Journal: Journal of Virology

Article Title: Identification of Natural Molecular Determinants of Ross River Virus Type I Interferon Modulation

doi: 10.1128/JVI.01788-19

Figure Lengend Snippet: Multistep growth curves of RRV-T48 and RRV 2548. The growth kinetics of RRV-T48 or RRV 2548 were analyzed in Vero (A), HeLa (B), and L929 cells (C). Cells were infected with either RRV-T48 or RRV 2548 at an MOI of 0.1. All the cell culture media were collected at 12 h, 24 h, 36 h, and 48 h p.i. and analyzed by plaque assay. The limit of detection was 30 PFU/ml. Values are expressed as the mean titers ± SEMs from three independent experiments. *, P < 0.05; **, P < 0.01 using two-way ANOVA with Bonferroni’s post hoc test.

Article Snippet: HeLa (ATCC CCL-2) and mouse fibroblast L929 cells (ATCC CCL-1) were cultivated using Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% FBS.

Techniques: Infection, Cell Culture, Plaque Assay

Journal: Journal of Virology

Article Title: Identification of Natural Molecular Determinants of Ross River Virus Type I Interferon Modulation

doi: 10.1128/JVI.01788-19

Figure Lengend Snippet: RRV 2548 type I IFN induction and sensitivity in vitro. L929 cells infected with either RRV-T48 or RRV 2548 at an MOI of 0.1 or transfected with poly(I·C) were sampled at 12 h and 24 h p.i. (A) Type I IFN levels in cell culture medium were determined by IFN bioassay. (B) Type I IFN sensitivities of RRV-T48 and 2548 were assessed by growth inhibition and are presented as described for Fig. 2 except that human IFN-β was used at 0.05, 0.1, or 0.25 ng/ml. Error bars represent ± SEMs from three independent experiments. *, P < 0.05; ***, P < 0.001 using two-way ANOVA with Bonferroni’s post hoc test (A); **, P < 0.01; ***, P < 0.001 using Student’s unpaired t test (B).

Article Snippet: HeLa (ATCC CCL-2) and mouse fibroblast L929 cells (ATCC CCL-1) were cultivated using Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% FBS.

Techniques: In Vitro, Infection, Transfection, Cell Culture, Bioassay, Inhibition

Journal: Journal of Virology

Article Title: Identification of Natural Molecular Determinants of Ross River Virus Type I Interferon Modulation

doi: 10.1128/JVI.01788-19

Figure Lengend Snippet: Type I IFN modulation of chimeric RRVs. (A) The chimeric RRV M12 and M34 were constructed as shown. (B) Vero and L929 cells were infected with RRV-T48, RRV 2548, RRV M12, and RRV M34 at an MOI of 2, and cell culture media were collected at 12 and 24 h p.i. for analysis by plaque assay. The type I IFN induction (C) and sensitivity (D) of RRV-T48, RRV 2548, RRV M12, and RRV M34 were determined as described for Fig. 4A and ​andB,B, respectively. Error bars represent ±SEMs from three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 using two-way ANOVA with Bonferroni’s post hoc test (B and C); *, P < 0.05; ****, P < 0.0001 using one-way ANOVA with Bonferroni’s post hoc test (D).

Article Snippet: HeLa (ATCC CCL-2) and mouse fibroblast L929 cells (ATCC CCL-1) were cultivated using Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% FBS.

Techniques: Construct, Infection, Cell Culture, Plaque Assay

Journal: Journal of Virology

Article Title: Identification of Natural Molecular Determinants of Ross River Virus Type I Interferon Modulation

doi: 10.1128/JVI.01788-19

Figure Lengend Snippet: Levels of type I IFN signaling proteins in cells infected with RRV strains, recombinants, and mutants. L929 cells were infected at an MOI of 2 with RRV-T48, RRV 2548, RRV M12, RRV M34, and RRV-T48 harboring the changes indicated above the panels. Cell were collected at 24 h p.i., lysed, and analyzed by Western blotting using antibodies recognizing RIG-I, MDA5, IPS-1, TRAF3, TBK1, IKK-i, IRF3, p-IRF3, IRF7, STAT1, STAT2, p-STAT1, and p-STAT2. Antibody against β-actin was used to detect loading control. Blots represent one of two independent reproducible experiments.

Article Snippet: HeLa (ATCC CCL-2) and mouse fibroblast L929 cells (ATCC CCL-1) were cultivated using Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 5% FBS.

Techniques: Infection, Western Blot, Control