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1.
FIG. 3.

FIG. 3. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

IL-8 expression in HepG2 cells requires replication competent DEN2V. UV- and heat-inactivated virus, untreated virus, and C6/36 supernatants were added to HepG2 cells for 5 days. Supernatants were analyzed for IL-8 protein by ELISA.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
2.
FIG. 4.

FIG. 4. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

NS5 and DEN2V replicon induce IL-8 expression from HEK293A cells. (A) Plasmids expressing DEN2V proteins or replicon were transfected into HEK293A cells. Supernatants were harvested day 2 posttransfection and analyzed for IL-8 protein by ELISA. (B) IL-8 mRNA levels from HEK293A cells transfected with NS5 plasmid, replicon, or untransfected cells were assessed by Taqman PCR on day 2 posttransfection. Results are normalized to untransfected cells (= 100).

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
3.
FIG. 5.

FIG. 5. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

NS5 and DEN2V replicon do not activate the ISRE or RANTES promoter. HEK293A cells were transfected with a vector containing a reporter gene for ISRE, RANTES, or IL-8 and (A) cotransfected with an expression vector for NS5 or replicon or (B) infected 24 h later with DEN2V. After 24 h (NS5 and replicon) or 72 h (DEN2V), luciferase reporter gene activity was measured and data were normalized to transfection efficiency with Renilla reniformis luciferase.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
4.
FIG. 1.

FIG. 1. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

Chemokine mRNA expression from HepG2 cell line infected with DEN2V. HepG2 cells were infected with DEN2V at a multiplicity of infection of 1. Total RNA was isolated for each time point after viral infection. Detection and quantification of the indicated human chemokine mRNAs were analyzed by RNase protection assay as outlined in Materials and Methods. P, probe lane; D, day; +, infection with DEN2V; and −, mock infection.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
5.
FIG. 6.

FIG. 6. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

Transcription factors activated by NS5 and DEN2V. HEK293A cells were cotransfected with vector encoding a reporter gene for IL-8, c/EBP, AP-1, or NF-κB and (A) an expression vector for NS5 or (B) infected after 24 h with DEN2V. After 24 h (NS5) or 96 h (DEN2V), luciferase gene activity was measured and data are normalized to transfection efficiency with R. reniformis luciferase. There is a statistically significant difference (P < 0.01) between DEN2V- and C6/36-treated cells for the IL-8, c/EBP, AP-1, and NF-κB reporters.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
6.
FIG. 2.

FIG. 2. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

IL-8 gene expression in PBMC of patients with acute dengue virus infection. Total RNA was isolated from serial PBMC samples obtained from 16 subjects with dengue fever (DF) and 10 subjects with dengue hemorrhagic fever (DHF). IL-8 and β-actin mRNA levels were measured by quantitative RT-PCR using TaqMan primers and probes. IL-8 mRNA levels are expressed as the difference in Ct between β-actin and IL-8. Values represent the mean ± standard error of the mean. F/u, follow-up visit, approximately 1 week after defervesence.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.
7.
FIG. 7.

FIG. 7. From: Dengue Virus Nonstructural Protein NS5 Induces Interleukin-8 Transcription and Secretion.

Effect of NS5 expression or DEN2V infection on transcription from luciferase-mutated IL-8 promoters. HEK293A cells were cotransfected with vector encoding a reporter gene for IL-8, mtc/EBP, mtAP-1, and mtNF-κB and (A) an expression vector for NS5 or (B) infected after 24 h with DEN2V. After 24 h (NS5) or 96 h (DEN2V), luciferase gene activity was measured and data are normalized to transfection efficiency with R. reniformis luciferase. There is a statistically significant difference (P < 0.01) between DEN2V- and C6/36-treated cells for the IL-8, mtc/EBP, mtAP-1, and mtNF-κB reporters. mtc/EBP, mtAP-1, and mtNF-κB represent the 133-luc plasmid with point mutations in the NF-IL-6, AP-1, and NF-κB binding sites, respectively.

Carey L. Medin, et al. J Virol. 2005 September;79(17):11053-11061.

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