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Volume 8, Number 12—December 2002

Research

Induction of Inflammation by West Nile virus Capsid through the Caspase-9 Apoptotic Pathway

Joo-Sung Yang*1, Mathura P. Ramanathan*1, Karuppiah Muthumani*, Andrew Y. Choo*, Sung-Ha Jin*, Qian-Chun Yu*, Daniel S. Hwang*, Daniel K. Choo*, Mark D. Lee*, Kesen Dang*, J. Joseph Kim†, David B. Weiner*Comments to Author , and WaixingTang
Author affiliations: *University of Pennsylvania, Philadelphia, Pennsylvania, USA; †Viral Genomix, Inc., Philadelphia, Pennsylvania, USA;

Main Article

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).

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).

Main Article

1 These authors contributed equally.

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