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Fig. 6 | Virology Journal

Fig. 6

From: Herpes simplex virus type 2 infection triggers AP-1 transcription activity through TLR4 signaling in genital epithelial cells

Fig. 6

HSV-2 ICP0 induced AP-1 transcriptional and TLR4 promoter activity. a ICP0 could enhance TLR4-promoter transcriptional activation, cellular AP-1 activation and c-Jun phosphorylation. For AP-1 activity assay, HEC-1-A cells were co-transfected with AP-1-luc plasmid, and HSV-2 ICP0 expression or vehicle plasmid. Cells were lysed and RLUs were determined as described after 24 h. For TLR4 promoter activity assay, HEC-1-A cells were co-transfected with TLR4-promoter-luc, and HSV-2 ICP0 expression or vehicle plasmid. And the RLUs were determined 24 h post-transfection. For detecting c-Jun phosphorylation, HEC-1-A cells were transfected with vehicle or HSV-2 ICP0 expression plasmid. After 24 h, cells were harvested. c-Jun, phosphorylated c-Jun and EGFP were determined via western blot. b Overexpression of ICP0 could up-regulate TLR4 expression. HEC-1-A cells were transfected with vehicle or HSV-2 ICP0 expression plasmid. To evaluate mRNA expression level, total RNA was extracted after 24 h, and TLR4 mRNA transcription level was determined through real-time PCR. To quantify TLR4 protein level, cells were harvested 24 h post-transfection, and TLR4 expressions were examined via western blot. All experiments were performed three times, and the representative results were shown

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