25 hydroxycholesterol

APAP was purchased from Sigma-Aldrich (St. Louis, MO, USA). 25-Hydroxycholesterol was commercially sourced from Steraloids Inc. (Newport, RI, USA). 25HC3S was synthesized and purified in our laboratory as previously described [22 (link)]. The reagents for real-time RT-PCR were obtained from Applied Biosystems (Applied Biosystems, Foster City, CA, USA). The RT² Profiler PCR Array-Cell Death Pathway Finder was acquired from QIAGEN (Valencia, CA, USA). MitoProbe JC-1 Assay Kit for Flow Cytometry and H2DCFDA were purchased from Life Technologies (Carlsbad, CA, USA). Propylene glycol (PG), hydroxypropyl-β-cyclodextrin (HBC), paraformaldehyde, phosphate buffer, 2-thiobarbituric acid (TBA) and malondialdehyde (MDA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell culture media were obtained from Invitrogen (Carlsbad, CA, USA). All other reagents were from Sigma-Aldrich unless otherwise indicated.

We obtained MG-132 from Boston Biochem (Cambridge, MA); horseradish peroxidase-conjugated donkey anti-mouse and anti-rabbit IgGs (affinity-purified) were from Jackson ImmunoResearch Laboratories (West Grove, PA); digitonin was from Calbiochem (San Diego, CA); geranylgeraniol and farnesol from Sigma–Aldrich (St. Louis, MO) and Santa Cruz Biotechnology (Dallas, TX); biotin was obtained from Sigma–Aldrich (St. Louis, MO); and 25-hydroxycholesterol from Steraloids (Newport, RI). Apomine was synthesized by the Core Medicinal Chemistry laboratory at the University of Texas Southwestern Medical Center. Other reagents, including new born calf lipoprotein-poor serum (NC-LPPS, d > 1.215 g/ml), sodium compactin, sodium mevalonate, and stock solutions of digitonin were prepared or obtained from previously described sources (Goldstein et al., 1983 (link); DeBose-Boyd et al., 1999 (link); Elsabrouty et al., 2013 (link)).

Cholesterol and 25-hydroxyCholesterol were purchased from Steraloids (Newport, RI). [1,2-³H]Cholesterol, 25-[26,27-³H]hydroxyCholesterol and [³²P]PO₄ were purchased from PerkinElmer Life Sciences (Boston, MA). [¹⁴C]-Dipalmitoyl-phosphatidylcholine was purchased from American Radiolabeled Chemicals (St. Louis, MO). Phospholipids were purchased from Avanti Polar Lipids (Alabaster, AL). Antibodies against giantin and polypeptide N-acetylgalactosaminyltransferase (GALNT) (BioLegend, San Diego CA), vimentin (Abcam, Cambridge UK) and V5 (BioRad, Raleigh NC) were used for immunofluorescence microscopy and immunoblotting. Alexa Fluor 488- and Alexa Fluor 594-conjugated secondary antibodies (Molecular Probes, Eugene OR) and IRDye 680LT- and IRDye 800CW-conjugated secondary antibodies (LI-COR Biosciences, Lincoln NE) were used for immunofluorescence and immunoblotting, respectively.