Fig 2: gRb1 increases STAT5 phosphorylation and NFAT1 activation in Tregs. Splenocytes from naïve mice were stimulated with rhIL‐2 in the presence of gRb1 or vehicle control for 15 min. (A) Live lymphocytes were selected based on FSC and SSC (Plot #1). CD4+ T cells were identified by CD4 staining (Plot #2). Finally, Tregs and Tconvs were identified by Foxp3 expression in CD4+ T cells (Plot #3). (B and C) Frequencies of pSTAT5+ cells among Tregs (blue gate) (B) and Tconvs (red gate) (C) stimulated with rhIL‐2 (1–100 ng/mL) in the presence of gRb1 (100 μM) or vehicle. The percentages of pSTAT5+ cells are shown in the line graphs (n = 4 mice per group). One‐way ANOVA with Tukey's test. Bars indicate the mean ± SEM. (D) Representative plots illustrating frequencies of pSTAT5+ cells among Tregs (blue gate) and Tconvs (red gate) stimulated with rhIL‐2 (20 ng/mL) in the presence of gRb1 (50–200 μM) or vehicle. One‐way ANOVA with Tukey's test. (E) Plots illustrating NFAT1+Foxp3+ (NFAT1+ Tregs) and NFAT1+Foxp3− (NFAT1+ Tconvs) proportions among CD4+ cells. The box‐and‐whisker plots present the percentages of NFAT1+ Tregs (left) and NFAT1+ Tconvs (right). Mann–Whitney U test, *p < 0.05, **p < 0.01, ****p < 0.0001. The data are representative of two separate experiments. The box in a box‐and‐whisker plot extends from the 25th to the 75th percentile, and the whiskers are plotted from the minimum to the maximum. It also plots each individual value as a point overlaid on the graph; n = 3–5 animals in each group. gRb1, ginsenoside Rb1; SSC, Side Scatter; FSC, Forward Scatter; CD, Cluster of Differentiation; Foxp3, Forkhead Box P3; pSTAT5, phospho‐Signal Transducer and Activator of Transcription 5; IL, Interleukin; Tregs, Regulatory T Cells; Tconvs, Conventional T cells; rhIL‐2, recombinant human IL‐2; NFAT1, Nuclear Factor of Activated T‐cells 1.