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. Author manuscript; available in PMC: 2018 Dec 21.
Published in final edited form as: Mol Cell. 2017 Dec 7;68(6):1134–1146.e6. doi: 10.1016/j.molcel.2017.11.006

Figure 3. Nuclear localization of phosphorylated p53-RS is dependent on Pin1.

Figure 3.

A) Time-lapse analysis of GFP-p53 in H1299. GFP-p53-RS was stably expressed in H1299 cells. Frames show the GFP-p53-RS subcellular localization; Arrows indicate the one cell division in mitosis to two daughter cells in G1 phase. For each sequence, t=0 min is the first frame marked by the arrow were observed. Scale bar, 10um. The same for panels B and C below.

B) Time-lapse analysis of GFP-p53 after treatment of 500 nM CDK4 inhibitor (CDK4i) in H1299 cells.

C) Time-lapse analysis of GFP-p53 after treatment with 5 μM Pin1 inhibitor (Pin1i) in H1299 cells.

D) Knockdown of Pin1 decreases nuclear p53-RS level. PLC/PRF/5 cells were transfected with siRNA for Pin1 (SiPin1) or p53-RS (Sip53), and used for subcellular fractionation 72 h after transfection, followed by WB with indicated antibodies.

E) Nuclear translocation of phosphorylated p53-RS is blocked by a transport inhibitor Importazole. H1299 cells were transfected with combination of plasmids encoding p53-4MRS, and/or Flag-PIN1 and treated with Importazole overnight before harvesting for analysis of subcellular fractions by co-IP with IgG or a p53 antibody followed by WB with indicated antibodies.