Fig. 4. Treatment with miR-142-3p inhibitor increases the levels of autophagy-related protein 16-1 (ATG16L1) messenger RNA (mRNA) and ATG16L1 in in vitro induced Tregs (iTregs) (n = 3).

iTregs were expanded in vitro for 7 days and then treated or not treated with the inhibitor or mimic for 3 days. Western blot and quantitative RT-PCR (qRT-PCR) were used to detect ATG expression and ATG16L1 mRNA levels of iTregs from each group, respectively. a ATG3/5/7/16L1 protein expression determined using western blot. b RNA was purified and qRT-PCR used to determine the expression of ATG16L1 mRNA at different time points (days 3, 6, and 9). c ATG16L1 protein levels of iTregs with or without treatment, measured using western blot. d RNA was purified, and qRT-PCR used to determine the expression of ATG16L1 mRNA of treated and untreated iTregs. e Light chain 3 (LC3) protein expression in iTregs after treatment was detected with western blot. f LC3-II/LC3-I ratios of three groups were analyzed. Results are presented as mean ± standard error of the mean values. *P < 0.05 and **P < 0.01