Figure 8. Inflammatory signaling factors TLRs, STAT/IRF as well as IFNs were verified by RT-qPCR. (A–E) RAW264.7 cells were treated for 12 h with RU.521 (2 μg/mL) or with DMSO as vehicle control. (F) RAW264.7 macrophages were pretreated with RU.521 for 12 h and then incubated for another 6 h with LPS (10 ng/mL) plus IFNγ (20 ng/mL) that polarizes macrophages to M1. Relative expression by RT-qPCR analysis. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, compared with control. Data are means ± SD. LPS, Lipopolysaccharide; IFNγ, interferon gamma; RT-qPCR, real time fluorescence quantitative PCR.