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.

<p><b>A:</b> RKO stable transfectants were transiently transfected with phTG5-Lux and luciferase assays were performed as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Liu1" target="_blank">[20]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Pandey1" target="_blank">[40]</a>. Data are representative of three independent experiments. <b>B:</b> TGFβ1 concentrations in CM from RKO stable transfectants were measured by ELISAs as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Liu1" target="_blank">[20]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Pandey1" target="_blank">[40]</a>. *p<0.01 compared to the NC siRNA. <b>C:</b> ChIP assays were performed <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Liu1" target="_blank">[20]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066439#pone.0066439-Pandey1" target="_blank">[40]</a> 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. <b>D:</b> 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.