741985.fig.003a
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741985.fig.003b
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741985.fig.003c
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Figure 3: REM inhibits YB-1-dependent MDR1 expression. (a) YB-1 localization. Cells were stained with anti-YB-1 antibody (green), phalloidin (red), and TOPRO-3 (blue) to visualize YB-1 intracellular localization, actin, and nucleus. Scale bars, 50 μm. The objective, 20X. (b) MCF-7/Dox cells were treated with the indicatives for 6 hours and then fixed with 1% formaldehyde. Fragmented DNAs were incubated with the appropriate antibodies, and then PCR was routinely performed. GAPDH was used as an internal control. Relative values of YB-1 enrichment normalized by ddCT were obtained by experiments performed in triplicate. -value less than 0.05 (marked with an asterisk, *) was considered statistically significant. (c) MCF-7/Dox cells were cotransfected with MDR1-luc and YB-1 constructs for 24 hours and then treated with the indicatives for another 6 hours. Assays were performed in triplicate, and value less than 0.05 (marked with an asterisk, *) was considered statistically significant.