Protein Expression Analysis in RAW264.7 Cells and Chondrocytes

Total proteins of RAW264.7 cells and primary chondrocytes were acquired by the RIPA lysis buffer (#R0010, Solarbio, China) with the supplement of 1 mM phosphatase inhibitor cocktail (#B15002, Bimake, USA) and 1 mM phenylmethanesulfonyl fluoride (#329-98-6, Solarbio). Western blot was executed as previously described [9 (link)]. The primary antibodies used in this work were: p-CaMKII (#ab124880, Abcam, USA), CaMKII (#ab134041, Abcam), iNos (#13120S, Cell Signaling Technology, USA), Cox2 (#12282S, Cell Signaling Technology), Col II (#BA0533, Boster, Wuhan, China), Mmp13 (#GB11247, Servicebio, China), Fth1 (#4393, Cell Signaling Technology), Gpx4 (#ab125066, Abcam), Cd206 (#18704-1-AP, Proteintech, China), Arg-1 (#16001-1-AP, Proteintech), and Gapdh (#5174, Cell Signaling Technology). After the incubation of horseradish peroxidase-conjugated goat anti-rabbit/mouse secondary antibodies (#BL003A or #BL001A, Biosharp), western blots were imaged by the ChemiDocXRS + Imaging System (Tanon, Shanghai, China). Quantification of proteins was performed by the ImageJ software (version 1.8.0). All primary and secondary antibodies used in this work were commercial.