Additional file 4: of Oncogenic Ras is downregulated by ARHI and induces autophagy by Ras/AKT/mTOR pathway in glioblastoma

ARHI over-expression can induce autophagy and inhibit proliferation in patient-derived primary cells. (a) Fluorescence microscopy of LC3B expression after transfection with ARHI and negative control plasmids. The scale bar represents 100 μm. (G#2: patient2-derived cells) (b) After transfecting with ARHI and negative control plasmids for 72 h, the expression levels of ARHI, SQSTM1,LC3B,Ras,phosphorylated and total mTOR and AKT were assessed by western blotting. Using total mTOR and AKT as the internal controls, the expression levels of phosphorylated mTOR and AKT were calculated (G#3: patient3-derived cells, the bar whiskers represent SD, *p < 0.05, **p < 0.01, ***p < 0.001). (c) Cell viability of patient3-derived cells after over-expressing ARHI or inhibiting autophagy by shATG5 (G#3: patient3-derived cells,the bar whiskers represent SD, *p < 0.05, **p < 0.01, ***p < 0.001). (d) The intracranial tumor size in orthotopic xenograft model. Scale bar represent 2000 μm. The survival time of nude mice bearing glioma. (G#1: patient1-derived cells) (JPG 1224 kb