Fig. 5: SVIP Regulates AKT/mTOR Pathway via PTENwt.
From: SVIP reduces IGFBP-2 expression and inhibits glioblastoma progression via stabilizing PTEN
A Co-transfection experiments were conducted in U87-MG cells with PTEN-WT-GFP or PTEN-R130Q-GFP overexpression plasmids, along with empty vectors or SVIP overexpression plasmids. Subsequently, Western blot analysis was employed to assess the protein levels of SVIP, PTEN, and IGFBP-2. B PTEN-WT-GFP and PTEN-R130Q-GFP overexpression plasmids were transfected into U87-MG cells, and the mRNA expression levels of IGFBP-2, LC3B, ATG7, and ATG5 were assessed using qRT-PCR. C PTEN-WT-GFP and PTEN-R130Q-GFP overexpression plasmids were transfected into U87-MG cells, and the protein expression levels and phosphorylation levels of Akt, mTOR, and 4EBP-1 were assessed by Western blot. D The STUB1 overexpression plasmid was transfected into LN229 cells, followed by treatment with or without the LY294002 inhibitor (20 μM, TOPSCIENCE, China). Subsequently, the expression levels of STUB1 and PTEN, as well as the phosphorylation levels of AKT and mTOR, were assessed by Western blot. E The STUB1 overexpression plasmid was transfected into LN229 cells, with or without the LY294002 inhibitor. Subsequently, the expression levels of STUB1, IGFBP-2, and LC3B were assessed by qRT-PCR. F EdU cell proliferation staining was performed to assess the proliferation of U87-MG cells transfected with PTEN-WT-GFP and PTEN-R130Q-GFP overexpression plasmids separately. G Transwell assays were performed to assess the migration of U87-MG cells transfected with PTEN-WT-GFP and PTEN-R130Q-GFP overexpression plasmids individually. Following transfection of PTEN-R130Q-GFP overexpression plasmid into U87-MG cells, transwell assays were conducted to evaluate cell migration after subsequent transfection with STUB1, SVIP, and empty vector. H Immunohistochemical staining was performed for quantitative analysis of SVIP and STUB1 expression in the tissues. Statistical analysis: data were quantified as mean ± SD, n ≥ 3, two tailed student’s t test, P < 0.05, *; P < 0.01, **; P < 0.001, ***; P < 0.0001, ****; ns, no significance.
