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. Author manuscript; available in PMC: 2012 Apr 1.
Published in final edited form as: Cancer Res. 2011 Feb 11;71(7):2750–2760. doi: 10.1158/0008-5472.CAN-10-2954

Figure 5. PTEN is required for efficient upregulation of BIM following BRAF inhibition.

Figure 5

(A) WM164 cells (PTEN+) were incubated with non-targeting siRNA (NT) or transfected with two PTEN specific siRNA’s (I and II) before treatment with PLX4720 (3 or 10μM, 48hrs). Protein was resolved and probed for expression of BIM, GAPDH and PTEN. (B) Induction of PTEN-wt but not PTEN-G129E in WM793 (PTEN−) cells was sufficient to increase BIM expression when BRAF was inhibited. Left panel: Western blot shows induction of PTEN− wt and PTEN−G129E following doxycycline treatment. Right panel: Induction of PTEN-wt + PLX4720 induces BIM more efficiently than PTEN-G129E + PLX4720. (C) BIM is required for PLX4720-induced apoptosis in PTEN+ cells. WM164 and WM983A cells were incubated with non-targeting siRNA (NT) or transfected with two BIM specific siRNA’s (BIM I and BIM II) prior to treatment with PLX4720 (3 or 10μM, 48 hrs). Flow cytometry studies showed a significant reduction in TMRM loss and Annexin V binding when cells were transfected with BIM siRNA compared to non-targeting control siRNA (*P<0.05).