JEV-induced NF-κB targeted gene expression in human microglial cells. (A) CHME3 cells were treated for 6 hours with NF-κB inhibitor (25 μM) or PI3K inhibitor (10 μM) using DMSO as a vehicle. This was followed by JEV infection for 24 hours. Total RNA was extracted and relative IFN-β mRNA expression was measured using qPCR in inhibitor-treated cells compared to the vehicle-treated cells. GAPDH was used to normalize the data. (B) CHME3 cells were transfected with CM, M-155 or M-146a and infected with JEV 24 hours later. Total RNA was extracted 24 hours pi and levels of various NF-κB targeted genes were studied using PCR array. Hierarchical clustering represents the co-regulated genes across the groups. Relative gene expression levels were depicted according to the colour scale (green - downregulation, red - upregulation). Gene designations are listed to the left. (C) Expression of NF-κB targeted genes expression that were found to be common (n = 19) in PCR array among three different groups (CM + JEV, M-146a + JEV, and M-155 + JEV) in CHME3 cells. An attenuated expression of JEV-induced genes was observed in miR-155 overexpressing cells. (D) Expression of IL-12B, PTGS-2, CCR5 and IL-4 was studied by qPCR. The figure shows a relative abundance of transcripts in JEV-infected, mimic-transfected CHME3 cells when compared with those in cells transfected with the mimics. GAPDH was used to normalize the data. (E) Expression of MyD88 and Ikkϵ in miR-155 or miR-146a overexpressing JEV-infected CHME3 cells as seen by western blotting. (F) Expression of different mRNAs in miR-155 overexpressing CHME3 cells following infection with JEV. CM, control mimic; DMSO, Dimethyl sulfoxide; GAPDH, Glyceraldehyde 3-phosphate dehydrogenase; IFN, interferon; JEV, Japanese encephalitis virus; M-146a, mimic 146a; M-155, mimic 155.