Lomitapide

The following antibodies were used for immunofluorescence staining or western blot analyses: anti-FoxA2 (used at 1:400, Cell Signaling), anti-HNF4α (used at 1:500, Cell Signaling), anti-AFP (used at 1:1000, Sigma-Aldrich), anti-ALB (used at 1:1000, Cedarlane, Burlington, Canada), anti-ZO-1 (used at 1:1000, Thermo Fisher), anti-E cadherin (used at 1:500, Cell Signaling), anti-SR-BI (used at 1:100, Novus Biologicals), anti-BCRP (used at 1:100, Millipore), anti-MRP2 EAG5^{48 (link)} (used at 1:200, a kind gift from Anne Nies, IKP Stuttgart^{48 (link)}), anti-Apo-CIII (used at 1:500, Abcam), anti-CYP8B1 (used at 1:100, Abcam), anti-ORF2 (used at 1:400, a kind gift from Suzanne U. Emerson, NIH) and anti-HAV capsid (used at 1:1000, a kind gift from Stanley M. Lemon, UNC School of Medicine). Alexa Fluor 488 and 549 anti-mouse (used at 1:1000) and Alexa Fluor 488 and 549 anti-rabbit (used at 1:1000) antibodies were purchased from ThermoFisher. Alexa 594-conjugated transferrin was purchased from ThermoFisher. Tenofovir, Tenofovir disoproxil fumarate, and Emtricitabine were obtained through the AIDS Reagent Program, Division of AIDS, NIAID, NIH. Elvitegravir and Cobicistat were purchased from SelleckChem. Sofosbuvir was purchased from Acme Bioscience. BX795, oleic acid and lomitapide were purchased from Sigma-Aldrich.

PC (no. 131601C) and fluorescently labeled ammonium salts of PEs with the acyl groups, NBD-1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE; no. 810143C), NBD-POPE (no. 810118P), or NBD-DOPE (no. 810145C) were obtained from Avanti Polar Lipids (Alabaster, AL). NBD-1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (DPPE; no. N360) was obtained from Invitrogen (Thermo Fisher Scientific). Lomitapide, an MTP inhibitor, was obtained from Sigma (no. SML1385-5MG). The Cos-7 and Huh-7 cell lines, and the plasmids pcDNA3 and pcDNA3-hMTP-FLAG, have been described previously (20 (link), 21 (link), 26 (link), 27 (link)). Anti-FLAG® M2 affinity agarose gel and other chemicals were purchased from Millipore Sigma (no. A2220-5ML, St. Louis, MO). Round bottom black 96-well assay plates were obtained from Costar (no. 3792; Kennebunk, ME). FLAG peptide was custom synthesized (GenScript).

High-throughput cancer cell viability assays were performed by Reaction Biology Corp. (USA). About 120 major cancer cell lines derived from skin, breast, brain, ovary, liver, stomach, kidney, bone, pancreas, intestine, lung, and blood cancer were inoculated in a 96-well plate at a density of 10⁴ cells/well, and cultured at 37 °C for 24 h, followed by various concentrations (0, 1, 2, 5, 10 µM) of lomitapide (Sigma-Aldrich). The plate was incubated at 5% CO₂ at 37 °C for 24 h after treatment with lomitapide. Thereafter, cells were added to each well of 100 µl of the assay reagent (CellTiter-Glo^® Reagent), and luminescence was measured using a VICTOR X Multilabel Reader (PerkinElmer, USA).