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. 2021 Dec 8;60(1):2. doi: 10.3892/ijo.2021.5292

Figure 4.

Figure 4

MCL-1 downregulation occurs at transcriptional and post-translational levels. (A) MDA-MB-231 and HS578T cells were treated using 5 µM ACF and the expression of MCL-1 mRNA was evaluated after 0, 1, 3, and 6 h using reverse transcription-quantitative PCR. Each value is reported as the mean ± standard deviation and is representative of results obtained from three independent experiments. (B) MDA-MB-231 and HS578T cells were incubated with cycloheximide (10 µg/ml), and/or ACF (10 µM) and after 0, 1 and 2 h the expression of MCL-1 was determined using western blot analysis. (C) MDA-MB-231 and HS578T cells were incubated with ACF and proteasome inhibitor, MG-132, and expression of MCL-1 was determined using western blot analysis. (D) HS578T cells were treated with 10 µM ACF and and/or MG-132, and after 6 h MCL-1 was pulled down and the ubiquitination level was analyzed using western blotting. ***P<0.001 compared with non-treated cells. ACF, acriflavine; MCL-1, myeloid cell leukemia sequence 1.