Collagenase type 2

FKA (T3S0737, purity: 99.41%) and 3-Methyladenine (3 MA) were acquired from TOPSCIENCE (Shanghai, China), and the mouse IL-1β cytokine was obtained from the R&D system (501-RL-010, United States). Safranin O solution were provided by Solarbio (Beijing, China). The primary antibodies for iNOS, COX2, Col2, Aggrecan, Sox9, and immunofluorescence secondary antibodies were acquired from Abcam (Shanghai, China). Atg12, Beclin, LC3I/II, P21, P16, Bax, Bcl2, GAPDH, and antibodies for all pathways were purchased from CST (Beverly, MA, United States). Corresponding primary antibodies for MMP3/13 and Nrf2/HO-1/NQO1 were supplied by the Proteintech Group (Wuhan, Hubei, China). ADAMTS5 primary antibody, secondary antibody, phosphate-buffered saline (PBS), trypsin, collagenase type II, the CCK8 assay kit, bovine serum albumin (BSA), and protein extraction kit were ordered and acquired from Boster Biological Technology (Wuhan, Hubei, China). RFP-GFP-LC3-adenovirus was purchased from Hanbio (Shanghai, China). The Senescence β-galactosidase staining kit and the Annexin V-FITC apoptosis detection kit were supplied by Beyotime (Shanghai, China).

Alantolactone (HY-N0038) was obtained from MedChem Express (MCE). Recombinant mouse IL-1β cytokine was provided from R&D Systems (501-RL-010, United States). Solarbio supplied safranin O liquor (Beijing, China). Primary antibodies applied in this research were: anti-iNOS, anti-MMP-13 which were acquired from Abcam (Shanghai, China), anti-COX2, anti-LC3Ⅱ/Ⅰ, anti-P-STAT3, anti-P-P65, anti-P-IκB, anti-P/T-mTOR which were purchased from CST (Beverly, MA, United States), anti-STAT3, anti-MMP-1, anti-MMP-3, anti-ATG5, anti-P62, anti-P65, anti-IκB, anti-P/T-PI3K, anti-P/T-AKT, anti-GAPDH which were afforded from Proteintech Group (Wuhan, Hubei, China) and anti-ADAMTS5 which was supplied from Boster Biological Technology (Wuhan, Hubei, China). Secondary antibodies, Cy3 and FITC Conjugated AffiniPure Goat Anti-Rabbit IgG, DAPI staining solution, collagenase type II and trypsin were got from Boster Biological Technology (Wuhan, Hubei, China). HanBio Inc. (Shanghai, China) served the mRFP-GFP-LC3 adenoviral vectors.

LRRK2-IN-1 (CAS 1234480-84-2) was purchased from Topscience (T2246; China). Recombinant mouse IL-1β cytokine (#501-RL-010) and human IL-1β cytokine (201-LB-025) were obtained from R&D Systems (USA). The primary antibodies against glyceraldehyde 3-phosphate dehydrogenase (GAPDH; #60004-1-Ig), inducible nitric oxide synthase (iNOS; #22226-1-AP), cyclooxygenase 2 (COX2; #66351-1-Ig), type II collagen (COL2; #28459-1-AP), aggrecan (#13880-1-AP), and matrix metallopeptidase 13 (MMP13; 18165-1-AP) were acquired from Proteintech Group (China) and the primary antibodies against matrix metallopeptidase 3 (MMP3; # BM4074) and SRY-box transcription factor (SOX9; #A00177-2) from Boster Biological Technology (China). Trypsin, collagenase type II, phosphate buffer saline (PBS) buffer solution, and the secondary antibodies for western blot and immunofluorescence analyses were provided by Boster Biological Technology (China). In addition, fetal bovine serum (FBS) and Dulbecco’s modified Eagleʼs medium F12 (DMEM/F12) were provided by Gibco (USA). Safranin O solution was provided by Solarbio (#G1067; China).