Fig. 2

Sinularin induced autophagy through activating ULK1 in prostate cancer cells. (A) Transmission electron micrographs (TEM) showing autophagic vacuoles in PC-3 cells cultured with Sinularin for 24 h. Arrows indicate autophagosome structures. (B) Western blot analysis of the LC3 I/II and p62 levels in LNCaP and PC-3 cells treated with Sinularin for 24 h. (C) Representative immunofluorescence images and mean fluorescence intensity (MFI) quantification of LC3 taken from PC-3 cells treated with Sinularin for 24 h. Autophagosomes were visualized and examined via immunofluorescence microscopy through photographs of LC3 puncta. (D) Western blot analysis was used to determine the phosphorylation levels of mTOR (Ser 2448) and ULK1 (Ser 757) in LNCaP and PC-3 cells treated with Sinularin for 24 h. (E) The cell viability was measured by MTS assay in LNCaP and PC-3 cells treated with Sinularin alone or in combination with ULK1 inhibitor MRT68921. (F) Representative immunofluorescence images and mean fluorescence intensity (MFI) quantification of LC3 taken from PC-3 cells treated with Sinularin alone or in combination with ULK1 inhibitor MRT68921. Autophagosomes were visualized and examined via immunofluorescence microscopy through photographs of LC3 puncta. (G) Western blot analysis of LC3 I/II and p62 levels in LNCaP and PC-3 cells treated with Sinularin alone or in combination with the ULK1 inhibitor MRT68921.