HSV-2 induced production of IFN-β in HCE cells is TLR4-dependent.

<p><u>(A) Knockdown of TLR4 expression.</u> The HCE cells were transfected with shRNA-TLR4 plasmid (refer to iTLR4) or scrambled shRNA control. Forty-eight hours after transfection, the total RNAs were collected and the expression of TLR4 was analyzed by RT-PCR. The expression of GAPDH was used as the internal control. The experiment was repeated three times, the representative images from one experiment are shown. <u>(B) TLR4-dependent IFN-β expression by HSV-2 infection</u>. HCE cells were transfected with shRNA-TLR4 or vector control. At 48 h after transfection, HCE cells were infected with HSV-2 at 3 MOI or mock-infected. Cell-free supernatants were collected at 0 h, 4 h, 6 h, 16 h post-infection for measurement of IFN-β secretion by ELISA analysis. Each assay was performed in triplicate. Results are reported as means ± SD. ** indicates p<0.01 and * indicates p<0.05.