Anti glyceraldehyde 3 phosphate dehydrogenase gapdh

Rabbit anti-PA, anti-PB1, anti-PB2, and anti-NP antibodies were purchased from GeneTex Inc. (GeneTex, United States). Mouse monoclonal antibodies against influenza A virus HA and M2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was purchased from Beyotime Biotechnology. The antibodies of human RNA pol II, RNA pol II CTD phosphor Ser2 and RNA pol II CTD phosphor Ser5 were purchased from Active Motif company.

To investigate the effects of BMP9 on the protein expression of ALP, OPN, OCN, p-Smad2/3 and p-JNK, we incubated the BMP9-infected hPDLSCs in the presence or absence of different concentrations of SP600125 or co-infected with AdR-si-JNK. Cells were lysed in a radioimmunoprecipitation assay (RIPA) buffer (Beyotime, Shanghai, China). After centrifugation, proteins in the supernatant were collected and quantified using a bicinchoninic acid (BCA) protein assay kit (Beyotime, Shanghai, China). The total proteins were denatured by boiling for 5 min, resolved by 10% gradient SDS-PAGE and then transferred to PVDF membranes. After blocking with 5% BSA for 2 h, the membranes were incubated with 1:2000 primary antibodies against anti-ALP, anti-OPN, anti-OCN, anti-p-Smad2/3, anti-p-JNK1/2 (all from Abcam, Cambridge, UK) and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as housekeeping gene (Beyotime, Shanghai, China) overnight, followed by the addition of HRP AffiniPure goat anti-rabbit IgG(H+L) (Abcam, Cambridge, UK) as the secondary antibody for 2 h. The blots were visualized by enhanced chemiluminescence (ECL) and autoradiography.