Fig. 3.

Phosphorylation of TRIM59 at Ser308 recruits PIN1. a IP and WB for TRIM59 binding to PIN1 in LN229/EGFR and U87/EGFR cells with or without EGF stimulation. b IP and WB for TRIM59 association with PIN1 in LN229/EGFR cells stimulated with EGF (100 ng/ml) for 6 h. Cells were pre-treated with or without Roscovitine (20 μM) for 1 h. c Effects of ectopic expression of TRIM59 WT, S308A, or S308D mutant on TRIM59 association with PIN1 with or without EGF stimulation. d Effects of ectopic expression of PIN1 WT or WW mutant on TRIM59 binding with PIN1. e Cis–trans isomerization analysis. The synthesized phosphorylated or nonphosphorylated oligopeptides of TRIM59 were mixed with purified PIN1 WT or C113A mutant. Data were expressed as means ± SD. P values were calculated using one-way analysis of variance (ANOVA). f WB for effect of PIN1 knockout with a PIN1 sgRNA on TRIM59 nuclear translocation in LN229/EGFR GBM cells. g Effects of re-expression of shRNA-resistant PIN1 WT or C113A mutant on TRIM59 nuclear translocation. LN229/EGFR GBM cells reconstituted with empty vector (EV), shRNA-resistant PIN1 WT, or C113A mutant were stimulated with or without EGF (100 ng/ml) for 6 h. Data are representative of three independent experiments with similar results. Source data are provided as a Source Data file