Figure 6.

EMT-inducing transcription factors regulate ABC transporter expression. (a) Graph represents luciferase assay undertaken in MCF7 cells to assess the ABCC5 promoter activity (ABCC5-Luc; firefly luciferase) on co-transfection with pRL-TK (Renilla luciferase) and pBp-empty vector/Snail/mTwist/FOXC2. After 48 h culture, firefly luciferase activity was measured and normalized to Renilla luciferase activity and depicted as relative luciferase units (RLU), n=3. Error bars denote ±S.E.M. ^*P<0.05; ^**P<0.01. (b) Schematic of E-box elements in 2.2 kb long promoter regions of ABCC4 and ABCC5 genes. (c) Gel picture represents PCR carried out on ChIP undertaken in MCF7 cells transiently overexpressing Flag-Twist using anti-Flag Ab. PCR was carried out with the eluted DNA fragments using primers depicted in (b). No antibody (No Ab) served as a negative control and rabbit IgG (Rab IgG) served as an isotype control. Control primers were designed ∼700 bp upstream of E-box elements in the ABCC5 promoter and served as a specificity control