High glucose inhibits induction of <i>Abca1</i> in BMDMs.

<p>Bone marrow cells from C57BL/6 mice were differentiated into macrophages under high (25 mM D-glucose) or normal (5.5 mM D-glucose + 19.5 mM L-glucose) glucose. Prior to treatments, macrophages were cultured in 1% FBS overnight and then treated for four hours with 5 μM T + 1 μM 9cisRA (T+9) or DMSO vehicle control and steady state RNA (A) and nascent RNA (B) transcripts of <i>Abca1</i> were profiled using qRT-PCR. <i>Cyclophilin A</i> was used for normalization. (C) BMDMs were cultured and treated as in A for eight hours. Whole cell protein extracts were collected and separated on a 7.5% polyacrylamide gel and were blotted for ABCA1. HSP90 was used as a loading control. (D) Whole cell protein extracts collected from untreated BMDMs cultured under high and normal glucose were separated as in (C) and blotted for LXRα. HSP90 was used as a loading control. The levels of LXRα and HSP90 protein were quantitated by densitometry and presented below with normal glucose set to 1. The ratio of panels A and B represent an average of three independent experiments. Error bars indicate the SEM. Significance is determined using the two-tailed Student's t-test (*, P < 0.05, **, P<0.01, ***, P<0.001). (C) and (D) are representative.