Research Article

MiR-146b Mediates Endotoxin Tolerance in Human Phagocytes

Figure 3

IFNγ inhibits the upregulation of miR-146b in tolerized monocytes. (a and b) Monocytes were cultured for the indicated times with 100 ng/mL LPS (circle), 20 ng/mL IFNγ (square), or both stimuli (triangle). (a) MiR-146b and (b) miR-146a were quantified by qPCR. (c to e) Human purified monocytes pretreated or not with 10 ng/mL IFNγ, primed with 0.1 ng/mL LPS (1°LPS) for 18 h, and then challenged with 10 ng/mL LPS (2°LPS). After 24 h stimulation levels of (c) miR-146b and (d) miR-146a were assayed by qPCR in triplicate samples, normalized relative to U6, and expressed as fold enrichment as compared to unstimulated samples. (e) TNFα levels were measured by ELISA in cell-free supernatants. Results are expressed as mean ± SEM of three independent experiments.
(a)
(b)
(c)
(d)
(e)