Fig. 4
cZNF215 inactivates the PTEN/AKT pathway by promoting PTEN oxidation. A Western blot assays were conducted to analyze the expression of some relatively crucial proteins of PTEN/AKT pathway in both HuCCT1 and RBE cells. B Representative immunohistochemical (IHC) staining of p-AKTSer473/Thr308 of iCCA tissues with high cZNF215 expression and low cZNF215 expression. C The association between p-AKTSer473/Thr308 and cZNF215 levels of 120 iCCA tissues. D Western blot analysis showing the expression of t-PTEN, p-AKTSer473/Thr308 and t-AKT in HuCCT1 cells transfected with vectors expressing PTEN or in RBE cells transfected with PTEN siRNA. E and F Western blot assays were performed to detect the expression of t-PTEN, p-AKTSer473/Thr308 and t-AKT in HuCCT1 cells cotransfected with indicated vectors and RBE cells cotransfected with indicated siRNAs. G and H Western blot assays were conducted to examine the expression of PTENred, PTENox, t-PTEN, p-AKTSer473/Thr308 and t-AKT in HuCCT1 cells transfected with cZNF215 vectors and treated with NAC, or RBE cells transfected with cZNF215 siRNAs and treated with H2O2. I CCK8 assays showing the proliferation ability of iCCA cells among different groups. J Transwell assays analysis showing the migration and invasion capacity in iCCA cells. Abbreviations: PTENred, reduced PTEN; PTENox, oxidized PTEN; t-PTEN, total PTEN; p-AKTSer473/Thr308, phosphorylation of AKT on Ser473 and Thr308; t-AKT, total AKT; NAC, N-acetylcysteine