Mouseanti vimentin monoclonal antibody clone v9

Pyloric atresia junctional epidermolysis bullosa (PA-JEB) keratinocyte cells expressing β4 integrin (PA-JEB/β4) were cultured as described previously (Geerts et al., 1999) . For immunofluorescent analysis, keratinocytes grown on cover slips were fixed with 10% MeS buffer (100 mM MeS, pH 6.9, 1 mM EGTA and 1 mM MgCl2) and 90% methanol for 5 min on ice. After blocking with 5% bovine serum albumin for 1 hour, the cells were incubated with mouse anti-vimentin monoclonal antibody (Clone V9-Dako) and rabbit anti-keratin 14 polyclonal antibody (Covance) for 1 hour. Subsequently all the cells were incubated with goat anti-rabbit and goat antimouse antibodies (Alexa 488 and Alexa 555 Invitrogen) for 30 min. All the fixation and staining steps were done at room temperature. Primary human umbilical vein endothelial cells (HUVECs) were cultured, fixed and stained as described before (Nieuwenhuizen et al., 2015; Nahidiazar et al., 2015) .