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Figure 3 | Journal of Inflammation

Figure 3

From: MicroRNA-181b regulates endotoxin tolerance by targeting IL-6 in macrophage RAW264.7 cells

Figure 3

Up-regulation of miR-181b in LPS tolerance is NF-kB-dependent. (A) RAW264.7 cells were primed with 100 ng/ml LPS continuously for 12 h and 24 h. pri-miR-181b were determined by qRT-PCR. Sample data were normalized to β-actin mRNA. (B, C) qRT-PCR and western blots of p65 in RAW264.7 cells after p65 siRNA or negative control siRNA transfection. β-actin was used as an internal control. (D) RAW264.7 cells were transfected with p65 siRNA or negative control siRNA, 24 h later, cells were primed with 100 ng/ml LPS continuously for 12 h and 24 h. pri-miR-181b were determined by qRT-PCR. Sample data were normalized to β-actin mRNA. (E) One set of primers was used in the qPCR analysis to span the potential promoter region of miR-181b. (F) RAW264.7 cells were primed with 100 ng/ml LPS continuously for 24 h. qPCR analysis of p65 binding in the promoter region of miR-181b by ChIP assay. NC: negative control. The data were subjected to Student’s t-test. *p < 0.05, **p < 0.01.

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