Txnip

15μg of total protein lysate was loaded per lane. Blots were incubated overnight with the following antibodies: Beta-actin (1:1000; EMD Millipore, MO cat #MAB1501R), Proinsulin (1:1000; CST, MA, cat #3014S), TXNIP (1:1000; MBL International, MA, cat #K0205-3), sXBP1 (1:1000; Santa Cruz Biotechnology, TX, cat #sc-7160), BiP (1:1000, CST cat #3183), Prohormone convertase 1/3 (1:1000; CST, MA, cat #11914), Prohormone convertase 2 (1:1000; CST, MA, cat #14013T), SERCA2b (1:1000, Santa Cruz Biotechnology, TX, cat #SC-8095). Blots were washed and probed with RDye infrared fluorescent dye-labeled secondary antibody conjugates (1:10,000; LI-COR biotechnology). Fluorescence intensity was determined by image studio Lite (LI-COR biotechnology). Raw uncropped western blot images can be found at https://figshare.com/search?q=10.6084%2Fm9.figshare.17161070.

Total protein samples were separated using 10–12% SDS-polyacrylamide gels and then incubated with the following primary antibodies: NLRP3 (goat polyclonal; Santa Cruz Biotechnology), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC, rabbit monoclonal; Adipogen International Inc., San Diego, CA, USA), caspase-1 (rabbit polyclonal; Santa Cruz Biotechnology), IL-1β (rabbit polyclonal; Abcam, Cambridge, UK), TXNIP (goat monoclonal; MBL International), vascular endothelial growth factor (VEGF, goat polyclonal; Santa Cruz Biotechnology), BAX (Cell Signaling Technology, Beverly, MA, USA), Bcl-2 (Cell Signaling Technology), and β-tubulin (rabbit polyclonal; Bioworld Technology Inc., St. Louis Park, MN, USA). The blots were washed with Tris-buffered saline containing Tween 20, and signals were developed using enhanced chemiluminescence (Western Chemiluminescent HRP Substrate; EMD Millipore, Billerica, MA, USA).

FMK and BI-D1870 were purchased from Axon Medchem (Groningen, The Netherlands). PP2 was purchased from Cayman Chemical (Ann Arbor, MI, USA). PF-4708671 was purchased from Selleck Chemicals (Houston, TX, USA). 2′,7′-Dichlorodihydrofluorescein diacetate (H₂DCFDA) and DAPI were purchased from Invitrogen (Carlsbad, CA, USA). TM, D-(+)-glucose and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies were purchased from the following vendors: TXNIP (MBL International, Woburn, MA, USA); ChREBP (Novus Biologicals, Centennial, CO, USA); KDEL (GRP94, GRP78) (Enzo Life Sciences, Lörrach, Germany); CREB-2 (ATF4), GADD153 (CHOP) and Src (Santa Cruz, Santa Cruz, CA, USA); Akt, p-Akt, PARP, cleaved Caspase-3, p-Src, p-S6, S6, ACC, and FAS (Cell Signaling Technology; Danvers, MA, USA); Lamin B₁ (Invitrogen); α-tubulin (Sigma-Aldrich).