Mouse Actn4 cDNA was subcloned into the pmApple and pEGFP vectors. Expression construct of Shp2 were subcloned into pflag-CMV2 vector. Expression construct of mcherry-ACTN1 was obtained from JC Kuo (NYCU). Expression construct of Shp2-SWAP FRET biosensor was provided by Dr. Yingxiao Wang (UCSD) (Sun et al, 2013 (link)). Anti-paxillin antibody and mouse collagen type-IV from BD Biosciences; anti-β-actin, anti-β-tubulin, and anti-flag antibodies, FITC-phalloidin, ROCK inhibitor Y27632, and Shp2 inhibitor IIB-08 from Sigma-Aldrich; FAK inhibitor 14 from Tocris Bioscience, anti-pY397FAK, anti-pY31-paxillin, and anti-ZO-1 antibodies from Invitrogen; anti-pY542 Shp2, anti-pT18/pS19-MLC, and anti-MLC antibodies from Cell Signaling; anti-α-actinin-1 antibody from Chemicon; anti-α-actinin-4 and anti-phosphotyrosine (clone 4G10) antibodies from Millipore; anti-FAK, anti-Shp2, anti-ROCK1, and anti-ROCK2, antibodies from Santa Cruz Biotechnology Inc.; anti-synaptopodin antibody from Novus; anti-podocin antibody from Abcam; λ protein phosphatase from New England BioLabs. Anti-pS1333 ROCK1 and anti-pS1366 ROCK2 antibodies were generated in our laboratory (Chuang et al, 2012 (link), 2013 (link)).
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