Mcc950

4-Hydroxytamoxifen was purchased from Aladdin (Shanghai, China). Jasplakinolide, taxol and caffeine were obtained from Abcam (Cambridge, UK). 2-Aminoethyl diphenylborinate, dantrolene, Z-VAD-FMK, SP600125, lipopolysaccharide (LPS) and nigericin were purchased from MedChemExpress (Monmouth Junction, NJ, USA). MCC950 were obtained from CSNpharm (Chicago, IL, USA). PEG8000 and PF431396 were obtained from Beyotime Biotechnology (Shanghai, China). Cantharidin was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Rabbit anti-caspase-1, rabbit anti-ASC and rabbit anti-NLRP3 antibodies were obtained from Proteintech (Chicago, IL, USA). Mouse anti-α-tubulin antibody was from Boster (BM1452, Wuhan, China). Mouse anti-β-actin was purchased from Cell Signaling Technology (Danvers, MA, USA). The siRNA targeting ASC and CASPASE-1 was constructed by GenePhrama (Shanghai, China). The Flag-Gsdmd-NT, Flag-Gsdmd-CT plasmid were purchased from Addgene (Watertown, MA, USA).

Sixty male SD rats underwent lens isolation and organ culturing as previously described [42 (link), 43 (link)]. Briefly, the rats were humanely killed with a carbon dioxide (CO₂) euthanasia apparatus. Their eyes were dissected posteriorly, and lenses were extracted and cultured in M-199 medium (M5017, Sigma-Aldrich, St. Louis, USA) containing 5% fetal bovine serum (FBS, Gibco, Grand Island USA) and antibiotics (100 U/mL penicillin, 100 μg/mL streptomycin, 0.25 μg/mL amphotericin B). The cultured lenses were treated with increasing doses (200 to 1000 μM) of soluble UA (U2625, Sigma-Aldrich) to determine the effective dose for inducing cataract formation. Subsequent experiments used the established model to test the effects of 400 μM MCC950 (CSN18163, CSNpharm, Chicago, USA), a selective NLRP3 antagonist [44 (link)]. The lenses were cultured at 37 °C in 5% CO₂ for a week with daily lens opacity assessments (Supplementary Table S1 and Fig. S1). “Clear lens survival” was defined as an opacity grade ≤1 under microscopy. Additionally, lenses were collected at 12, 24, or 48 h to measure NLRP3/caspase-1/IL-1β signaling. Eventually, all lenses underwent Western blot and histological analysis.