TWIST1 induces IL8 Production. (A) Cytokine array blotted with conditioned media collected from MCF10A or MCF10ATw cell cultures. Cytokines detected in MCF10ATw conditioned medium are shaded in Additional File 2, Table S1. POS, positive controls. (B) Relative mRNA levels of TWIST1 and IL8 normalized to β-ACTIN See Methods) in MCF10A cells 48 h post-transduction with adenoviruses that express TWIST1 (Ad.TWIST) at a serial multiplicity of infection (MOI). Cells transduced with Ad.EGFP (enhanced green fluorescent protein) at an MOI of 10 were used as controls. (C) Relative mRNA levels of TWIST1 and IL8 normalized to β-ACTIN as in (B) in SKBR3 and MCF7 cells 48 h post-transduction with Ad.TWIST as compared to Ad.EGFP (control). MOI = 5. (D) Relative mRNA levels of IL8, normalized as in (B), in BT549 cells with TWIST1 KD (right). Cells were transduced with lentiviruses that expressed a non-targeting shRNA (shCtrl) or shRNAs that targeted TWIST1 (shTw1, shTw2). Western blot (left) shows the levels of TWIST1 under these conditions. One and four minute exposures of the blot are shown. RELA is shown as a KD control. The data are from single representative quantification or images of at least three independent experiments.