Anti histone 2b

Cells were lysed in 1× SDS lysis buffer (150 mmol/L NaCl, 25 mmol/L Tris-HCl, pH 7.6, 1% sodium deoxycholate, and 1% NP-40) in the presence of a protease inhibitor cocktail (Roche). Nearly 20 μg of cell lysate was isolated with SDS-PAGE and then transferred onto a PVDF membrane (Millipore, Billerica, MA, USA). The PVDF menbranes were then blocked with 10% skimmed milk for 30 min. Immunoblot analysis was performed with the following primary antibodies: anti-α-Tubulin, anti-COX-2, anti-Histone 2B, anti-IκBα, anti-iNOS and anti-p-p65 antibodies (Abcam, Cambridge, UK); anti-IKK and anti-p-IKK antibodies (Cell Signaling Technology, Danvers, MA, USA); anti-p65 antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-GAPDH antibodies (Millipore, Billerica, MA, USA). Dilution and catalog number of each antibody were listed in Supplementary Table S1 in Supplementary Material. The secondary antibodies, sheep anti-rabbit or anti-mouse IgG-HRP, were from Thermo Fisher Scientific (Waltham, MA, USA). The proteins were visualized using an ECL detection kit (Thermo Fisher Scientific).

Anti‐GSK3β (#22104‐1‐AP), anti‐Wnt2 (#27214‐1‐AP), and anti‐β‐catenin (#51067‐2‐AP) antibodies were purchased from Proteintech (Chicago, IL). Anti‐pSer1490‐LRP6 (#2568) and anti‐LRP6 (#3395) were from Cell Signaling Technology (Beverly, MA). Anti‐β‐tubulin (#ZS‐9104) was purchased from Zhongshan Technology (Beijing, China). Anti‐Histone 2B (#GR193918‐1) was purchased from Abcam (Shanghai, China). Anti‐p37 monoclonal antibody PD4 was generated and characterized previously.2, 5 FITC‐conjugated and HRP‐conjugated secondary antibodies were from Zhongshan Technology (Beijing, China). XAV939 was purchased from Selleck (Houston, TX).