Figure 1.

Gating strategies for identifying T helper 1 (Th1) and 17 (Th17) cells using Kaluza Software. (a) We first identified CD4⁺ T-cells by first identifying singlets (using forward height vs. area scatter), then lymphocytes (using forward vs. side scatter), and then the CD4⁺ lymphocyte population. (b) Th1 cells are CD4⁺CXCR3⁺IL12RB2⁺, so we first gated CD4⁺ and identified CXCR3⁺ cells, which we then gated to identify the IL2RB2⁺ cells. We used negative isotype control at 1% to distinguish positive populations from non-specific background signals. (c) Th17 cells are CD4⁺CCR6⁺IL23R⁺, so we first gated CD4⁺ and identified CCR6⁺ cells, which we then gated to identify the IL23R⁺ cells. We used negative isotype control at 1% to distinguish positive populations from non-specific background signals.