Cst3597

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.

The following antibodies were used for immunoblot analysis: rabbit-anti-IKK2 (CST-2678, Cell Signaling, Danvers, MA, USA), rabbit-anti-p-p65 (CST-3033, Cell Signaling, Danvers, MA, USA), rabbit-anti-p65 (sc-372, Dallas, TX, USA), chicken-anti-GFP (ab13970, Cambridge, UK), rabbit-anti-tubulin (ab6046, Cambridge, UK), rabbit-anti-PLP1 (CST-28702, Cell Signaling, Danvers, MA, USA), rabbit-anti-MOG (CST-96457, Cell Signaling, Danvers, MA, USA), rabbit-anti-MBP (CST-2396, Cell Signaling, Danvers, MA, USA), rabbit-anti-GAPDH (CST-3686, Cell Signaling, Danvers, MA, USA), rabbit-anti-p-eIF2α (CST-3597, Cell Signaling, Danvers, MA, USA), rabbit-anti-eIF2α (CST-5324, Cell Signaling, Danvers, MA, USA), rabbit-anti-ERK2 (sc-1647, Dallas, TX, USA).
For immunofluorescence, the following primary antibodies were used: mouse-anti-CC1 (OP80, Merck, Darmstadt, Germany), mouse-anti-GSTπ (BD610719, BD, Franklin Lakes, NJ, USA), chicken-anti-GFP (GFP-1020, AvesLab, Davis, CA, USA), mouse-anti-GFAP (sc-33673, Dallas, TX, USA), rabbit-anti-NG2 (AB5320, Merck, Darmstadt, Germany), rabbit-anti-MBP (Biolegend 836,504, San Diego, CA, USA), mouse-anti-Neurofilament-H (SMI32P) (Biolegend 801,701, San Diego, CA, USA), rabbit-anti-ß3-tubulin (Biolegend 802,001, San Diego, CA, USA), rabbit- anti RFP (abcam ab124754, Cambridge, UK).