Fig. 5

PHB2 promotes SHIP2 protein degradation through ubiquitination. A, B Western blotting (A) and qRT-PCR analysis (B) of SHIP2 expression in HGC-27.sh-PHB2 and MKN-28.sh-PHB2 cells. Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. C Silencing of PHB2 prolonged the half-life time of SHIP2 protein in cycloheximide (CHX)-chase assays. Data are representatives or mean ± SEM; n = 3 independent experiments, two-way ANOVA followed by Šídák’s multiple comparisons test. CHX: 10 μg/ml. D Silencing of PHB2 reduced overexpressed exogenous SHIP2 ubiquitination. MG132:10 μM. Data shown represent three independent experiments. E Overexpression of PHB2 enhanced the ubiquitination of exogenous full-length SHIP2 but not tGFP-SHIP2-del SAM mutant SHIP2. MG132: 10 μM. Data shown represent three independent experiments