Figure 2. Depletion of km23-1 in RKO human CRC cells inhibits transactivation of the proximal AP-1 site in the TGFβ1 promoter, TGFβ1 secretion, c- Fos-DNA binding to the TGFß1 promoter, and Elk-1 activation.

A: RKO stable transfectants were transiently transfected with phTG5-Lux and luciferase assays were performed as described previously [20], [40]. Data are representative of three independent experiments. B: TGFβ1 concentrations in CM from RKO stable transfectants were measured by ELISAs as described previously [20], [40]. *p<0.01 compared to the NC siRNA. C: ChIP assays were performed [20], [40] using either the human TGFβ1 promoter region from −410 to −260 (upper panel) or the hTERT promoter region from −1734 to −1584 (lower panel) with the indicated antibodies. Input: Equal samples of chromatin DNA without prior immunoprecipitation. D: Western blotting of phospho- and total protein expression levels for Elk-1 in RKO human CRC cells. Three independent experiments were performed and representative figures are shown.