Fig. 6: PDCD11 interacts with HDM2 in the nucleoplasm. | Oncogenesis

Fig. 6: PDCD11 interacts with HDM2 in the nucleoplasm.

From: Programmed cell death 11 modulates but not entirely relies on p53-HDM2 loop to facilitate G2/M transition in colorectal cancer cells

Fig. 6

A IF assay was performed to investigate the subcellular localization of endogenous PDCD11 (red), HDM2 (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–F GST pulldown assay was performed to investigate PDCD11-HDM2 interaction in vitro and identify the binding domains. PDCD11-FL and its truncations were His-S-tagged, whereas HDM2-FL and its truncations were GST-tagged. B PDCD11-FL interacts with HDM2-FL in vitro. C Schematic diagram showing the construction of truncations of HDM2. D PDCD11-FL interacts with HDM2 (349-497). E PDCD11 (1033-1431) interacts with HDM2-FL. F PDCD11 (1033-1431) interacts with HDM2 (349-497). G Co-IP assay using anti-HDM2 (D1V2Z) was performed to investigate PDCD11-HDM2 interaction in HCT116p53+/+ cells. Normal rabbit IgG was used as a negative control. Input represents 5% of the cell lysate utilized for IP. PLA was performed to investigate the formation (H) and localization (I) of endogenous PDCD11-HDM2 interaction (red) in a panel of CRC cells harboring wild-type p53. The nuclei were stained to blue. Scale bars: 25 µm (H) or 10 µm (I).

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