Plakoglobin

Skin equivalents were fixed in 10% buffered formalin and embedded in paraffin. Sections were stained with H&E and histopathological evaluation of skin equivalents was performed by two specialists in pathological anatomy: MJFA, specialized in human pathology and AB, a veterinarian expert in animal pathology. Immunostaining was performed using antibodies against IKKα NB-100-56704 (Novus Biologicals, Cambridge UK); IKKα sc-7182 (Santa Cruz Biotechnology, Inc. Heidelberg, Germany); IKKα 556532, Plakoglobin (BD Bioscience, NJ, USA); Involucrin, Filaggrin (Covance, CA, USA); BrdU (Roche, Mannheim, Germany). Sections were incubated with a biotinylated secondary antibody, and then with streptavidin conjugated to horseradish peroxidase (DAKO A/S, Glostrp, Denmark). Antibody localization was determined using 3,3-diaminobenzidine (DAB) in H₂O (Vector Laboratories; Burlingame, CA, USA).
Pressure cooker with DAKO target retrieval solution ph9.0 (DAKO) was employed for antibodies detection.

A polyclonal antibody to SNX27 was generated as described previously [11 (link)]. A monoclonal antibody against fusion proteins tagged with c-Myc (clone 9E10) and polyclonal giantin antibody (# ab93281) were purchased from Abcam (Cambridge, MA). β-Catenin, E-cadherin, GSK3, EEA1 and plakoglobin monoclonal antibodies were purchased from BD Biosciences (Franklin Lakes, New Jersey). The Cell Light™ early endosome (RFP-Rab5a) marker was purchased from Invitrogen and used at 25m.o.i. A mouse monoclonal antibody to Lef-1 (# 17-604) was purchased from Millipore (Billerica, MA).