Anti phospho akt s473 and anti akt pan

An anti-human APC-CD271 antibody (clone ME20.4-1.H4, Miltenyi Biotec, Germany) was used for flow cytometry analysis. The following antibodies were used for Western blotting analysis: anti-phospho-p44/42 MAPK (ERK1/2, Thr202/Tyr204) (20G11, Cell Signaling Technologies, Danvers, MA, USA), anti-ERK (137E5, Cell Signaling), anti-phospho-NF-κB p65 (S536) and anti-NF-κB p65 (93H1 and D14E12, Cell Signaling), anti-phospho-Iκbα (Ser32) and anti-Iκbα (14D4 and L35A5, Cell Signaling), anti-phospho- Akt (S473) and anti-Akt (pan, Cell Signaling) anti-α-tubulin (B-5-1-2, Santa Cruz Biotechnology, Dallas, TX, USA), anti-involucrin (clone SY-5, Sigma-Aldrich, St. Louis, IL, USA), anti-CDKN1C (NBP1-89917, Novus Biologicals, Littleton, CO, USA), and anti-β-actin (AC-15, Sigma-Aldrich). The following antibodies were used for immunohistochemistry analysis: anti-CD271 (C40-1457, BD Biosciences, Franklin Lakes, NJ, USA), anti-Ki67 [clone 30-9 (Roche, Switzerland) for DAB staining and clone SP6 (Abcam, UK) for immunofluorescence staining] and anti-involucrin (clone SY-5, Sigma-Aldrich).