Effects of Con-miR and Con-Anti-miR on intracellular cholesterol accumulation, cholesterol efflux and the expression of miR-33a-5P, SREBP2, ABCA1 and ABCG1 in THP-1 macrophages.

THP-1 macrophages were incubated in serum-free medium at 37°C for 24 h, after lentivirus (Con-miR and Con-Anti-miR) infection for 48 h. (A) THP-1 macrophages examined for lipid inclusions by oil red O staining. (B) Semi-quantitative analysis of oil red O positive staining of THP-1 macrophages for the examination of lipid inclusions. Data are means ± SD from 6 separate fields. (C) Intracellular free cholesterol (FC), total cholesterol (TC) and cholesterol ester (CE). (D) ApoA-I-mediated cholesterol efflux. Data are means ± SD of duplicate wells from 6 experiments. (E) mRNA levels determined by the 2−ΔΔCt method for RT-PCR. U6 or β-actin served as a reference gene. Data are means ± SD from 6 experiments. (F) Protein levels examined by Western blotting. (G) Quantification of the densitometric values of SREBP2, ABCA1 and ABCG1 protein bands from four experiments, normalized to β-actin and expressed as a percentage of the control. Data are means ± SD.