Figure S5.
The flow cytometry plots on the left and data summary bar graphs on the right. The flow cytometry plots display the levels of phosphorylated STAT6 (pSTAT6) and phosphorylated STAT5 (pSTAT5) for IL-4, and phosphorylated STAT3 (pSTAT3) and phosphorylated STAT5 (pSTAT5) for IL-21, in response to different concentrations of IL-4 or IL-21. The bar graphs summarize the MFI of phosphorylated STAT proteins, comparing untreated, 2.5 nanograms per milliliter IL-4, 10 nanograms per milliliter IL-4, and 100 nanograms per milliliter IL-4 for the IL-4 response, and untreated, 100 nanograms per milliliter IL-21, and 500 nanograms per milliliter IL-21 for the IL-21 response. The axes of the bar graphs are labeled with the MFI values on the y-axis and the treatment conditions on the x-axis. The data show significant differences in pSTAT levels between different treatment conditions, as indicated by the p-values. The flow cytometry plots and bar graphs together illustrate how 3A10t Tregs respond to varying concentrations of IL-4 and IL-21 by altering the phosphorylation levels of specific STAT proteins.
Response of 3A10t Tregs to IL-4 and IL-21 (related to Fig. 6). 3A10t- and EV-transduced Tregs were washed and cultured in IL-2–free media overnight. The cells were then stimulated with the indicated concentrations of IL-4 (upper panels) or IL-21 (lower panels) for 30 min, followed by flow cytometric analysis for pSTAT3, pSTAT6, and pSTAT5. Representative flow cytometry plots are shown on the left and data summary are shown on the right. Statistical significance of the difference was determined using two-way ANOVA followed by Tukey’s multiple comparison test.