Fig. 3
Genistein dose-dependently impaired Treg function. (A) RT-qPCR analysis of Treg functional markers (Foxp3, CTLA-4, and LAG-3) after 24-h treatment with genistein at concentrations of 0–20 µM. (B) Western blot detection of Foxp3 protein expression following 24-h exposure to genistein (0–20 µM). (C) ELISA quantification of IL-10 in Treg culture supernatants after 24-hour genistein treatment (0–20 µM). (D) CD4+CD25− Th cells (2 × 105/ml) were labeled with CFSE and subsequently cocultured with Tregs preincubated with genistein (10 µM) for 48 h. The Th cell proliferation was determined by flow cytometry. (E) ELISA quantification of IFN-γ and TNF-α in supernatants from Treg and Th cell co-cultures. (n = 3). Data are showed as mean ± SD; *p < 0.05; **p < 0.01; ***p < 0.001.
