Non phospho active β catenin ser33 37 thr41

The following antibodies were used in Western blotting and immunofluorescence:
phospho-GSK3α/β (CST 8566), phospho-β-Catenin (Ser33/37/Thr41) (CST9561), non-phospho (Active) β-Catenin (Ser33/37/Thr41) (CST 8814), total β-Catenin (CST 9587), phospho-cyclin D1 (CST 3300), PPARγ (CST 2443), p27 (CST 2552), Ki-67 (CST 9129), phospho-c-Jun (CST 9164), c-Myc (CST 13987), Cytochrome c (CST 4280),GFAP (CST 3670), vimentin (CST 5741), NF-kB (CST 8242), c-Fos (CST 2250) from Cell Signaling Technology (Danvers, MA, US); Lamin B1 (10H34L18), Cyclin D1 (MA5-14512), Bcl-2 (13-8800), phospho-CREB (MA1-083), Ki-67 (MA5-14520) from Invitrogen, Thermo Fisher Scientific, and Phospho-JNK (sc-6254) from Santa Cruz Biotechnology (Dallas, TX, USA).

Western blot assays were conducted as previously described (Xu et al., 2014 (link)). The proteins were extracted using a radioimmunoprecipitation assay buffer (Sangon Biotech), separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to polyvinylidene difluoride membranes. The membranes were then blocked for 1 h and incubated with the indicated antibody in PBS plus 5% bovine serum albumin overnight at 4 °C. After washing with PBS plus 0.1% Tween-20, the membranes were incubated with horseradish peroxidase-conjugated goat anti-mouse or anti-rabbit immunoglobulin G (Sangon Biotech) for 1 h. After subsequently washing, the immunoreactive bands were visualized using the Tannon 2500 imaging system, and the densitometry of the bands was analyzed using ImageJ software. Primary anti-human antibody against USP5 was purchased from Abcam (Cambridge, MA, USA), and E-cadherin, N-cadherin, vimentin, β-catenin, non-phospho (Active) β-catenin (Ser33/37/Thr41), glycogen synthase kinase-3β (GSK-3β), phospho-GSK-3β (Ser9), ubiquitin, and GAPDH were all purchased from Cell Signaling Technology (CST, Danvers, MA, USA).