Extended Data Fig. 8: Gating strategy for flow cytometry analysis.
From: THEMIS is a substrate and allosteric activator of SHP1, playing dual roles during T cell development
a, Gating strategy for T cell development analysis in total thymocytes. Cells were first gated by FSC/SSC to exclude debris, followed by gating FCS-A/FCS-W and SSC-A/SSC-W to eliminate non-single cells. Live cells were gated by Indo-1(Violet) negative, followed by total thymocyte development analysis. b, Gating strategy for T cell development analysis in total splenocytes. Cells were first gated by FSC/SSC to exclude debris, followed by gating FCS-A/FCS-W and SSC-A/SSC-W to eliminate non-single cells. Live cells were gated by Pacific Blue negative, followed by total splenocytes development analysis. c, Gating strategy for THEMIS protein expression in DP thymocytes. Cells were first gated by FSC/SSC to exclude debris, followed by gating SSC-A/SSC-W and FCS-A/FCS-W to eliminate non-single cells. Different mice cells were distributed by distinct FITC/BV421 staining, and then DP cells were gated by CD4-BV605 and CD8-APC-Cy7 double positive population. THEMIS protein expression was stained by APC.
