Ab122897

Antibodies against GRP78 (ab21685), ATF6 (ab122897), p-PERK (ab192591) and p-eIF2α (Ser51) (ab32157) were obtained from Abcam (Cambridge, UK). Antibodies against total PERK (sc-13073) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against p-IRE1 (S724) (CY5605), eIF2α (AB3335), and β-actin (AB0061) were purchased from Abways Technology (Shanghai, China). Antibodies against total IRE1 (3294S) were purchased from Cell Signaling Technology (Danvers, MA, USA). Thapsigargin (Tg) (Abcam, ab120286), GSK2606414 (HY-18072) (MedChem Express, Monmouth Junction, NJ, USA), APY29 (HY-17537) (MedChem Express, Monmouth Junction, NJ, USA) and 4μ8C (HY-19707) (MedChem Express, Monmouth Junction, NJ, USA) were dissolved in DMSO.

Mouse liver tissues were homogenized in RIPA buffer with protease and phosphatase inhibitors (Bimake, Houston, USA). The protein concentration was determined using a BCA Protein Quantitative Assay Kit (Shenergy Biocolor Bioscience & Technology Co., Shanghai, China). Total protein was mixed with SDS loading buffer and subjected to SDS-PAGE on a 10% gel. Proteins were electrotransferred onto PVDF membranes (Millipore) and the blots were probed with the following primary antibodies overnight at 4 °C: anti-FASN (cst3180, Cell Signaling Technology, USA), anti-SREBP1C (ab3259, Abcam, USA), anti-SCD1 (ab19862, Abcam), anti-CPT1α (ab176320, Abcam), anti-MTP (sc-135994, Santa Cruz Biotechnology, USA), anti-CD36 (18836-i-ap, Proteintech, China), anti-FGF21 (ab171941, Abcam), anti-BIP (11587-1-ap, Proteintech), anti-p-IRE (ab48187, Abcam), anti-IRE (ab37073, Abcam), anti-eIF2α (cst9722, Cell Signaling Technology), anti-p-eIF2α (cst3597, Cell Signaling Technology), anti-ATF4 (10835-1-ap, Proteintech), anti-ATF6 (ab122897, Abcam), anti-p-PERK (sc-32577, Santa Cruz Biotechnology), anti-PERK (ab65142, Abcam) and anti-GAPDH (60,004–1, Proteintech). Appropriate secondary antibodies conjugated to horseradish peroxidase (Amersham) were diluted 1:5000 used. The bound primary antibodies were visualized using the Alpha Q detection system.