Fig 5. Progeny viruses derived from Wolbachia colonized cells replicate poorly in naïve vertebrate cells.

(A) Schematic representation of the experiment. CHIKV expressing mKate fluorescent protein from a second sub-genomic promoter was grown in C710 Aedes albopictus cells in the presence (W+ virus) or absence (W- virus) of Wolbachia (wStri strain). These progeny viruses were then used to infect naïve vertebrate BHK-21 cells synchronously at an MOI of 5 particles/cell. Virus growth in cells, plated on a ninety-six-well plate, was measured in real time by imaging and quantifying the number of red cells expressing the virus encoded mKate protein over a period of 42 hours using live-cell imaging. (B) Color of the data points distinguish the progeny viruses used to initiate infection in BHK-cells; blue represent progeny viruses derived from C710 cells without Wolbachia (W- virus) while red represent progeny viruses derived from C710 cells with Wolbachia (W+ virus). Y-axis label (Virus Positive Cells/Image) represent red cells expressing virus-encoded mKate fluorescent protein in a single field of view, four of which were averaged/sample at every two-hour time point collected over the course of infection. Two-way ANOVA with Tukey’s post-hoc test. Error bars represent standard error of mean (SEM) of biological replicates (n = 3–4).