Fig 2: VEGFR2 knockdown induces autophagy via the PI3K/AKT/mTOR/ULK1 axis and induces apoptosis by promoting ubiquitination of HSP60. (A) Western blot analysis of PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR, ULK1, p-ULK1, p62 and LC3-II/LC3-I in LX-2 cells transfected with VEGFR2 siRNA and treated with 740Y-P. (B) Colocalization of VEGFR2 and HSP60 as determined by immunofluorescence. (C) Immunoprecipitation analysis of VEGFR2 and HSP60 in LX-2 cells. (D) Western blot analysis of a-SMA, COL1A1, AIF, HSP60, cleaved caspase-9/pro-caspase-9, Bax/Bcl-2 and cytochrome c in LX-2 cells transfected with VEGFR2 siRNA and HSP60 plasmid. (E) JC-1 assay of LX-2 cells transfected with VEGFR2 siRNA and HSP60 plasmid. Data are expressed as the mean ± SD (n=3). (F) Western blot analysis of ubiquitination of HSP60 in LX-2 cells transfected with VEGFR2 siRNA and miR-29b-3p inhibitor. **P<0.01, ***P<0.001. HSP60, heat shock protein 60; miR, microRNA; p-, phosphorylated; siRNA/si, small interfering RNA.