Figure 2. LncRNA CHROME is regulated by dietary and cellular cholesterol via the LXR- transcription factor.
a, qPCR analysis of CHROME variant expression in (a) livers of African green monkeys (n=5/group) before fed a high fat moderate cholesterol diet (8 weeks) compared to baseline chow diet. Box represents 25th to 75th percentiles, with line in middle indicating the median. b-c, qPCR analysis of CHROME variant expression in (b) human HepG2 cells treated with cholesterol-cyclodextrin (10 μg/mL; 72 h) compared to vehicle, and (c) human THP-1 macrophages treated with acetylated LDL (37.5 μg/mL, 24 h) compared to untreated. Data are the mean ± SEM of 5 (b) and 4 (c) independent experiments. d, Schematic diagram of predicted LXR response elements (LXRE) in the CHROME locus (top) and relative enrichment of CHROME in LXRα/β chromatin immunoprecipitates from THP-1 cells treated with LXR agonist (T0901317, 10 μM) at each of the predicted sites. Enrichment of LXR at OSBPL6 locus is shown as positive control. Data are normalized to IgG control. e, qPCR analysis of CHROME variant expression in livers of male cynomolgus monkeys treated with LXR agonist (GW3965) or vehicle for 2 days. Data are the mean ± SEM of 5 monkeys/group. f, qPCR analysis of CHROME variant expression in primary human hepatocytes and THP-1 macrophages treated with LXR agonist (10 μM T0901317) relative to vehicle control. Data are the mean ± SEM of 3–5 independent experiments. g, qPCR quantification of CHROME (all variants) and ABCA1 mRNA in THP-1 macrophages transfected with an LXRα/β-targeting or control siRNA for 24 h and then treated with LXR agonist (10 μM T0901317). Data are the mean ± SEM of 3 biological replicates from a single experiment that is representative of 3 experiments. P-values were calculated using (b, c, f, g) two-tailed student’s t-test and (a, e) two-way ANOVA. *P≤0.05, **P≤0.01, ***P≤0.001.