Fig. 5: PDCD11 interacts with p53 in the nucleoplasm.
A IF assay was performed to investigate the subcellular localization of endogenous PDCD11 (red), p53 (green), and their colocalization (yellow) in HCT116p53+/+ cells untreated or treated with Adr (1 µM, 24 h) or Act-D (5 nM, 24 h). The nuclei were stained to blue. Scale bars: 10 µm. B–G GST pulldown assay was performed to investigate PDCD11-p53 interaction in vitro and identify the binding domains. Full-length PDCD11 (PDCD11-FL) and its truncated variants were His-S-tagged, whereas p53-FL and its truncations were GST-tagged. B PDCD11-FL interacts with p53-FL in vitro. C Schematic diagram showing the construction of truncations of p53. D PDCD11-FL interacts with p53 (93-292) and p53 (293-393). E Schematic diagram showing the construction of truncations of PDCD11. F PDCD11 (1-1032) interacts with p53-FL. G PDCD11 (1-1032) interacts with p53 (93-292) and p53 (293-393). H Co-IP assay using anti-p53 (DO-1) was performed to investigate PDCD11-p53 interaction in HCT116p53+/+ cells. Normal mouse IgG was used as a negative control. Input represents 5% of the cell lysate utilized for IP. PLA was performed to investigate the formation (I) and localization (J) of endogenous PDCD11-p53 complex (red) in a panel of CRC cells harboring wild-type p53. The nuclei were stained to blue. Scale bars: 25 µm (I) or 10 µm (J).
