Fig. 4

SOX2OT promoted autophagy and relieved cell proliferation and fibrosis by inhibiting the Akt/mTOR pathway. Mouse mesangial cells were incubated with high glucose (HG) or normal glucose (NG). HG cells were transfected with pcDNA-SOX2OT or pcDNA-vector. a, f LC3 expression in HG cells after SOX2OT overexpression in the presence or absence of rapamycin was assessed by immunofluorescence. Scale bar: 50 μm. b Western blot analysis of LC3II, LC3I, Beclin1 and p62 in HG cells with SOX2OT overexpression. c, d Expression levels of p-Akt, Akt, p-mTOR, and mTOR in HG cells with SOX2OT overexpression in the presence or absence of rapamycin were determined using western blot assay. e Western blot analysis of LC3II/LC3I, Beclin1 and p62 in HG cells in response to SOX2OT overexpression in the presence or absence of rapamycin. g Western blot assay was implemented to detect expression of TGFβ-1, fibronectin, ASK1 and collagen IV in HG cells with SOX2OT overexpression in the presence or absence of rapamycin. h Expression of E-cadherin and N-cadherin detected by western blot assay in HG cells after SOX2OT overexpression in the presence or absence of rapamycin. GAPDH was used as a loading control in western blot analysis. Error bars represent the mean ± SD from three independent experiments. Rapa, rapamycin. *p < 0.05, **p < 0.01, ***p < 0.001, One-way-ANOVA followed by Newman–Keuls post hoc test