Figure 2

TGFβ mediates its effects through LIF regulation. A, WM278 cells were treated or not with TGFβfor 24 h and LIF expression was analyzed by qPCR. Error bars are standard deviations and z-test was performed (***p < 0.001). B, WM278 and WM793B cells were treated or not with TGFβ for 24 h and LIF expression was analyzed by Western blot (left panel) after concentration of conditioned media. β-tubulin was used as control. Right panel: Densitometric analysis of LIF protein levels. Error bars are standard errors of mean and t-test was performed compared to non-treated control (***p < 0.001). C, WM278 cells transfected with scrambled or LIF siRNA 48 h earlier were treated or not with TGFβ for 24 h and apoptosis was determined by caspase3/7 activity. Error bars are standard errors of mean and z-test was performed compared to non-treated control (**p < 0.01, ***p<0.001). D, WM278 cells transfected with scrambled or LIF siRNA 48 h earlier were treated or not with TGFβ for 24 h and apoptosis was determined by caspase3/7 activity. Data is graphed as the geometric mean of relative luciferase units normalized to non-treated control for at least 3 independent experiments. Error bars are the standard errors of mean and z-test was performed compared to non-treated control (**p < 0.01). E, WM278 cells transfected with scrambled or LIF siRNA 48 h earlier were treated or not with TGFβ for 24 h. LIF expression was analyzed by qPCR. Data is graphed as the mean of fold induction of gene expression in response to TGFβ for at least 3 independent experiments. Error bars are the standard errors of mean and t-test was performed compared to mock and scrambled siRNA treated conditions (**p < 0.01).