Fig. 5. TGF-ß1-induced EMT in NMuMG cell line reciprocally modulates paxillin and Hic-5 protein expression. A, Detergent soluble lysates from cells either unstimulated (–) or stimulated (+) with 2 ng/ml TGF-ß1 for 48 hours were processed for western blot analysis. Paxillin monoclonal antibody PXC10 was used to recognize both and forms of paxillin. Paxillin expression is suppressed while Hic-5 expression increases following a TGF-ß1-induced EMT in the NMuMG cell line. E-cadherin and N-cadherin expression were monitored to verify the induction of an EMT. (B) Western blot analysis of detergent-soluble lysates obtained from NMuMG cells either unstimulated (–) or stimulated (+) with 2 ng/ml TGF-ß1 for 48 hours. Phospho-specific antibodies to paxillin Y31 and Y118 residues as well as to FAK Tyr397 were utilized. Paxillin Y31 and Y118 phosphorylation occurs concurrent with the suppression of paxillin following an EMT.