Actin sc 47778

β-Elemene (molecular formula C15H24), whose molecular weight is 204.35, was provided by Jingang Pharmaceuticals (#081152, Dalian, China). The antibody of HIF-1α (#3716 S), LDHA (2012S), was obtained from Cell Signalling Technology (Danvers, MA, USA). The antibody GLUT1 (2646S) was purchased from NOVUS (USA), and the antibody PFKL (55028-1-AP) was purchased from Proteintech. Anti-PDK1 (ab226963) and Anti-PHD1 (ab113077) were purchased from Abcam. Anti-Ki67 antibody was purchased from Fuzhou Maixin Biological Technology (Fujian, China). ALDH3A1 (sc-137168), Cyclin B1 (166757), Cyclin D1 (8396), Cyclin E (sc-377100), and ACTIN (sc-47778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). 2-Deoxy-D-glucose (2-DG) was purchased from MedChemExpress (HY-13966, USA). The Cell Counting Kit-8 (CCK-8) was purchased from APExBIO (Catalog No. K1018, USA).

The following commercial antibodies were used: Huntingtin (MAB2166, 1:3000) and puromycin (MABE343, 1:10,000) antibodies were obtained from, Millipore-Sigma. Anti-polyglutamine (poly-Q) antibody (P1874, 1:5000) was from Sigma. Actin (sc47778, 1:20,000) and GST-horseradish peroxidase (HRP, sc138 HRP, 1:10,000) antibodies were from Santa Cruz Biotechnology. RPL7 (IHC-00455, 1:10,000), RPL35A (A305-106A, 1:10,000) and Caprin1 (A303-881A, 1:1000) from Bethyl Laboratories. mTOR (2972, 1:3000), FMRP (4317, 1:1500), and S6 (2217, 1:10,000), and normal mouse IgG (5415, 1:2500) were from Cell Signaling Technology. Mfsd (10 11518-1-527 AP, 1:1000), Acan (13880-1-AP, 1:1000), Ppbp (13313-1-AP, 1:1000), Mgp (10734-1-AP, 1:1000), and Phf11d (10898-1-AP, 1:1000) were from Proteintech. HRP-conjugated secondary antibodies were from Jackson ImmunoResearch Inc. HRP-conjugated secondary antibodies (115-035-146 (goat anti-mouse), 1:10,000; or 111-035-144 (goat anti-rabbit), 1:10,000) were from Jackson ImmunoResearch Inc. For Immunostaining Huntingtin (1:100, MAB2166), Rpl7 (1:50; IHC-00455) were used. Secondary antibodies for STED microscopy were anti-mouse STAR 635p (1:400) and anti-rabbit Alexa 594 (1:400) that were self-coupled in-house and the company that produced them is Abberior, Göttingen, Germany.

DMEM, FBS, goat anti-mouse IgG secondary antibody conjugated to Alexa Fluor 488, Fura-2 AM and all RT-qPCR reagents were obtained from Invitrogen (Carlsbad, CA). Neurotensin, insulin and phalloidin-TRITC were obtained from Sigma Chemical (St. Louis, MO). The geranylgeranyl transferase I (GGTase I) inhibitor GGTI 298, PI 3-Kinase inhibitor A66 and the MEK inhibitor PD0325901 were from R&D Systems (Minneapolis, MN). The dual PI3K/mTOR inhibitor NPV-BEZ235 was purchasedfrom Selleck Chemicals (Houston, TX). Primary antibodies used were as follows: YAP (H-9, sc-271134 and 63.1, sc-101199, final dilution 1:200 for immunofluorescence), tubulin (sc-5274; final dilution 1:400), actin (sc-47778; final dilution 1:400) and GAPDH (sc-365062; final dilution 1:400) (Santa Cruz Biotechnology); phospho-YAP Ser127 (D9W2I, 13008; final dilution 1:1000), YAP (15028; final dilution 1:1000 for western blots), phospho-p70 S6 KinaseThr389) (9205; final dilution 1:1000) and phospho-S6 Ribosomal Protein Ser240/244 (5364; final dilution 1:1000), phospho LATS Thr1079 (8654; final dilution 1:1000) and LATS2 (5888; final dilution 1:1000) were from Cell Signaling Technology (Danvers, MA). Horseradish peroxidase–conjugated anti-rabbit IgG and anti-mouse IgG were from GE Healthcare Bio-Sciences Corp (Piscataway, NJ). All other reagents were of the highest grade available.

This study used reagents obtained from the commercial companies indicated below. Fetal bovine serum (FBS, Cat No. 12483-020) and TRIzol (GIBCO BRL, Gaithersburg, MD, USA); RPMI-1640 medium, Tween-20, bovine serum albumin (BSA), skim milk, and rapamycin (Sigma-Aldrich Chemical Co., St. Louis, MO, USA). Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA); Abs against molecules such as LAMP-2 (sc-18822), lamin B (sc-374015), PUMA (sc-377015), and actin (sc-47778) (Santa Cruz Biotechnology, Santa Cruz, CA, USA); Abs against molecules such as ATG5 (Cat No. 9164), ATG12 (Cat No. 4180), FoxO3a (Cat No. 12829), AMPKα (Cat No. 5832), phospho-AMPKα (Cat No. 2535), and Bax (Cat No. 2772), and horseradish peroxidase-conjugated anti-rabbit IgG (Cat No. 7074) and anti-mouse IgG (Cat No. 7076) (Cell Signaling Technology, Inc., Beverly, MA, USA); anti-LC3 antibody (Cat No. NB100-2220, Novus Biologicals, Saint Charles, MO, USA); p62/SQSTM1 (Cat No. H00008878-M01, Abnova, Taipei City, Taiwan); DyLight 549- and Alexa Fluor 488-conjugated secondary Abs (Thermo Fisher Scientific, Waltham, MA, USA); Vectashield mounting medium with 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories, Inc., Burlingame, CA, USA); compound C (HY-13418A, Medchemexpres, Monmouth, NJ, USA); bafilomycin A1 (Tocris Bioscience, Bristol, UK); 3-MA (Merckmillipore, Darmstadt, Germany)

Human or mouse liver tissues or primary mouse hepatocytes were lysed using RIPA buffer (0.5 M Tris-HCl, pH 7.4, 1.5 M NaCl, 2.5% deoxycholic acid, 10% NP-40, and 10 mM EDTA) with protease inhibitors (MilliporeSigma). The Pierce BCA Protein Assay Kit (Thermo) was used to measure the concentration of protein. The standard procedures of SDS-PAGE were performed using 30 μg lysates and transferred to nitrocellulose membranes. The SuperSignal West Pico Chemiluminescent Substrate (Thermo) was used according to the manufacturer’s protocol. The following Abs were used: CYP4A (sc-271983; Santa Cruz Biotechnology), ACTIN (sc-47778; Santa Cruz Biotechnology), and CYP2E1 (AB1252; Millipore).