Figure 5

YB-1 knockdown induces different gene expression patterns in RU and RR cells. (A) Quantitative-RT-PCR analyses of relative mRNA transcripts of Sox2 only target NANOG in MCF7 and ZR751 RU and RR cells after scrambled or YB-1 siRNA treatment at 20 nM for 72 hours. (B) MCF7 RU and RR cells were transfected with 3 μg of pcDNA3-Flag-Empty Vector (EV) or pcDNA3-Flag-Sox2 and harvested for mRNA after 72 hours. Q-PCR analyses was performed using primers designed against SOX2 and NANOG. SOX2 transcript levels are normalized to the Flag-EV transfections. Accompanying Flag western blot is shown. (C) Quantitative-RT-PCR analyses of Sox2 and YB-1 targets CCND1 (Cyclin D1), and ITGA6 (CD49f) in MCF7 and ZR751 RU and RR cells after scrambled or YB-1 siRNA treatment at 20 nM for 72 hours and (D) accompanying western blot showing YB-1, Sox2 and Cyclin D1 protein expression.