viRNA size distribution varies among DENV2-infected Aag2 and C6/36 cell cultures and <i>Aedes aegypti</i> mosquitoes.

<p>Shown are Aag2 (5 dpi) library, C6/36 (5 dpi) library, <i>A. aegypti</i> (9 dpi) library. Red bars, negative-sense viRNAs; blue bars, positive-sense viRNAs. Note differences in Y-axes among graphs.</p

Northern blot hybridization to detect expression of <i>dicer2</i> mRNA in cultured mosquito cells.

<p>Total RNA from Aag2 or C6/36 cell cultures was fractionated by agarose gel electrophoresis, transferred to a nylon membrane, and hybridized to either an <i>A. albopictus dcr2</i> probe (left) or an <i>A. aegypti dcr2</i> probe (right). The biotinylated probes were detected with the BrightStar BioDetect Kit.</p

DENV2 strain Jamaica 1409 grows to higher titers in C6/36 than in Aag2 mosquito cell lines.

<p>Cell cultures were infected at a MOI of 0.001 and aliquots of medium were removed at 24 hour intervals and titrated by plaque assay. The assays were performed in triplicate and titers are expressed as means ± SEM.</p

viRNA size and genome coverage distributions vary among CFAV-infected Aag2 and C6/36 cell cultures.

<p>(A) viRNA size distribution. (B) viRNA coverage across CFAV genome for each library. Shown are Aag2 (5dpi) library, C6/36 (5dpi) library. Red bars, negative-sense viRNAs; blue bars, positive-sense viRNAs. Note difference in Y-axes among graphs.</p

DENV2 viRNAs from whole mosquito and mosquito cell culture libraries.

<p>DENV2 viRNAs from whole mosquito and mosquito cell culture libraries.</p

Logo analysis of DENV2 viRNA from mosquitoes and cell culture and CFAV viRNAs from cell culture.

<p>Logo analysis was performed on DENV2 and CFAV viRNAs using Weblogo 3. (A) Aag2 DENV2 (5 dpi) viRNA logo. (B) <i>A. aegypti</i> DENV2 (9 dpi) viRNA logo. (C) C6/36 DENV2 (5 dpi) viRNA logo. (D) Aag2 CFAV viRNA logo. (E) C6/36 CFAV viRNA logo.</p

viRNA genome coverage distribution varies among DENV2-infected Aag2 and C6/36 cell cultures and <i>Aedes aegypti</i> mosquitoes.

<p>viRNA coverage across DENV genome for each library. Shown are Aag2 (5 dpi) library, C6/36 (5 dpi) library, <i>A. aegypti</i> (9 dpi) library. Red bars, negative-sense viRNAs; blue bars, positive-sense viRNAs. Note differences in Y-axes among graphs.</p

Size and abundance of small RNA reads Mmapping to the viral genomes.

<p>The abundance of 19–30-mer sRNA reads mapping to the WNV (A), SINV (B) and LACV (C) genomes based on size. Abundance is represented as a percentage of the total viRNAs from each sample. The black bars correspond with samples collected from S2 cells and white bars from C6/36 cells.</p

viRNA coverage of the TE3′2J SINV genome in C6/36 and S2 cells.

<p>Complete genome of TE3′2J SINV (11,385 nt.) showing intensity at each nucleotide of the genome in C6/36 (A) and S2 (B) cells. Plotted are the 19–30-mer viRNA reads. Reads originating from the genomic, positive strand are represented in blue above the x-axis and those originating from the negative strand are represented in red below the x-axis. The green vertical line represents the location of the subgenomic promoter.</p

viRNA coverage of the LACV/Human/1960 strain genome in C6/36 and S2 cells.

<p>Complete genome of LACV/Human/1960 strain showing intensity at each nucleotide of the genome in C6/36 (A,C,E) and S2 (B,D,F) cells. A and B correspond with the L gene segment (6,980 nt), C and D the M gene segment (4,526 nt), and E and F to the S gene segment (984 nt). Plotted are the 19–30-mer viRNA reads across the length of each segment represented by the x-axis. Reads originating from the genomic, negative strand are represented in red below the x-axis and those originating from the positive strand are represented in blue above the x-axis.</p