Fig. 1.

Microarray analysis of miRNAs induced during the macrophage antiviral response. (A) WT murine macrophages were stimulated with medium (m), 2 μg/ml poly(I:C) [p(I:C)], or 1,000 units/ml IFN-β for 6 h. RNA was extracted and used to conduct a microarray analysis to determine the expression levels of 200 mammalian miRNAs. Data are presented on a scatter plot showing log₁₀-transformed signal intensities for each probe on both channels for the Cy3-labeled media controls and samples stimulated with Cy5-labeled IFN-β (Left) or poly(I:C) (Right). (B) RNA used in A was analyzed by qPCR to assay expression of miR-155 and in a separate experiment by Northern blot analysis under the same conditions. (C) RNA used in A was assayed by qPCR to detect expression of the small nuclear RNA U6 as a loading control or IP10 mRNA to ensure equivalent stimulation by poly(I:C) and IFN-β. (D) A portion of the macrophages generated in A were stimulated with medium, poly(I:C), or IFN-β and assayed for CD11b and CD86 expression by using FACS to ensure proper macrophage development and activation, respectively.