Blockade of miRNA-146a expression abrogated LPS-induced tolerance in THP-1 cells. THP-1 cells transfected with 40 nm miR-146a inhibitor for 24 h, along with mock transfected, were washed with complete growth medium and primed with 10 ng/ml LPS for 16 h. Cells were then washed with complete growth medium and challenged with 1000 ng/ml LPS for 3 h. Additional controls included were cells transfected with miR-146a inhibitor alone and cells treated with 1000 ng/ml LPS alone for 3 h (LPS alone, TNF-α-positive control). A, total RNA from cell pellets of mock and miR-146a inhibitor-transfected cells, primed with 10 ng/ml LPS for 16 h, were analyzed by qRT-PCR for miR-146a expression. B, TNF-α protein in supernatants of TNF-α (+) control, miR-146a inhibitor alone (without LPS), mock transfected, and miR-146a inhibitor-transfected, were measured by ELISA. C, total RNA were analyzed for TRAF6, IRAK-1, and an unrelated gene lamin A/C (LMNA) expression in both mock and miR-146a inhibitor-transfected cells by qRT-PCR. Data are expressed as mean ± S.D. of three independent experiments. *, p < 0.01 compared with mock transfected cells.