MiR-146b Mediates Endotoxin Tolerance in Human Phagocytes

Figure 5

(a) THP-1 wild-type cells were stimulated for the indicated times with 1 μg/mL of LPS. MiR-146b (black circles) and miR-146a (white circles) were measured by qPCR and their levels were normalized relative to U6. (b-d) miRT-ct and miRT-146b THP-1 cells were primed or not with 100 ng/mL LPS (b), 20 ng/mL IL-10 (c), or 50 ng/mL TGFβ (d) for 18 h and then challenged with 1 ug/mL LPS for 24 h. MiR-146b (black columns) and miR-146a (white columns) were measured by qPCR and normalized to U6 levels. (e) TNFα protein levels were measured by ELISA in cell-free supernatants of pRRL-ct (white columns) or pRRL-146b (black columns) THP-1 cells primed or not with 100 ng/ml LPS and stimulated with 1 ug/mL LPS. (f) miRT-ct (white columns) and miRT-146b (black columns) THP-1 cells were primed or not with 100 ng/mL LPS, 20 ng/mL IL-10, or 50 ng/mL TGFβ for 18 h and then challenged with 1 ug/mL LPS for 24 h. TNFα protein levels were quantified by ELISA. Results are expressed as mean ± SEM of three independent experiments.

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