(A) Real-time qRT-PCR analysis of PDHK1 mRNA expression in PTEN-deficient A2058 cancer cells stably expressing PTENWT or PTENG129E or PTENY138L or GFP. Data are shown as mean ± SD (n = 2 replicates). ****p < 0.0001; n.s., not significant compared with “GFP-expressing PTEN-deficient cells” by Tukey’s multiple-comparisons one-way ANOVA.
(B) Western blots showing phospho-AKT and PDHK1 expression in PTEN-deficient cancer cell lines in response to 1 μM BKM-120 (PI3-kinase inhibitor) or vehicle (DMSO) treatment for 24 h.
(C) Western blots showing phospho-AKT, phospho-S6(mTOR effector), and PDHK1 expression in PTEN-proficient cancer cell lines expressing empty vector (EV) or myristoylated-AKT (Myr-AKT) to constitutively activate AKT signaling.
(D) Effects of pharmacologic inhibition of PDHK1 with DCA (dose response: 5, 10, and 20 mM) in PTEN-deficient cancer cell lines stably expressing PTENW7 or PTENG129E or PTENY138L or GFP on cell growth by crystal violet staining assay are shown, with quantification of cell viability in 20 mM DCA treatment relative to vehicle (water) treatment reported as rescue score (STAR Methods).
See also Figure S7.