Pvdf membranes and protein standards

Antibodies for p-PKR (Thr451, Millipore), PKR, eIF2α, p-eIF2α
(Ser51), p-PERK (Thr980), PERK, CHOP, caspase 3, PKR and IR were from Santa Cruz
Biotechnology (Santa Cruz, CA). The well-characterized primary antibody for NOX4 was a
kind gift from Dr. A. Shah (King's College London British Heart Foundation Centre,
London)^{21 (link)}, p-JNK (Thr183/Tyr185),
JNK, p-AKT (Ser473) and AKT were obtained from Cell Signaling Technology (Danvers, MA).
Antibodies for p-IRE1α (Ser724) and PTP1B were purchased from Abcam (Cambridge,
MA). Antibodies for p-IRS1 (Tyr608) and IRS1 were from Millipore Corp. (Billerica, MA).
Antibodies for pIR (Tyr1162/Tyr1163) were from Invitrogen (Carlsbad, CA). PVDF membranes
and protein standards were obtained from BIO-RAD (Hercules, CA). The enhanced
chemiluminescent (ECL) detection reagents were from Thermo Fisher Scientific Inc.
(Piscataway, NJ). The antibodies used are listed in Table
1.

Cholesterol, HDL cholesterol, and triglyceride concentrations were determined using kits purchased from GTLab (Buenos Aires, Argentina). Antibodies for eIF2α, p-eIF2α (Ser51), ERK, p-ERK (Thr202/Tyr204), JNK, p-JNK (Thr183/Tyr185), NOX2, NOX4, p47phox, β-tubulin, MCP-1, TNFα, p-PERK (Thr980), PERK, PTP1B, sXBP1, cATF6, IRE1α, p-IR (Tyr1162/Tyr1163), and IR were from Santa Cruz Biotechnology (Santa Cruz, CA). Primary antibodies for p-PKCδ (Thr505), p65, p-p65, IKβα, p-IκBα (Ser32), IKKα, p-IKKα/β (Ser178/180), p-AKT (Ser473), AKT, and were obtained from Cell Signaling Technology (Danvers, MA). Antibody for p-IRE1α (Ser724) was purchased from Abcam (Cambridge, MA). Antibodies for p-IRS1 (Tyr608) and IRS1 were from Millipore Corp. (Billerica, MA). Catalogue numbers for all antibodies are included in supplemental table 1. PVDF membranes and protein standards were obtained from BIO-RAD (Hercules, CA). The ECL Western blotting system was from Thermo Fisher Scientific Inc. (Piscataway, NJ). Fructose was purchased from Saporiti Labs (Buenos Aires, Argentina). EC and all other reagents were from the highest quality available and were purchased from Sigma (St. Louis, MO).

Cholesterol and triglyceride (TG) concentrations were determined using kits purchased from Wiener Lab Group (Rosario, Argentina). Free fatty acids (FFA) and insulin levels were determined using kits from Coat-A-Count (Siemens, CA). Antibodies for α-tubulin (sc-23948), and the insulin receptor (IR)β (sc-711) were from Santa Cruz Biotechnology (Santa Cruz, CA). Primary antibodies for extracellular signal-regulated kinases 1/2 (ERK1/2) (#4695), p-ERK (Thr202/Tyr204) (#4370), JNK (#9252), p-JNK (Thr183/Tyr185) (#9251), p-PKCδ (Thr505) (#9374), IKKα (#2682), p-IKKα/β (#2697) (Ser176/180), protein kinase B (AKT) (#4691), p-AKT (Ser473) (#4060), were obtained from Cell Signaling Technology (Danvers, MA). Antibody for p-IR (Tyr1162/Tyr1163) (448046G) was obtained from Invitrogen (Waltham, MA). Antibodies for insulin receptor substrate 1 (IRS1) (06–248), p-IRS1 (Tyr612) (09–432) and PTP1B (ABS40), and Luminata™ HRP chemiluminescence detection reagent were from Millipore Corp. (Billerica, MA). PVDF membranes and protein standards were obtained from BIO-RAD (Hercules, CA). The ECL Western blotting system was from Thermo Fisher Scientific Inc. (Piscataway, NJ). EC and all other reagents were purchased from Sigma (St. Louis, MO).