Anti p120 catenin

A six-well plate was used to culture HaCaT cells or NHEK cells for 24 h, followed by 12 h, 24 h, 48 h, or 72 h of UPF treatment at the above-mentioned concentrations along with controls. Following cell harvest, the total protein extracted from each culture was resolved on 8% SDS-PAGE. This was followed by transfer to polyvinylidene difluoride (PVDF) membrane and overnight incubation with the following antibodies at 4 °C: with rabbit anti-phospho-p120-catenin, anti-p120-catenin, anti-phospho-PLCγ1, anti-PLCγ1, anti-phospho-ERK1/2, anti-ERK1/2, anti-phospho-p38, anti-p38, anti-CaSR, anti-Involucrin or anti-tubulin antibody (Cell Signaling Technology, Beverly, MA, USA), and with anti-Filaggrin (Santa Cruz Biotechnology, Dallas, TX, USA) for overnight. Post addition of goat anti-rabbit antibody or goat anti-mouse antibody (Cell Signaling Technology, Beverly, MA, USA) respectively, the detection was done using a chemiluminescence substrate (Pierce, Rockford, IL, USA) followed by the use of Amersham Imager (GE Healthcare Biosciences, Pittsburgh, PA, USA) to record images.