erk1/2-pThr²⁰²/pTyr²⁰⁴, and PLCγ-pTyr¹⁸³, all from Cell Signaling Technology; anti-phosphotyrosine (4G10), PLCγ (Sigma-Aldrich), LAT (clone FL-233, Santa Cruz Biotechnology), FLAG (clone M2, Sigma-Aldrich), and GAPDH (clone 6C5, Santa Cruz Biotechnology).

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Shen L., Matloubian M., Kadlecek T.A, & Weiss A. (2021). A disease-associated mutation that weakens ZAP70 autoinhibition enhances responses to weak and self ligands. Science signaling, 14(668), eabc4479.

Publication 2021

CD4 BUV395 CD5 PerCp-Cy5.5 CD45.2 PE CD62L APC CD69 APC TCRβ Pacific Blue CXCR5-biotin PD-1 FITC SLAM PE-Cy7 CD95 PE-Cy7 GL-7 FITC Erk1/2 FLAG (clone M2, GAPDH (clone 6C5,

Antibodies Biotin Cd25 Cd44 Cd62l Clone Cxcr5 Cy5 5 Erk1 Fitc Gapdh Immunoblot analysis Phosphotyrosine Plcγ Strepavidin Tcrβ Tetramer Zap70

Corresponding Organization : Howard Hughes Medical Institute

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Variable analysis

independent variables

CD8α
CD45.2
CD62L
TCRβ
CXCR5
CD8α
ZAP70
GP66 tetramer

dependent variables

ZAP70-pTyr319
LAT-pTyr191
Erk1/2
Erk1/2-pThr202/pTyr204
PLCγ-pTyr183
Phosphotyrosine
PLCγ
GAPDH

control variables

Anti-FLAG (clone M2, Sigma-Aldrich)
GAPDH (clone 6C5, Santa Cruz Biotechnology)

Annotations

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