2 oleoyl 1 palmitoyl sn glycero 3 phosphocholine

Reconstituted HDL (rHDL) particles containing apoA-I, phosphatidylcholine, and cholesterol were prepared using 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (Sigma-Aldrich, Germany), cholesterol (Sigma-Aldrich), recombinant human apoA-I (produced as described (41 )), and sodium cholate in a molar ratio 100:10:1:100 in 10 mM Tris-HCl buffer, pH 8, containing 0.01% EDTA, 150 mM NaCl (41 ).

Lipids, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and 1,2dimyristoyl-sn-glycero-3-phosphocholine (DMPC), were purchased from Sigma (St. Louis, MO, USA) and used without further purification.

1-palmitoyl,2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), chicken egg yolk sphingomyelin (SM), and cholesterol (Chol) were purchased from Sigma-Aldrich (Darmstadt, Germany). The lipids were dissolved in chloroform and divided into 1 mg aliquots. Then, the organic solvent was evaporated under nitrogen stream and kept at −20 °C.
Phosphate buffered saline (PBS) pH 7.4 (137 mM NaCl, 2.7 mM KCl and 10 mM phosphate) was prepared from PBS tablet (Sigma-Aldrich, Darmstadt, Germany). The buffer tablet was dissolved in ultrapure water (Milli-Q, Merck, Darmstadt, Germany) and filtrated through a 0.22 µm filter (Whatman, GE Health care life science, Chicago, IL, USA).

The sodium dodecyl sulfate (SDS), lipopolysaccharides (LPS), 2-Oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC), 2-Oleoyl-1-palmitoyl-sn-glycero-3-phospho-rac-1-glycerol (POPG), and 2-Oleoyl-1-palmitoyl-sn-glycero-3-phosphoethanolamine (POPE) lipids, the resin for peptide synthesis, and protected amino acids were obtained from Sigma-Aldrich. Sheep RCB were from GRASO (Poland). All chemicals were of analytical or reagent grade and the buffers were prepared using distilled water.

Plasticware was obtained from Costar (Merck, Darmstadt, Germany) and Sarstedt (Nümbrecht, Germany). Materials for real-time quantitative PCR were obtained from Applied Biosystems (Thermo Fisher Scientific, Waltham, MA). Organic solvents for TLC, paraformaldehyde, glutaraldehyde, and Hoechst 33342, were obtained from Sigma-Aldrich (St. Louis, MO). Phalloidin-CF568 conjugate was obtained from Biotium (Fremont, CA). Bodipy 493/503 was obtained from Thermo Fisher Scientific.
Lipids used for TLC were purchased from two sources: 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (#42773), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphoethanolamine (#01991), and 2-oleoyl-1-palmitoyl-sn-glycero-3-phospho-L-serine sodium salt (#51581) were obtained from Sigma-Aldrich (St. Louis, MO). Hexadecanoic acid (palmitic, #N-16-A), octadecanoic acid (stearic, #N-18-A), 9-hexadecenoic acid (palmitoleic, #U-40-A), 9-octadecenoic acid (oleic, #U-46-A), 7-octadecenoic acid (vaccenic, #U-48A), 9-12 octadecadienoic acid (linoleic, #U-59-A), trihexadecenoin (palmitolein, #T-215), 13-docosenoic acid (erucic, #U-79-A), dipalmitolein (#D-216), trioctadecenoin (#T-235), diolein (#D-236), diolein 1-3 Isomer (#D-237), monoolein (#M-239), cholesteryl palmitate (#CH-815), cholesteryl palmitoleate (#CH-826), and cholesteryl oleate (#CH-828) were obtained from Nu-Chek Prep Inc. (Elysian, MN).

D₃, ergocalciferol (vitamin D₂), 25(OH)D₃, 1α(OH)D₃, retinyl acetate, 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (phosphatidylcholine), 1-palmitoyl-sn-glycero-3-phosphocholine (lysophosphatidylcholine), monoolein, free cholesterol, oleic acid, sodium taurocholate, and simvastatin were purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, France). Ezetimibe beta-d-glucuronide was purchased from Sequoia Research (Pangbourne, UK). Block lipid transport-1 (BLT1) was purchased from ChemBridge (San Diego, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM) containing 4.5 g/L glucose and trypsin-EDTA (500 mg/L and 200 mg/L, respectively), non-essential amino acids, penicillin/streptomycin and PBS were purchased from Life Technologies (Illkirch, France), and fetal bovine serum (FBS) came from PAA (Vélizy Villacoublay, France). ASBT tagged with the DDK peptide (DYKDDDDK flag peptide) in pCMV plasmid and DDK antibody were purchased from Origen (Austin, TX, USA). All solvents used were HPLC grade (Carlo Erba Réactifs-SdS, Val de Reuil, France).