Precisionglide 25g needle

Transfected Huh-7 cells were washed with ice-cold PBS and scraped from the wells in ice-cold buffer K (1 mM Tris–HCl, 1 mM EGTA, and 1 mM MgCl₂, pH 7.6) containing protease inhibitor cocktail (Sigma–Aldrich; catalog no.: P2714). Cells were manually lysed through 20 passes through a BD PrecisionGlide 25G needle, and the total protein concentrations were measured with a Coomassie (Bradford) protein assay kit (Thermo Fisher Scientific; catalog no.: 23200). Proteins (25 μg) were resolved by SDS-PAGE (10%). A polyclonal rabbit primary antibody to human MTP (Abcam; catalog no.: ab63467) and a monoclonal rabbit antibody to β-actin (Cell Signaling Technology; catalog no.: 8457) were used at 1:1000 dilution. Anti-rabbit immunogobulin G, horseradish peroxidase–linked secondary antibody (Cell Signaling Technology; catalog no.: 7074) was used at 1:2000 dilution. The blots were developed with a ChemiDoc Touch Imaging system (Bio-Rad). To determine the MTP activity, 50 μg of total proteins was used. Fluorescently labeled triglyceride transfer assays were performed as previously described (25 (link), 28 (link), 29 (link), 47 (link)).

Elvitegravir (EVG, E509000) was obtained from Toronto Research Chemicals, Inc. (Ontario, Canada). Sterile phosphate-buffered saline (PBS) (10100- 031) was sourced from Gibco (Dublin, Ireland). LC/MS-grade acetonitrile (A955) and formic acid (AC270480010), BD PrecisionGlide 25G needle (14-826-49), and BD 1 Ml TB syringe (14-826-88) were procured from Fisher Scientific (Hampton, NH, USA).