Figure 1

miR-181b and IL-6 are differentially expressed during LPS induced tolerance in RAW264.7 cells. (A) RAW264.7 cells were primed with 100 ng/ml LPS (1st LPS) continuously for 24 h and washed twice with PBS, cells were incubated in fresh complete culture medium for 12 h with 100 ng/ml LPS (2nd LPS) for 12 h. Levels of total (intracellular and secreted) IL-6 protein were measured by ELISA. (B). miRNAs targeting 3′-UTR of IL-6 were predicted by miRanda and RNA22. miRNAs predicted by both miRanda and RNA22 were tagged by triangle. (C) RAW264.7 cells were treated as in (A). miR-181a, miR-181b, miR-181c and let-7 g were determined by qRT-PCR. Sample data were normalized to U6 mRNA. The samples were assayed in triplicate. Data represent the mean ± S.D. from at least three independent experiments.