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. 2016 Sep 1;7(40):66087–66099. doi: 10.18632/oncotarget.11807

Figure 5. Down-regulation of RORα attenuates MLN4924-induced cell growth suppression.

Figure 5

A. U2OS osteosarcoma cells were transfected with two distinct RORα-specific siRNAs or a negative control siRNA. 72 h after transfection, whole cell lysates were analyzed with Western blot using an antibody against RORα. B. U2OS cells were treated with MLN4924 (3 μM) or DMSO for 48 h after transfection with RORα-specific siRNAs or negative control siRNA, and the cell proliferation was monitored with an MTT assay at 48 and 72 h after MLN4924 treatment. Each value is the mean ± SEM of three replicates from a single assay (**P < 0.01 by post hoc Bonferroni t test). C. U2OS cells transfected with RORα-specific siRNA or negative control siRNA after MLN4924 (1 μM) treatment for 0, 24 and 48 h. Cells were harvested and fixed in ice-cold 70% ethanol overnight at −20°C, then stained with PI (5 μg/100 μL) for 30 min at 4 °C in the dark. DNA profiles were analyzed by flow cytometry. D. The graph illustrates the percentage of > 4N cell of four replicates from a single assay (**P < 0.01 by post hoc Bonferroni t test).