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. 2022 Jul 8;23(9):e54677. doi: 10.15252/embr.202254677

Figure EV3. Flow cytometry gating strategy for Fig 5 .

Figure EV3

  • A
    Exemplary electronic gating strategy for Fig 5B and C. OTII T cells were identified based on their co‐expression of TCR Vα2 and β5 and Tfh cells by co‐expression of PD‐1 and CXCR5.
  • B
    Gating strategy for Fig 5D–G. Gates 1–4 were activated in SpectroFlo software after which CD45.2‐positive cells were exported for analysis using FlowJo.
  • C
    Proliferation analysis by CTV dilution. WT or Il21r −/− SWHEL B cells from individual mice were concatenated into one file per experiment. CTV division peaks were then identified using FlowJo's Proliferation tool by first identifying peaks 0–7 and then 8–11+. Gray bars indicate gate used to set the initial peak (0 or 4). For statistical analysis in Fig 5E, the resultant electronic gates for each CTV division were then applied to individual mice.