Figure 4.

OC fails to increase TFEB-luciferase activity and lysosome biogenesis. a HeLa cells were treated with OC (15 μM) for a certain time period and analyzed by qRT-PCR for TFEB target gene expression. b 293T cells were transiently transfected with a TFEB-luciferase reporter vector together with a Renilla luciferase vector for 24 h. Cells were then treated with OC (15 µM) for a certain time period or Torin1 (1 μM) for 4 h, and the luciferase activity was evaluated. *P < 0.05, treatment with OC and Torin1 compared to DMSO. (c) HeLa cells were treated with OC (15 μM) or PP242 (1 μM) for 8 h, labeled with LysoTracker Red (50 nM) and analyzed by immunofluorescence and flow cytometry. d HeLa cells were treated with OC (15 μM) or PP242 (1 μM) for 8 h. Cells were labeled with Magic Red Cathepsin B or L reagent for 30 min and measured by flow cytometry