Figure 3.

OC-induced TFEB nuclear translocation is dependent on mTORC1 activity. a OC attenuated the interaction of TFEB and 14-3-3 proteins. 293 T cells were transfected with a Flag-TFEB plasmid and the next day treated with OC (15 μM) or PP242 (1 μM) for 8 h. Cells were lysed and Flag-TFEB was immunoprecipitated with anti-Flag beads. Cell lysates (input) and immunoprecipitated proteins were immunoblotted with anti-14-3-3, anti-Flag and anti-GAPDH antibodies. Relative 14-3-3/Flag (IP) ratios were measured using ImageJ software. ^*P < 0.05, treatment with OC and PP242 compared to DMSO. b, c HeLa or MEF cells were treated with OC (15 μM) or Torin1 (1 μM) for a certain time period. TFEB, P-p70S6K and P-S6 were analyzed by Western blotting with the indicated antibodies. ^*P < 0.05, treatment with OC compared to DMSO. d TSC2^+/+ or TSC2^-/- MEF cells were treated with OC (15 μM) or PP242 (1 μM) for a certain time period. P-p70S6K and P-S6 were analyzed by Western blotting