Collagen 2 monoclonal antibody

In the light of a previous delineation, the expression of Collagen-II was observed through ICC [30 (link)]. The treated HNPCs were collected and washed with PBS, and then fixed with 4% paraformaldehyde at room temperature for 15 min. Next, cells were treated with 0.2% of Triton X-100 (P0096, Beyotime, Shanghai, China) for 10 min, and incubated firstly with 10% goat serum (C0265, Beyotime, Shanghai, China) for 20 min, and then with the diluted primary antibody (Collagen II Monoclonal Antibody; MA1-37,493, Thermo Fisher Scientific, Waltham, Massachusetts, USA) at 4°C overnight. Subsequently, cells were incubated with the diluted secondary antibody (Rabbit Anti-Mouse IgG H&L; ab6728, Abcam, Cambridge, UK) at room temperature for 1 h. Later, the cells were treated with DAB working solution (P0202, Beyotime, Shanghai, China), and then washed with distilled water. Lastly, the Collagen-II content of cells was observed (under 200 × magnification) using fluorescence microscope (N-STORM, Nikon, Tokyo, Japan).

RHu PDGF-BB, IL-1β and imatinib mesylate were purchased from MedChemExpress, United States (cat. nos. HY-P7055; HY-P7097; HY-50946); PDGF-BB polyclonal antibody, collagen II monoclonal antibody and aggrecan neo polyclonal antibody were purchased from Thermo Fisher Scientific, United Kingdom (cat. nos. PA5-88272; MA5-12789; PA1-1746); MMP3 antibody, MMP9 antibody, Adamts4 antibody, Adamts5 antibody and Caspase1 antibody were purchased from Abcam, United Kingdom (cat. nos. ab52915; ab76003; ab185722; ab41037; ab179515); PDGFR antibody, P-PDGFR antibody, P-P38 antibody and P-JNK antibody were purchased from Hangzhou Huaan Biotechnology Co., Ltd., China (cat. nos. SN0646; R1510-44; ER2001-52; ET1601-28); P-PI3K antibody, PI3K antibody, P-AKT antibody, AKT antibody, P38 antibody, JNK antibody, P-ERK antibody, ERK antibody, β-actin antibody and NLRP3 antibody were purchased from Cell Signaling Technology, United States (cat. nos.17366; 4292; 9271; 4691; 9212S; 9252S; 4370; 4695; 4970; ab263899); IL-1β antibody was purchased from ABclonal Technology Co., Ltd., China (cat. no. A1112).