Desmoplakin

Western blotting was performed according to the standard protocol. The primary antibodies, including GTSE1(Proteintech, 21,319–1-AP), Ki67(Proteintech, 27,309–1-AP),mutant p53(Abcam, ab32049), p53(Proteintech, 10,442–1-AP),Cyclin D1(Proteintech, 60,186–1-Ig),Cyclin E1(Proteintech, 11,554–1-AP), p21(Proteintech,60,214–1-Ig), E-Cadherin (Proteintech, 20,874–1-AP), Desmoplakin (Proteintech,25,318–1-AP), N-Cadherin (Proteintech,22,018–1-AP), Vimentin (Proteintech, 10,366–1-AP), Nanog (Proteintech, 14,295–1-AP), ABCG2(CST, 42078S), TAZ (CST, 4883S), YAP (CST, 14074S), ERK1/2(Affinity, AF0155), phospho-ERK1/2(Affinity, AF1015), AKT (CST, 2938S), Phospho-Akt (Thr308) (CST, 13038S), Phospho-Akt (Ser473) (CST, X4060S), FoxC2(CST, 12974S),Slug (CST, 9585 T),Twist1(CST, 46702S),Snail (Proteintech, MG-3879 T) and GAPDH (Proteintech, 60,004–1-Ig) were used at a dilution of 1:1000.

Cells were lysed with RIPA lysis buffer (Beyotime) and added the protease inhibitor. Next, the concentration of proteins was measured by a BCA Protein Quantification Kit (Yeasen). The standard western blotting protocol was followed. The primary antibodies included antibodies specific for EMT markers, Desmoplakin (Proteintech, # 25318-1-AP), ZEB2 (Proteintech, # 14026-1-AP), N-cadherin (Proteintech, # 22018-1-AP), E-cadherin (Proteintech, # 20874-1-AP), Vimentin (Proteintech, # 10366-1-AP), Slug (Signalway Antibody, #24463), Snail (Cell Signaling Technology, # 3879 T), GAPDH (Proteintech, #10494-1-AP), N-WASP (Cell Signaling Technology, # 4848 T), Phospho-N-WASP (Affinity Biosciences, # AF7032), Cortactin (Proteintech, # 11381-1-AP), SF3B3 (Abcam, # ab209402), IKKα (Cell Signaling Technology, #11930), IKKβ (Cell Signaling Technology, #8943), Phospho-IKKα/β (Cell Signaling Technology, #2697), Phospho-p65 (Cell Signaling Technology, #3033), IκBα (Cell Signaling Technology, #4814), Phospho-IκBα (Cell Signaling Technology, #2859), and p65 (Cell Signaling Technology, #8242) were detected. The antibody dilution ratios were determined according to the protocol. The reference gene was GAPDH.

Primary antibodies against the following antigens were used: K1, K14, K15, K19, E-cadherin, Cdc42, AP-1, p-PAK1 (Abcam, UK), K6, loricrin, involucrin (Covance, USA), β-catenin, bromodeoxyuridine (BrdU; Santa Cruz, USA), α6-integrin (BD Biotech, USA), F4/80 (eBioscience, USA), SPRR1A (Biorbyt, UK), SPRR1B (Aviva Systems Biology, USA), desmoplakin, ZO-1, SPRR2D, PAK1, cyclin D1, caspase 3, occludin and SPRR2G (Proteintech, China), Bim, p-Bim, Bcl-2, p-Bcl-2, JNK, p-JNK, p38, p-p38, ERK, and p-ERK (Cell Signaling, USA).