Protein a g agarose sepharose beads

RIPA lysis buffer, BCA protein assay kit, Protein A/G agarose/sepharose beads, and Lactate Dehydrogenase (LDH) Cytotoxicity Assay Kit were purchased from the Beyotime Institute of Biotechnology (Shanghai, China). The following antibodies were used: anti-BIP (3177S), anti-CHOP (2895T), anti-NLRP3 (15101S), IRE1a (3294T), and p-eIF2α (3398T) were purchased from Cell Signaling Technology, Inc.; anti-GSDMD (A18281) was obtained from ABclonal Technology (Wuhan, China); anti-TXNIP (sc-166234) was bought from Santa Cruz Biotechnology, Inc.; anti-β-actin monoclonal antibody was purchased from Proteintech Group (Chicago, IL); goat anti-NLRP3 (49012) was purchased from SAB Laboratories, Inc. (Logan, UT, USA). Middlebrook 7H9 and 7H10 media were obtained from Difco (Detroit, MI, USA), and oleic acid-albumin-dextrose-catalase (OADC) supplements were from BD Biosciences (BD, Sparks, MD, USA). RiboFECT CP Transfection Kit was purchased from Ribo Biotechnology (Guangzhou, China). IL-1β ELISA Kit was purchased from R&D (USA).

RIPA lysis buffer, BCA Protein Assay Kit, and Protein A/G agarose/sepharose beads were obtained from the Beyotime Institute of Biotechnology (Shanghai, China). The following antibodies were used: anti-NLRP3, anti-IL-1β, anti-ASC, anti-LC3, anti-p62, anti-Akt, anti-phosphorylated Akt (Ser473), anti-mTOR, anti-phosphorylated mTOR (Ser2448), anti-phosphorylated p38, anti-phosphorylated JNK, and anti-phosphorylated ERK1/2 were purchased from Cell Signaling Technology, Inc. (CST, Danvers, MA, USA); anti-IL-1β was purchased from R&D Systems (Minnesota, USA); BECN1 siRNA, Transfection Reagents, rabbit anti-caspase-1, goat anti-rabbit, donkey anti-goat, and goat anti-mouse LC3 were purchased from Santa Cruz Biotechnology, Inc.; anti-β-actin monoclonal antibody was from ProteinTech Group (Chicago, IL); goat anti-NLRP3 was purchased from Abcam (Cambridge, UK). Guttiferone K (GK, purity > 98%, C38H50O6, MW: 602.80) was isolated from Garcinia yunnanensis as previously described [29 (link)]; Dulbecco's Modified Eagle's Medium (DMEM) was obtained from HyClone Laboratories, Inc. (Logan, UT, USA). Middlebrook 7H9 and 7H10 media were obtained from Difco (Detroit, MI, USA), and oleic acid-albumin-dextrose-catalase (OADC) supplements were from BD Biosciences (BD, Sparks, MD, USA).