Figure 4
From: The cancer gene WWOX behaves as an inhibitor of SMAD3 transcriptional activity via direct binding
WWOX inhibits TGFβ-dependent transcription and decreases SMAD3 occupancy at target gene promoters. (A) 3TP-LUX luciferase reporter assay in MCF10 cells treated with or without doxycycline to induce WWOX expression and with or without TGFβ1 for 8 hours as indicated. Experiment done in triplicate. (B-C) Chromatin immunoprecipitation of SMAD3 (or IgG control) followed by qPCR in MCF10 cells transiently infected with the doxycycline-inducible WWOX expression lentivirus system. Primers used for qPCR span SMAD binding elements in the ANGPTL4 promoter (B) or the SERPINE1 promoter (C). Error bars represent SD for all graphs.