Ac yvad cmk

Five renal cell carcinoma cell lines (786-O, ACHN, A498, CAKI-1, and OSRC-2) and a human renal tubule epithelial cell line (HK-2) were purchased from the American Type Culture Collection (Manassas, USA). 786-O and OSRC-2 cell lines were maintained in RPMI-1640 culture medium (GIBCO), ACHN and A498 cell lines were maintained in minimal essential medium with Earle’s balanced salts, and CAKI‑1 cells were cultured in McCoy’s 5A medium. All culture media were supplemented with 10% heat-inactivated FBS (FBS; GIBCO) and 1% streptomycin–penicillin. HK-2 cell line was cultured in complete medium containing K-SFM (keratinocyte serum-free medium). All cell lines were maintained in an incubator with 5% CO₂ at 37 °C. JQ1 (Catalog NO. S7110), MCC950 (Catalog NO. S7809), and BAY 11-7082 (Catalog NO. S2913), Z-DEVD-FMK (Catalog NO. S7312) were all purchased from Selleckchem. Ac-YVAD-CMK (Catalog NO. N-1330) was purchased form BaChem, and TNF-α (Catalog NO. 300-01A) was obtained from PeproTech.

RPMI 1640 and the Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Gibco. DCFH-DA probe was from Beyotime (Shanghai, China). Anti-mouse/human PPARγ (sc-7273, sc-7196, the dilution ratio was 1:500), anti-mouse caspase-1 (G-19, sc-1597, the dilution ratio was 1:1,000), MCAD (sc-98926 Lot # G2809, the dilution ratio was 1:1,000), EF-Tu (sc-393924, Lot # B0514, the dilution ratio was 1:1,000), SFXN1 (sc-160797, Lot # F0311, the dilution ratio was 1:1,000) and HA (sc-7392, Lot # E0614, the dilution ratio was 1:1,000) antibodies were purchased from Santa Cruz Biotechnology, Dallas, TX, USA. Anti-mouse FITC-F4/80 was purchased from eBiosience (Ref: 11-4801, the dilution ratio was 1:1,000). Anti-human caspase-1 antibody (D7F10, the dilution ratio was 1:1,000) was from Cell Signaling (Boston, USA). HRP conjugated goat anti-mouse/rabbit antibodies were from Beyotime (Haimen, China, the dilution ratio was 1:10,000). All cytokines such as M-CSF were obtained from Peprotech Systems (Minneapolis, MN). The pan-caspase inhibitor z-VAD-FMK, caspase-1 inhibitor z-WEHD-FMK and the caspase-3 inhibitor z-DEVD-FMK were obtained from R&D systems. The caspase-1 inhibitor Ac-YVAD-CMK was obtained from Bachem. NCX-4016 (NO-aspirin) were purchased from Nicox (Sophia Antipolis, France). All of other chemicals were purchased from Sigma-Aldrich unless otherwise described.

LPS, ATP, nigericin, poly dA:dT, staurosporine, cytochalasin D, valinomycin, and apyrase were purchased from Sigma-Aldrich (St. Louis, MO, USA). JC-1 and MitoSox were obtained from Invitrogen (San Diego, CA, USA). zVAD-FMK,and ac-YVAD-CMK were purchased from Bachem (Torrance, CA, USA). Mammalian expression constructs for mouse Sarm1 (pGW1-Myc-Sarm1) was purchased from Addgene (Watertown, MA, USA). The following antibodies were used for detecting mouse caspase-1 (Adipogen, San Diego, CA, USA), NLRP3 (Adipogen), IL-1β (R&D Systems, Minneapolis, MN, USA), ASC (Santa Cruz Biotechnology, Dallas, TX, USA), β-actin (Santa Cruz Biotechnology), caspase-3 (Cell Signaling, Beverly, MA, USA), gasdermin D (Abcam, Cambridge, MA, USA), and Sarm1 (Cell Signaling).