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. 2011 Dec 22;287(8):5954–5968. doi: 10.1074/jbc.M111.289322

FIGURE 4.

FIGURE 4.

C3 transcription and secretion and surface expression of CD14 and TLR4 are differently regulated by oxLDL and LXR agonist TO191317 in human resting and classically (M1) and alternatively (M2) activated PBM-derived macrophages. A and B, the level of C3 mRNA (A) determined by real-time RT-PCR and C3 secretion (B) determined by ELISA in resting or classically or alternatively activated PBM-derived macrophages 0–48 h after the addition of oxLDL (50 μg/ml). C, level of C3 mRNA determined by real-time RT-PCR in resting or classically or alternatively activated PBM-derived macrophages 0–48 h after the addition of LXR agonist TO191317 (TO; 2.5 μm). D, the level of C3 mRNA in human PBM-derived macrophages; real-time RT-PCR; 100% is the level in the resting macrophages treated with BSA. E, ELISA of C3 protein content in the culture medium of PBM-derived macrophages. Values are presented as means ± S.E. (error bars) of six independent experiments. The statistical analyses of differences were performed separately for values in RM, M1, or M2 groups using an unpaired Student's t test (*, p < 0.05) or Dunnett's criterion (for comparison of RM, M1, and M2) (#, p < 0.05). F, level of CD14 mRNA in human PBM-derived macrophages; real-time RT-PCR; 100% is the level in the resting macrophages treated with BSA. G, FACS analysis of surface CD14 expression in human PBM-derived macrophages; the percentage of CD14+ cells is indicated. H, level of TLR4 mRNA in human PBM-derived macrophages; real-time RT-PCR; 100% is the level in the resting macrophages. I, FACS analysis of surface TLR4 expression in human PBM-derived macrophages. Medians of surface TLR4 expression are indicated. Values are presented as means ± S.E. of six independent experiments. The statistical analyses of differences between compared groups were performed using Dunnett's criterion (#, p < 0.05). PBM-derived macrophages were treated with BSA (20 ng/ml) (RM), with IFNγ (20 ng/ml) and bacterial LPS (100 ng/ml) (M1) or IL-4 (20 ng/ml) (M2) for 6 days. Cells were treated with oxLDL (50 μg/ml) and/or LXR agonist TO191317 (2.5 μm) and/or LPS (100 ng/ml) for 0–48 h.