Induction of Inflammation by West Nile virus Capsid through the Caspase-9 Apoptotic Pathway
Figure 2. Construction and subcellular expression of West Nile virus (WNV)–NY1999 capsid (Cp) gene–expressing plasmid, pcWNV-Cp-DJY: a, Genomic organization of WNV-NY1999 (10,945 bp) is outlined based on the published (GenBank accession no. AF202541). b, Cloning strategy for WNVCp gene–expressing plasmid, pcWNV-Cp-DJY. c, In vitro translated and immunoprecipitated 35S-labeled WNV-Cp visualized by SDS-PAGE. WNV-Cp–specific protein synthesis was compared to control generated by the vector backbone pcDNA3.1 (-). d, Protein expression by Western blot analysis, of WNV-Cp expression in HeLa cells. Subcellular location of WNV-Cp protein, in HeLa cells transfected with pcWNV-CpWT (e,f) or pcWNV-Cp-DJY (g,h) plasmids. 16 h posttransfection, the cells were visualized by indirect immunofluorescence. Typical nuclear staining was observed with the cells expressing WNV-CpWT (e) compared to the cells expressing WNV-Cp-DJY (g). TUNEL assay on the WNV-Cp–transfected cells, indicating nuclear condensation (i) due to expression of capsid (j).