Fig. 6

O’nyong nyong arbovirus infection influences expression of candidate piRNA genes in Anopheles coluzzii. Anopheles coluzzii mosquitoes were challenged with O’nyong nyong virus (ONNV) by feeding an infectious bloodmeal or an uninfected control bloodmeal, and small RNAs expressed during the primary infection at 3 d post-bloodmeal were sequenced. Analysis using Cuffdiff highlighted two candidate piRNA genes that displayed decreased abundance of mapped small RNAs in ONNV infected samples (see Results, piRNA loci XLOC_012931 and XLOC_012762). Here, the small RNA sequence reads mapping to the two candidate piRNA loci were quantified using the Integrative Genomics Viewer normalized to the library size, and the difference between ONNV infected and uninfected samples tested statistically. X-axis indicates candidate piRNA locus, y-axis indicates percentage of normalized small RNA reads mapping to the piRNA gene. ONNV-infected mosquitoes, red bar; uninfected control mosquitoes, black bar. Experiments were done in two biological replicates, error bars indicate standard deviation. Locus XLOC_012931, Chi-squared = 77.36, df = 1, p-value< 2.2e-16 (ONNV-infected mean mapped reads = 36 ± 141,421,356, mean total reads = 19,193,551 ± 8,555,908.61, ONNV-uninfected mean mapped reads = 160 ± 14,1,421,356, mean total reads = 19,167,336 ± 3,962,902.88052); and locus XLOC_012762, Chi-squared = 75.78, df = 1, p-value< 2.2e-16 (ONNV-infected mean mapped reads = 51 ± 19,09, mean total reads = 19,193,551 ± 8,555,908.61, ONNV-uninfected, mean mapped reads = 184 ± 848,528,137, mean total reads = 19,167,336 ± 3,962,902.88)