Rabbit polyclonal anti cyr61

Equal amounts of whole joint tissue were lysed in lysis buffer, and tissue proteins were extracted. The cells were lysed, and the supernatant was used for protein quantification. GAPDH was used as a protein loading control. The following primary antibodies were used in the subsequent experiment: rabbit anti-IL-1β, rabbit anti-TNF-α, mouse anti-IL-6 (all from Abcam, Cambridge, MA, USA), rabbit polyclonal anti-Cyr61 (Invitrogen, Carlsbad, CA, USA), and rabbit anti-GAPDH (Cell Signaling Technologies, Beverly, MA, USA). The secondary antibodies used were goat anti-rabbit IgG HRP, goat anti-mouse IgG HRP, and donkey anti-goat IgG HRP (Beyotime Biotechnology, Shanghai, China). The target proteins were examined using a Tanon-5200 imaging system (Tanon Science and Technology Corporation, Shanghai, China) and visualized with autoradiography film.

Equal amounts of whole joint tissue were lysed in lysis buffer, and tissue proteins were extracted. The cells were lysed, and the supernatant was used for protein quantification. Protein levels in different groups were expressed as a ratio to that of the corresponding β-actin or GAPDH. We used the following primary antibodies: rabbit anti-IL-1β, rabbit anti-TNF-α, mouse anti-IL-6 (all from Abcam, Cambridge, MA, USA), rabbit polyclonal anti-Cyr61 (Invitrogen, Carlsbad, CA, USA), rabbit anti-phospho-NF-κB p65, rabbit anti-NF-κB p65, rabbit anti-GAPDH (all from Cell Signaling Technologies, Beverly, MA, USA), and rabbit anti-β-actin (BioTNT, Shanghai, China). The secondary antibodies used were goat anti-rabbit IgG HRP, goat anti-mouse IgG HRP, and donkey anti-goat IgG HRP (all from Beyotime Biotechnology, Shanghai, China). The target proteins were examined using a Tanon 5200 imaging system (Tanon Science & Technology Corporation, Shanghai, China) and visualized with an autoradiography film.