Mouse anti ph2a ser139

The protocols of the western blot were performed as described by Hanson et al. [26 (link)].
Protein extracts, quantified by a Bradford Protein Assay (Bio-Rad Laboratories, CA, USA), underwent SDS-polyacrylamide gel electrophoresis and were transferred to Immobilon-P membranes. The following antibodies were used: rabbit anti-NF-κB, rabbit anti-βcatenin, goat anti-matrin3, goat anti-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1 : 500; rabbit anti-cyclin E2, cyclin D1, cyclin B1, p21, Pmyt1, Oct4, and mouse anti-cyclin A1, and SSEA-4 (Cell Signalling Technology, Beverly, MA, USA), mouse anti-tubulin, and mouse anti-sc-35 (Sigma-Aldrich St. Louis, MO, USA), rabbit anti-Nrf2 (Abcam, Cambridge, UK), rabbit anti-Nox4 (Novus Biologicals, CO, USA), and mouse anti-pH2A (Ser139), mouse anti-CD90 and anti-CD105 (Millipore, Billerica, MA, USA) rabbit anti-CD73 (Genetex, Irvine, CA, USA), diluted 1 : 1000; peroxidase-labelled anti-rabbit, mouse, and goat secondary antibodies diluted 1 : 3000 (Pierce Antibodies, Thermo Scientific; Rockford, IL, USA). Ab dilution was performed in TBS-T pH 7.6 containing 3% BSA. The membranes were visualized using Supersignal substrate chemiluminescence detection kit (Pierce, Rockford, IL, USA). Anti-actin antibody was used as control of protein loading.

The protocols of the Western blot were performed as described by Hanson et al. [32 (link)]. Briefly, protein extracts, quantified by a Bradford Protein Assay (Bio-Rad Laboratories, CA, USA), underwent SDS-polyacrylamide gel electrophoresis and were transferred to Immobilon-P membranes. The following antibodies were used: rabbit anti-ERK1/2, goat anti-Matrin3, goat anti-βactin, anti-p22phox (Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1 : 500, rabbit anti-Akt1, rabbit anti-Rac1, and rabbit anti-ERK1/2 (Cell Signalling Technology, Beverly, MA, USA), mouse anti-tubulin, rabbit anti-Nox1, and mouse anti-sc-35 (Sigma Aldrich St. Louis, MO, USA), rabbit anti-Nox4 (Novus Biologicals, CO, USA), rabbit anti-Nox2, and mouse anti-pH2A(Ser139) (Millipore, Billerica, MA, USA) diluted 1 : 1000; peroxidase-labelled anti-rabbit, mouse and goat secondary antibodies diluted 1 : 3000 (Pierce Antibodies, Thermo Scientific; Rockford, IL, USA). Ab dilution was performed in TBS-T pH 7.6 containing 3% BSA. The membranes were visualized using Supersignal substrate chemiluminescence detection kit (Pierce, Rockford, IL, USA). Anti-βactin antibody was used as control of protein loading. Quantization of the signal was obtained by chemiluminescence detection on a Kodak Image Station 440CF and analysis with the Kodak 1D Image software.