Fig. 6

cZNF215 facilitates PTEN oxidation by blocking the interaction between PRDX1 and PTEN. A The diagrams of PRDX1 domain structure and Flag-tagged PRDX1 truncations. B Immunoblot assays were performed to detect the expression of PRDX1 truncations and full length of the lysates from HEK293T cells transfected with the indicated vectors (left); RNA pull-down and Immunoblot assays analysis showing the proteins enriched with cZNF215 (right). C RIP assays analysis showing the results from cZNF215 pull-down. D The binding domain of PRDX1 responsible for its interaction with PTEN was validated by Co-IP assays in HEK293T cells. E The interaction between PRDX1 and PTEN was confirmed by Co-IP assays in the HuCCT1 cells transfected with indicated vectors or RBE cells transfected with indicated shRNA. F and G The interaction between PRDX1 and PTEN was further verified by Duolink in situ proximity ligation assay (PLA) assay in the above cells. Scale bars, 50 μm. H The statistical analysis results of average Duolink complexes per cell of PLA assay. I RNA pull-down and immunoblot assays were conducted to detect the cZNF215-PRDX1 complex in HuCCT1 cells transfected with the PTEN siRNA