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Figure 4 | Molecular Cancer

Figure 4

From: The Bcl-2/xL inhibitor ABT-263 increases the stability of Mcl-1 mRNA and protein in hepatocellular carcinoma cells

Figure 4

ABT-263 increases the protein stability of Mcl-1. (A, B) After pretreated with 5 μM ABT-263 or vehicle DMSO for 12 h, PLC cells were treated with 10 μM CHX for indicated times. The level of Mcl-1 protein was detected by Western blot taking α-tubulin as a loading control (A), and the relative level of Mcl-1 protein was calculated (B) as follows: (1) quantifying the mean ratio of Mcl-1/α-tubulin for each sample according to the Western blot results from triplicate assays and (2) calculating the fold induction of Mcl-1 by normalizing the mean Mcl-1/α-tubulin ratio of the tested sample to that of corresponding control (CHX 0 min). Data were expressed as mean ± SD from three independent experiments. (C) After pretreated with 10 μM proteasome inhibitor MG132 for 2 h, the HCC cells were incubated with 5 μM ABT-263 or vehicle DMSO for 6 h. Then the level of Mcl-1 protein was detected by Western blot. (D) HCC cells were transfected with USP9X-specific siRNA or control siRNA for 36 h, then the mRNA level of USP9X was analyzed by RT-PCR. (E) After transfected with USP9X-specific siRNA or control siRNA for 24 h, the HCC cells were treated with 5 μM ABT-263 or vehicle DMSO for another 18 h. Then the level of Mcl-1 protein was analyzed by Western blot.

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