Fig. 6

The upregulation of ABCD1 in human macrophages entering pro-inflammatory resolution is mediated by LXRα. A, B Macrophages derived from healthy control donors were treated with LPS and incubated for the indicated time. The expression of NR1H3 (encoding LXRα) and CH25H normalized to HPRT1 was analysed by RT-qPCR (n = 6). C Macrophages from 4 healthy donors were treated with either 25-hydroxycholesterol (25-HC), the LXR antagonist GSK1440233 or the solvent EtOH for 24 h followed by analysis of ABCD1 expression by RT-qPCR. D Macrophages from 5 healthy donors were treated with the LXR-agonist T0901317 or solvent control for 24 h and ABCD1 levels normalized by HPRT1 were determined RT-qPCR. E Immunoblot analysis of macrophages treated with either 25-HC, the LXR-agonist T0901317 or solvent control to determine ABCD1 protein levels normalized to β-actin (n = 4). Representative blot of macrophages derived from one healthy donor is shown. F Macrophages derived from three healthy donors were treated with either LPS or LPS and the LXR antagonist GSK1440233 for 3 or 24 h before ABCD1 mRNA levels normalized for HPRT1 were determined by RT-qPCR. One-way ANOVA and Fisher’s LSD comparison test was used for statistical analysis in A and B. Ratio paired Student’s t test was performed on absolute values in C–E and paired Student’s t test in F. ***p < 0.001; **p < 0.01; *p < 0.05; ns not significant