Figure 3.

Treatment with sCD74/MIF significantly upregulates gene expression of type I interferon (IFN)‐induced genes. WT were treated with 40 nmol/L of sCD74 in the absence or presence of 8 nmol/L of rMIF. After 8 hours, mRNA was isolated. A, Microarray analysis was performed, and only genes that were at least 1.5‐fold differentially regulated upon sCD74/rMIF treatment compared with control were depicted. Independent triplicates were performed. Corresponding P values are depicted in Table 1. The 9 genes that were also significantly regulated upon MIF‐treatment were marked as overlap. Type I IFN‐induced genes and genes involved in NF‐κB signaling pathways are indicated as black and dark gray bars, respectively. Genes labeled as gray bars seem not to contribute to specialized function and pathways. B and D, RT‐qPCR was performed with the cDNA and Taqman probes specific for the type I IFN‐induced genes, interferon‐induced protein 44 (Ifi44), immunoresponsive gene 1 (Irg1), and C‐type lectin domain family 4, member e (Clec4e). GAPDH was used as a housekeeping gene. Relative quantity (RQ) values were calculated according to the ΔΔCt method and normalized to control. Data represent mean±SEM of at least 4 independent experiments and were analyzed with a 2‐tailed, unpaired t test with multiple correction (n=5). ^$ P<0.05 vs control; *P<0.05 vs rMIF. GAPDH indicates glyceraldehyde 3‐phosphate dehydrogenase; IFN, interferon; MIF, macrophage migration inhibitory factor; NF‐κB, nuclear factor kappa B; rMIF, recombinant MIF; sCD74, soluble CD74; WT, wild type.