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. 2021 Dec 15;131(24):e140436. doi: 10.1172/JCI140436

Figure 3. WWP1 expression is upregulated in isogenic breast cancer cells resistant to PI3K inhibitors.

Figure 3

(A) Evaluation of WWP1 and Myc expression, and activity of the downstream PI3K pathway in isogenic resistant MDA-MB-231 and MCF7 cells. A set of isogenic resistant cells was sequentially established by repeating exposure to stepwise increasing concentrations of BKM120. Generated clones were maintained in BKM120-containing media. For the experiment, cells were cultured with BKM120-free media for 24 hours and harvested for WB. (B) Endogenous PTEN ubiquitination in isogenic MDA-MB-231 cells. Extracted lysates were pulled down by anti-PTEN antibody. (C) Activation of AKT and S6 in MDA-MB-231 cells with stable overexpression of WWP1 upon BKM120 treatment. Total lysates were resolved by SDS-PAGE and subsequently probed with the indicated antibodies. EV, control empty vector. (D and E) Dose response curve of BKM120 in MDA-MB-231 (D) and BYL719 in MCF7 (E) cells with stable overexpression of HA-WWP1. (F and G) Dose response curve of BKM120 in resistant MDA-MB-231 HD cells (F) and BYL719 in resistant MCF7 HD cells (G) with WWP1 shRNA. Each dot indicates the mean ± SD of a duplicated assay.