Blood cellular mutant LXR-α protein stability governs initiation of coronary heart disease

Figure 2 Expression analysis of various genes upon exposure to Vitamin D₃ and Cisplatin in peripheral blood mononuclear cells of coronary heart diseasesubjects and their healthy counterparts. A: Protein expression of liver X receptor-α (LXR-α) within peripheral blood mononuclear cells (PBMCs), isolated from coronary heart disease (CHD) subjects (GS > 30) as well as normal healthy controls, exposed to culture medium enriched with and without Vitamin D₃ (1 μmol/L); B: Relative mRNA expression (change in ΔCt values) of [breast and ovarian cancer susceptibility 1 (BRCA1)-associated RING domain 1] (BARD1), BRCA1 and ATP-binding cassette A1 (ABCA1) upon Vitamin D₃ exposure in normal and patient cells; C: Protein expression of LXR-α within PBMCs, isolated from CHD subjects (GS > 30) as well as normal healthy controls, exposed to culture medium enriched with and without Cisplatin (30 μmol/L); D: Relative mRNA expression (change in ΔCt values) of ABCA1 upon Cisplatin exposure in normal and patient cells. Each bar represents mean ± SD for the combined results of three independent experiments from different individuals in triplicate. The means were compared with one way ANOVA and ^aP < 0.05 vs control group (A-D); E: Total cell lysates from PBMCs derived from CHD subjects and normal healthy individuals were immunoprecipitated with anti LXR-α antibody and immunoblotted with anti-BARD1 antibody. The direct western blotting shows the expression of LXR-α, BARD1, BRCA1 and β-actin (Sigma Aldrich) in PBMCs of CHD subjects and normal healthy controls. The experiments were repeated three times from different individuals and representative results are shown. IB: Immunoblotting; IP: Immunoprecipitation; NC: Normal cells; NS: Non-significant; PC: Patient cells.