Anti rhoa antibody

Human integrin β₁ cDNA was cloned into plenti6/V5-DEST vector after digestion with EcoRI and XhoI. Integrin E-to-A mutants were generated using PCR and cloned into plenti6/V5-DEST vector by EcoRI and XhoI. GST-β₁CD and recombinant Gα₁₃ purification was described previously (Shen et al., 2013 (link)). Anti-RhoA antibody was from Cytoskeleton (Denver, CO); anti-Gα₁₃(sc410), anti–c-Src (sc18), anti–integrin β₁ K20 (sc18887), and anti–integrin β₁ JB1B (sc59829) antibodies were from Santa Cruz Biotechnology (Dallas, TX); anti-Gα₁₃ (26004) was from NewEast Biosciences (King of Prussia, PA); anti–phospho-Src Y⁴¹⁶ antibody was from Cell Signaling Technology (Danvers, MA); anti–GST tag antibody and Alexa Fluor 555 conjugate were from EMD Millipore (Billerica, MA); Lipofectamine 2000, ViraPower Lentiviral Expression System, and Alexa Fluor 546–conjugated phalloidin were from Invitrogen (Carlsbad, CA); fibronectin was from BD Biosciences (Franklin Lakes, NJ); the Active Rho Pull-Down and Detection Kit was from Pierce, Thermo Scientific (Waltham, MA).

Myristoylated G13BP and scrambled peptide were synthesized and purified at the Research Resource Center at University of Illinois, Chicago. Anti-Gα13 (sc-410), anti-Src (sc-18, sc-5266), and anti–VE-cadherin (sc-9989 and sc-6458) antibodies, as well as human VE-cadherin siRNA were purchased from Santa Cruz Biotechnology; mouse monoclonal anti-Gα13antibody (26004) was from Neweast Biosciences; anti–phospho-Src Y416 antibody was obtained from Cell Signaling Technology; anti–phospho-VE-cadherin Y658 antibody, Src kinase assay kit, and anti–v-Src antibody were obtained from Millipore; anti–β-actin and anti-Flag (M2) antibodies, TNF, GDT and GTPγS, NAC, and DPI were purchased from Sigma-Aldrich; Lipofectamine 2000, anti-GFP antibody, and Lipofectamine 2000 were purchased from Invitrogen; HMVEC-L and Amaxa Nucleofector kit (VPB-1002) were obtained from Lonza; and HisPur Ni-NTA Spin Purification kit was obtained from Pierce. SensoLyte ADHP Hydrogen Peroxide Assay kit was from purchased AnaSpec. Anti-RhoA antibody and GST-Rhotekin beads were purchased from Cytoskeleton Inc.