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. 2009 Oct 19;284(50):34590–34599. doi: 10.1074/jbc.M109.056317

FIGURE 5.

FIGURE 5.

Transfected miR-146a alone could mimic LPS priming in LPS tolerance assay. THP-1 cells transfected with 40 nm miR-146a mimic followed by incubation for 24 h, mock transfected cells, and 10 ng/ml LPS primed cells (tolerance positive control) were washed with complete growth medium and challenged with 1000 ng/ml LPS for 3 h. A, RNA isolates were prepared from cell pellets of mock and transfected cells followed by qRT-PCR for miR-146a expression normalized with RNU44. B, ELISA analysis of TNF-α protein in supernatants tolerance-positive control, mock transfected, and miR-146a mimic transfected cells. C and D, multiplex analysis of IL-1β and IL-6 proteins in supernatants of tolerance-positive control, mock transfected, and miR-146a mimic-transfected cells. E, the same RNA samples were also analyzed by qRT-PCR for mRNA expression of miR-146a targets TRAF6, IRAK-1, and an unrelated gene lamin A/C (LMNA, negative control). Data are representative of three independent experiments and expressed as mean ± S.D. *, p < 0.01 compared with mock transfected cell.