Fig. 2

AZ7235 increases ABCA1 expression. (a) Cellular ABCA1 protein levels were measured by immunoblot in CCF-STTG1 cells after 72 h treatment with DMSO, 1 μM T0901317, or 3 μM AZ7235. Representative blot was generated from same gel shown in Fig. 1f. (b) CCF-STTG1 cells were labeled with ³H-cholesterol with co-treatment of DMSO alone, 1 μM T0901317, or 3 μM AZ7235 for 24 h. Cholesterol efflux over 24 h in the absence (NA) or presence of 10 μg/ml of lipid-free apoA-I along with the above drug treatment was evaluated. Graphs represent mean % efflux and standard deviation of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001 comparing drug effect over respective DMSO control; ### P < 0.001 comparing between NA vs. apoA-I within each drug condition by blocked three-way ANOVA post-hoc tests. (c) Particle size distribution of apoE-containing lipoproteins in the unconcentrated 72-h-conditioned media from drug-treated CCF-STTG1 were assessed by 6% native PAGE followed by immunoblotting for apoE. The ladder on the left represents Stokes diameter. (d-f) Cellular ABCA1 protein levels were measured by immunoblot in primary human astrocytes, HMC3 and primary human brain vascular pericytes after 72 h treatment with vehicle control DMSO, 1 μM T0901317, 3 μM AZ7235. Graphs represent fold-change over DMSO control (dashed line) and +/− 95% confidence intervals from N independent experiments indicated in brackets. *** P < 0.001 compared to vehicle control using blocked two-way ANOVA post-hoc tests. Immunoblot images were cropped to show relevant lanes