Recombinant human globular adiponectin gacrp

Recombinant human globular adiponectin (gAcrp) was obtained from Peprotech Inc. (Rocky Hill, NJ, USA). LPS, adenosine 5′-triphosphate (ATP), and 3-methyadenine (3-MA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). A Membrane Permeability/Dead Cell Apoptosis Kit with PO-PRO-1 and 7-Aminoactinomycin D for Flow Cytometry were purchased from Molecular Probes (Eugene, OR, USA). A Caspase-1/ICE Colorimetric Assay Kit was obtained from Biovision (Mountain View, CA, USA), and 5-aminoimidazole-4-carboxamide-1-b-4-ribofuranoside (AICAR), a pharmacological activator of AMPK, was obtained from Calbiochem (San Diego, CA, USA). Primary antibodies against phosphorylated and total AMPKα (phospho-specific and total form), IL-1β, and LC3 were obtained from Cell Signaling Technology Inc. (Beverly, MA, USA), β-actin was obtained from Thermo Scientific (Hudson, NH, USA), and NLRP3 was purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against caspase-1 and ASC were purchased from Adipogen (San Diego, CA, USA). Secondary goat-anti-rabbit and anti-mouse antibodies were obtained from Thermo Scientific.

All cell culture reagents used in this study were obtained from HyClone Laboratories (South Logan, UT, USA). Recombinant human globular adiponectin (gAcrp) was purchased from Peprotech Inc. (Rocky Hill, NJ, USA). Bafilomycin A1, an inhibitor of autophagosome-lysosome fusion, was ordered from Sigma-Aldrich (St. Louis, MO, USA). Antibodies targeting p62 (Catalog. No. 5114), phospho-specific p38 MAPK (Catalog. No. 9215), total p38 MAPK (Catalog. No. 9212), LC3II (Catalog. No. 2775), Beclin-1(Catalog. No. 3738), and Nrf2 (Catalog. No. 8882) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Antibodies against TRAF6 (Catalog. No. Sc-7221), p21 (Catalog. No. sc-397) and ubiquitin (Catalog. No. sc-8017) were taken from Santa Cruz (Delaware, CA). Antibodies against Atg5 (Catalog. No. PAI-46178), β-actin (Catalog. No. 04-1116) and chemiluminescent substrate (product no. NCI4080KR) were from Thermo Scientific Inc. (Rockford, IL, USA). The TNF-α ELISA kits were purchased from BioLegend (San Diego, CA, USA). Until and unless mentioned elsewhere, all other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).