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. 2016 Feb 24;7(13):16619–16635. doi: 10.18632/oncotarget.7672

Figure 6. FOXM1 regulates EF2K mRNA and protein expression.

Figure 6

Cells were transfected with 50 nM FOXM1#1 or FOXM1#2 siRNA or control siRNA. Protein extracts were isolated 72 h after transfection. eEF2K protein levels were determined by Western blot analysis using anti-eEF2K monoclonal antibody. β1-Actin was used as a loading control. siRNA-mediated silencing of FOXM1 inhibited eEF2K expression in both MDA-MB-231 (A, B) and BT-20 (C, D) cells. Total RNA was isolated 72 h after transfection. eEF2K mRNA levels were determined by standard RT-PCR. Band intensities indicated obvious down-regulation of eEF2K transcription in both MDA-MB-231 (E, F) and BT-20 (G, H) cells. GAPDH was used as a loading control. In addition, MDA-MB-231 cells were transfected with a FOXM1 expression vector (400 ng) or an empty vector. eEF2K (I, J) and FOXM1 (K, L) protein expression levels were determined by Western blot analysis. β1-Actin was used as a loading control. The data are means with standard deviations. *represents significant difference between indicated groups.