NDR1 represses TGFβ-induction of endogenous PAI-1 gene expression.

<p><b>A.</b> RNA extracts of untreated or TGFβ-treated NMuMG cells transfected with a control RNAi plasmid or the combination of NDR1i-1 and NDR1i-2 RNAi plasmids were subjected to quantitative RT-PCR to determine the abundance of PAI-1 mRNA, where GAPDH mRNA was used as an internal control. Data are presented as the mean+SEM (n = 5) of GAPDH-normalized PAI-1 mRNA abundance relative to untreated control. <b>B.</b> Lysates of NMuMG cells expressing FLAG-tagged wild type NDR1 (WT) or kinase-inactive NDR1 in which Lysine 118 is mutated to arginine (KR), or vector control were subjected to immunoblotting using the NDR1 or actin antibody, with the latter serving as a loading control. <b>C.</b> RNA extracts from untreated or TGFβ-treated NMuMG cells expressing wild type or kinase-inactive NDR1 or the vector control were subjected to quantitative RT-PCR analysis of PAI-1 and GAPDH mRNA as described in A. Data are presented as the mean+SEM (n = 3) of relative GAPDH-normalized PAI-1 mRNA abundance as in A. *, **, or *** indicates significant difference from the TGFβ-treated control at p<0.05, p<0.01, or p<0.001, respectively (ANOVA).