Fig. 3
From: WWP1 targeting PTEN for polyubiquitination to promote bone metastasis of luminal breast cancer
WWP1 polyubiquitylates PTEN. (A) WWP1 was knocked down in MCF7 cells using shRNA, and knockdown efficiency was verified through western blotting. (B) Quantification of WWP1 (left) and PTEN (right) protein levels normalized to GAPDH in (A) is shown. (C) Effects of WWP1 knockdown on WWP1-mediated PTEN polyubiquitination. WWP1-silenced MCF7 cells were transfected with the indicated constructs, and PTEN polyubiquitination was analyzed through western blotting. (D) Quantification of the ubiquitination levels of PTEN in (C) is shown. (E) Analysis of PTEN polyubiquitination level in WWP1-knockdown MCF7 cells with ectopic overexpression of either wide-type WWP1 (WT) or C890A WWP1 (CA, a catalytically inactive mutant of WWP1). (F) Quantification of the ubiquitination levels of PTEN in (F) is shown. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, results are representative of at least three independent experiments and shown as mean ± SD.
