Figure 6
MSC co-culture induces iTreg BACH2 and SENP3 via ROS. (a) Expression of BACH2, SENP3, and FOXP3 on iTregs was measured by immunoblot assay. iTregs were expanded in media vs MSC co-culture and protein was isolated at indicated time. Data are representative of n = 3 independent samples. (b) Analysis of BACH2 and SENP3 on iTregs after CD39 signaling inhibition. iTregs were expanded in media vs MSC co-culture and protein was isolated at indicated time. Expression was measured by immunoblot assay. (c,d) Expression of intracellular reactive oxygen species (ROS) was measured by flow cytometry in media vs MSC co-culture expanded iTregs (c) expression of mitochondrial ROS was measured by FACS analysis (d) at 21 days expansion. Data are representative of n = 6 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 paired t test. See also Supplementary Fig. S7.
