Figure 2.
PARK2 Depletion Contributes to the Activation of the PI3K/Akt Pathway
(A) Immunoblotting analysis of HCT116 cells stably transfected with control GFP (shGFP) or PARK2 (shPARK2_1 and shPARK2_2) lentiviral hairpins.
(B and C) Shown is the (B) CellTiter 96 AQueous Non-Radioactive Cell Proliferation (MTS) assay and (C) caspase-3/7 activity assay of shGFP or shPARK2 HCT116 cells, following treatment with increasing concentrations of staurosporine (50 and 100 nM) for 1 hr (MTS assay, p = 0.001 for 50-nM and p = 0.0008 for 100-nM treatment; caspase-3/7 assay, p = 0.043 for 50-nM and p = 0.036 for 100-nM treatment).
(D) Immunoblotting analysis of shGFP and shPARK2 HCT116 cells following treatment with the indicated compounds: 1 μM BKM (NVP-BKM120) or 500 nM BEZ (NVP-BEZ235) for 24 hr, 500 nM MK (MK 2206) for 4 hr, 100 nM Rapa (Rapamycin) or Torin for 2 hr, 100 nM PD0 (PD0325901) for 1 hr, or 10 nM GSK (GSK1120212) for 6 hr.
(E) Drug dose-response curves of shGFP and shPARK2 HCT116 cells treated with the indicated compounds for 24 hr (NVP-BKM120, p = 0.014; NVP-BEZ235, p = 9.28 × 10−5; MK 2206, p = 1 × 10−4; Rapamycin, p = 1.45 × 10−11; Torin, p = 6.73 × 10−10; PD0325901, p = 0.02; and GSK1120212, p = 0.03, two-way ANOVA). Data are represented as mean ± SEM (∗p < 0.05 and ∗∗p < 0.01, two-tailed t test).
See also Figure S2.