Fig. 5

The inhibitor of hVps34 can block SGK3 activity and suppress liver CSC expansion after prolonged treatment of HCC cells with PI3K inhibitors. a Huh7 and MHCC97H cells were treated with VPS34-IN1 for 24 h, and cell lysates were subjected to western blot analysis with the indicated antibodies. b Spheroid formation assay of MHCC97H cells treated with VPS34-IN1 (top), the statistical results of the tumour spheroids (> 50 μm) were calculated based on 3 independent experiments (bottom). Scale bars, 200 μm. c Immunofluorescence images of Nanog (red) in MHCC7H cells after treatment with VPS34-IN1 or dimethyl sulphoxide DMSO for 24 h. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. d The proportion of CD133+ cells in Huh7 cells was evaluated by flow cytometric assay. e Treatment of MHCC97H cells with VPS34-IN1 for 24 h inhibited the overexpression of CD133 and Nanog induced by treatment with ZSTK474, detected by RT-PCR. *P < 0.05