Fig. 3: KPT-330 reduces Mcl-1 mRNA expression and protein synthesis. | Cell Death & Disease

Fig. 3: KPT-330 reduces Mcl-1 mRNA expression and protein synthesis.

From: XPO1 inhibitor KPT-330 synergizes with Bcl-xL inhibitor to induce cancer cell apoptosis by perturbing rRNA processing and Mcl-1 protein synthesis

Fig. 3

a Real-time PCR analysis of total Mcl-1 mRNA in U87, U251, and H1299 cells treated with KPT-330 (1 μM) for 24 h (mean ± SEM, n = 6 for U87 and U251 and n = 8 for H1299). *P < 0.05. b Real-time PCR analysis of Mcl-1 mRNA decay in U251 and H1299 cells treated with KPT-330 (1 μM) for 24 h and further with Act D (5 μg/ml) for indicated time periods (mean ± SEM, n = 3). *P < 0.05. c The CHX (100 μg/ml) pulse-chase assay in U251 and H1299 cells treated as in a. d, e Western blot analysis of Mcl-1 protein synthesis in U251 and H1299 cells treated with KPT-330 (1 μM) for 24 h, and further with Act D (Act) (5 μg/ml) and MG-132 (MG) (25 μM) for indicated time periods. Quantification of grayscale ratio of Mcl-1/GAPDH by Photoshop software was shown in e (mean ± SEM, n = 3). f Real-time PCR analysis of cytosolic and nuclear Mcl-1 mRNA in U87, U251, and H1299 cells treated as in a (mean ± SEM, n = 3). *P < 0.05. g Western blot analysis of indicated proteins in the cytosol and nucleus of U251 and H1299 cells treated as in a. h Nascent protein synthesis in U87, U251, and H1299 cells treated as in a was measured by OPP incorporation (mean ± SEM, n = 3). *P < 0.05. GAPDH, α-tubulin, and MeCP2 were used as the loading control of total, cytosolic, and nuclear protein, respectively

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