Fig. 3

IMP3 promoted the activation of PI3K/AKT/mTOR signaling and facilitated the ubiquitination of PTEN protein in prostate cancer cells. a-c. The expression levels of PIP2, PIP3, p-AKT, AKT, p-mTOR, mTOR, PTEN and BAD were determined by western blotting technology after IMP3 were upregulated or downregulated in PC3, LNCap and DU145 cells. d-f. Following cell transfection/infection with si-IMP3, si-NC, OE-IMP3 or OE-NC for 24 h, DU145 and LNCap cells were treated with 100 μg/ml of CHX for 0, 1, 2, 4, 8 or 24 h, then total proteins were extracted from cells and submitted to western blotting to detect the expression of PTEN protein. g-i. The ubiquitination level of PTEN protein was determined by IP assay after cell transfection/infection with si-IMP3, si-NC, OE-IMP3 or OE-NC in DU145 and LNCap cells with anti-PTEN antibody. j-l. The ubiquitination level of PTEN protein was determined by IP assay with anti-Ub antibody. m-n. The ubiquitination level of PTEN protein was determined by IP assay after cell treatment with MG132 (10 μg/ml) for 4 h or without. (^*p < 0.05, ^**p < 0.01, si-IMP3 group compared with si-NC group; ^#p < 0.05, ^##p < 0.01, OE-IMP3 group compared with OE-NC group)