TFEB and TFE3 are necessary for SB202190-induced autophagy and lysosomal biogenesis. (A) SB202190 increased the expression of multiple autophagy-lysosomal pathway related genes. After HeLa cells were treated with SB202190 (10 μM) for 12 h, the expression of several autophagy- and lysosome-related genes were detected by real-time PCR. (B) Quantification data of TFEB and TFE3 levels after transfection of HeLa cells with TFEB and TFE3 siRNAs for 48 h. (C) Knockdown of the expression of Tfeb and Tfe3 attenuated SB202190-induced expression of several autophagy and lysosome-related genes as reflected by real-time PCR assay. (D) Tfeb or/and Tfe3 knockdown attenuated SB202190-induced expression of LAMP1, SQSTM1/p62 and LC3B-II levels in HeLa cells. HeLa cells were treated with SB202190 (10 μM) for 16 h after transfection of cells with indicated SiRNAs for 48 h. The expressions of LAMP1, SQSTM1/p62, MAP1LC3, TFEB, and TFE3 were detected. (E) Quantification data showed that depletion of TFEB/TFE3 partially but significantly attenuated SB202190-induced expression of LC3B-II levels. Quantitative data are presented as the mean ± SEM from at least 3 independent experiments.