Fig. 2

Reconstitution of PTEN in PTEN-null AN3CA EC cell line. (A) Immunofluorescent detection of PTEN (red) after stable expression of full-length PTEN cDNA in AN3CA, labeled as PTEN-FL. AN3CA cells stably expressing empty pEGFP-C1 control plasmid (green) labeled as Vector. In the overlay panels, yellow squares indicated the areas that were magnified in the zoom panels. Scale bar 50 μm. (B) Immunoblot detected PTEN in PTEN-FL cells after cell fractionation. WCE, SOL, and CHR represented whole-cell extract, soluble, and chromatin fractions, respectively. GAPDH and H3 were used as loading control for soluble and chromatin fractions, respectively. (C) Immunoblot detected phosphorylated Akt at serine 473 residues and total Akt in the whole-cell extracts. GAPDH was used as a loading control. (D) Relative expression of the DDB2 gene was determined by RT-qPCR. The expression values were normalized to an internal control GAPDH. A paired t-test was applied to three independent biological replicates for statistical comparison. (E) Immunoblot detected DDB2 in the whole-cell extracts. GAPDH was used as a loading control