Fig. 6

In vivo Rin silencing reduces the CHIKV infection rate in Aedes albopictus mosquitoes. a Schematic representation of the experiment. Mosquitoes were injected with 500 ng of dsRin or dsLuc two days prior to blood feeding. On day 0, a subset of mosquitoes was sacrificed to determine the level of Rin depletion. Remaining mosquitoes were orally infected with CHIKV (10⁷pfu/ml) and sacrificed six days post infection. b Total RNA was isolated and Rin silencing was determined by semi-quantitative RT-PCR on Ae. albopicutus Rin mRNA, normalized for the internal control S7. Bars represent mean Rin mRNA values normalized to dsLuc injected mosquitoes. Error bars indicate one standard error of the mean (n = 5). c Heads and bodies from the blood fed mosquitoes were separated, homogenized and the presence of CHIKV was determined by incubating the homogenate on Vero E6 cells. Bars represent the percentage of CHIKV positive mosquito bodies and heads from both dsLuc and dsRin injected mosquitoes. Asterisk indicates significant difference (P < 0.05 Fisher’s exact test). d From all the CHIKV positive mosquito heads and bodies the viral titers (TCID⁵⁰/ml) were determined. Data points represent one individual mosquito head or body. Asterisk indicates significant difference between dsLuc and dsRin injected mosquitoes (P < 0.05, Mann Whitney test) and red data points indicate mosquitoes with positive saliva