Anti phospho gsk3β ser9

The following antibodies were used: Anti-YAP (sc-101199), anti TEF-1 (sc-376113), Anti-GSK3β (sc-9166), Anti phospho GSK3β Ser9 (sc-11757), Anti β-catenin (sc-7199) (Santa Cruz Biotechnology). Anti-Lats1 (3477), Anti-phospho Lats1 Ser909 (9157), Anti-Smad2/3 (8685) and Anti-phospho-YAP (S127) (4911), Anti β-actin (3700) (Cell Signaling). Alexa Fluor 488-conjugated secondary antibodies (Invitrogen). The chemicals were used in this study: GF 109203X (B6292), Go6976 (G1171), phorbol 12-myristate 13-acetate (PMA) (Sigma, P1585) were purchased from sigma. Lapatinib (S1028), Erlotinib (S1023) were purchased from Selleckchem. Recombinant human transforming growth factor 1 (TGF-β) (PHG9204) was purchased from Life technologies. The plasmids were used: 8xGTIIC-luciferase (#34615), YAP-GFP (12), pEGFP-N1 (Clontech), pGL4 (Promega), human B-catenin pcDNA3 (#16828), c-Flag pcDNA3 (#20011), TOP flash and FOP flash were gifted from Dr. Arthit [50 (link)].

We evaluated the regulatory effects of cyclin G2 on the canonical Wnt signalling pathway and on the expression of proteins related to renal injury. Protein concentrations were detected with BCA (Life Technologies). Primary antibodies were anti–collagen IV, anti–cyclin D1, anti–β-tubulin, anti–phospho-β-catenin (Ser33/37/Thr41) (Cell Signalling Technology, Boston, MA), anti-GSK3β, anti–phospho-GSK3β (Ser9) (Santa Cruz Biotechnology), anti–β-catenin (Sigma-Aldrich), anti-fibronectin (FN), anti-MMP7, and anti–cyclin G2 (Proteintech, Chicago, IL). All experiments were performed in triplicate.