Fig. 8
Ouabain modulates key apoptosis regulatory proteins in both intrinsic and extrinsic pathways of apoptosis in AML cells. A, B AML KG-1 cells were treated with ouabain (0 − 40 nM) for 48 h. A Western blot analysis of the key apoptosis regulatory proteins, including Bcl-2, Bax, Bid, Mcl-1, c-Myc, c-FLIP, and Fas. β-actin was used as a loading control. Densitometric analysis of Bcl-2 and Bax presented as a ratio of Bcl-2 to Bax, Mcl-1, c-Myc, c-FLIPL, c-FLIPS, and Fas levels is shown. Data are mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 versus nontreated control cells; two-sided Student's t test. B qPCR of MCL1, MYC, CFLAR (encoding c-FLIP), and FAS mRNA expression. GAPDH was used as a house-keeping gene. Data are mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 versus nontreated control cells; two-sided Student's t test. C, D Western blot analysis of Mcl-1, c-Myc, c-FLIPL, and c-FLIPS in enriched LSCs and LPCs from AML KG-1 (C) and Kasumi-3 (D) cells. Data are mean ± SD (n = 4). *P < 0.05, **P < 0.01 versus enriched LSCs; two-sided Student's t test