Flotillin-1 promotes the progression of hepatocellular carcinoma by activating TFE3-mediated Golgi stress response via inhibition of mTORC1/2

Figure 3 Flotillin-1 promotes Golgi stress in hepatocellular carcinoma. A and B: Flotillin-1 overexpression was transfected in Huh-7 cells and flotillin-1 short-hairpin RNA was transfected in SNU-387 cells with H₂O₂ (0, 100, 200, 500, and 1000 μM) stimulation. CCK-8 assay was used to detect cell viability of Huh-7 (A) and SNU-387 (B) cells; C-E: Huh-7 cells and SNU-387 cells were stimulated with H₂O₂ (1000 μM). Fragmentation of Golgi apparatus was observed by immunofluorescence staining with Golgi marker protein GM130. Scale bar = 50 μm (C). Expression of Golgi stress-related proteins (GM130, giantin, p115, and Golgi phosphoprotein 3) in Huh-7 and SNU-387 cells was detected by western blot (D and E). Values are expressed as mean ± SD (n = 3), and comparisons between two groups were performed using the t-test; comparisons among multiple groups, one-way ANOVA was employed, followed by the LSD test or Tamhane’s T2 test. ^aP < 0.05, ^bP < 0.01. FLOT1: Flotillin-1; OV: Overexpression; DAPI: 4’-6-diamidino-2-phenylindole; GM130: Golgi matrix protein 130; shRNA: Short-hairpin RNA; GOLPH3: Golgi phosphoprotein 3.