Rabbit anti phospho p120 catenin

The antibodies used for immunoblot analysis in this study are as follows. All antibodies were used at a dilution of 1:1000 in TBS-T (20 mM Tris-HCl, pH 7.6, 137 mM NaCl, 0.2% [v/v] Tween-20) with 3% (w/v) BSA unless otherwise indicated. Rabbit anti-pTyr (Cat#8954), mouse anti-HisTag (Cat#2366), rabbit anti-Paxillin (Cat#12065), rabbit anti-phospho-Paxillin (Y118; Cat#2541), rabbit anti-phospho-p120-Catenin (Y228; Cat#2911), and rabbit anti-phospho-p120-Catenin (Y904; Cat#2910) primary antibodies were all purchased from Cell Signalling Technology. Mouse anti-Afadin (Cat#610732) and mouse anti-p120-Catenin (Cat#610133) primary antibodies were purchased from BD Transduction Laboratories. Mouse anti-PTPRM (PTPRU cross-reactive; Cat#sc-56959)^{21 (link)} primary antibody was purchased from Santa Cruz. Rabbit anti-PTPRK primary antibody was generated in a previous study^{3 (link)}. Mouse anti-alpha-tubulin (Cat#T6199) and anti-FLAG (Cat#F7425) primary antibodies were purchased from Sigma Aldrich. HRP-conjugated anti-mouse (Cat#711-035-151) and anti-rabbit (Cat#711-035-152) secondary antibodies (1:5000 in TBS-T) were purchased from Jackson Immuno-Research.

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.