Fig. 4

CCR6 suppresses TFEB nuclear translocation to block autophagic flux by binding to TFEB. A The protein level of TFEB in the nucleus and cytoplasm of ESCs with different treatments by western blot. B Quantification of the gray value of TFEB western blot bands in the nucleus and cytoplasm of ESCs. C Representative immunofluorescence images of subcellular localization of TFEB in different treated ESCs. (original magnification 1000 ×) D Quantitative fluorescence intensity of subcellular localization of TFEB. E Representative immunofluorescence images of Gal3 in different treated ESCs. (original magnification 200 ×) F Quantitative fluorescence intensity of Gals. G Representative immunofluorescence images of Lyso-Tracker Green in different treated ESCs. (original magnification 200 ×) H Quantitative fluorescence intensity of Lyso-Tracker. I Quantification of acid phosphatase activity in different treated ESCs. J Representative confocal images of ESCs with different treatments were transfected with Ad‐LC3 in the presence or absence of 100 nM Baf-A1 treatment. (original magnification 1000 ×) K Quantification of LC3 puncta number of representative cells. L Binding relationship between CCR6 and cytosolic TFEB by co-IP experiment. Data are presented as the mean ± SD of n = 3 independent experiments. Statistical analysis was performed using one-way ANOVA with Tukey post hoc (B, D, K) or Mann–Whitney test (F, H, I). ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns, non-significant