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X. Li et al.
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Figure 5. CUDC-907 synergizes with venetoclax by targeting Mcl-1 protein stability and transcriptionally regulating Bim, CHK1, Wee1, and RRM1. (A) Acute myeloid leukemia AML cells were treated for 24 h with vehicle control, venetoclax (VEN), CUDC-907 (CUDC), or in combination at the indicated concentrations. Total RNA was isolated and then Mcl-1, Bim, Wee1, CHK1, and RRM1 transcripts were determined by real-time RT-PCR. *P<0.05, **P<0.01, and ***P<0.001 compared to vehicle control. (B, C) MOLM-13 (panel B) and U937 (panel C) cells were treated with vehicle control, venetoclax, CUDC-907, or in combination for 12 hours, washed and then treated with 10 mg/mL cycloheximide for up to 2 hours. Western blots were probed with anti-Mcl-1 or -b-actin antibody. The fold changes for the densitometry measurements of Mcl-1 were normalized to β-actin and then compared to vehicle control. Representative western blots are shown on the left, while densitometry measurements are graphed and shown on the right. ns indicates not significant, *P<0.05, **P<0.01, and ***P<0.001. (D-F) MOLM-13 (panel D), U937 (panel E), and patient sample AML#23 (panel F) cells were treated with vehicle control, venetoclax, CUDC-907, or in combination with or without MG132, at the indicated con- centrations, for 24 hours. Western blot analysis of whole cell lysates are shown. The fold changes for the densitometry measurements, normalized to b-actin and then compared to vehicle control, are shown.
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