(A) Graph indicating functional group distribution of genes regulated by cactus and caspar silencing. For both upper (Cactus) and lower (Caspar) diagrams, colored sections of graphs correspond to the number of genes either up-regulated (right of zero) or down-regulated (left of zero) for the given functional group (see legend). Unk, unknown; Div, diverse; R/T/T, replication/transcription/translation; Met, metabolism; Trp, transport; C/S, cytoskeletal/structure; Rdx/St, redox/stress; Prot, proteolysis and digestion; CSR, chemosensory; Imm, immunity. Functional groups were predicted using Gene Ontology terms . (B) Venn diagram indicating comparative gene regulation between the two treatment groups. The blue and yellow circles represent the gene expression profiles of the cactus- and caspar-silenced mosquitoes, respectively, with the overlapping region (green) representing genes that were regulated by both treatments. Numbers with upward-pointing arrows indicate the number of induced genes, numbers with downward-pointing arrows indicate the number of repressed genes, and the single number with one downward-facing and one upward-facing arrow represents the one gene that was enriched after cactus silencing and repressed after caspar silencing. (C) Temporal expression analysis of anti-Plasmodium gene after Caspar depletion. Bars reflect fold change in expression of the indicated gene at 6, 12, 24, and 48 hours after injection of dsRNA against caspar. Fold change is determined by real-time quantitative PCR comparison to GFP dsRNA–treated controls. Horizontal gray line represents baseline expression (i.e., a 1∶1 expression ratio between control and silenced samples), and error bars indicate standard deviation among three biological replicates. P-values were ≤.05 for: FBN9 at 12 and 48 h.p.i.; TEP1 at 48 h.p.i.; LRRD9 at 6, 24, and 48 h.p.i. Other time points are indicative of trends but did not reach statistical significance among three replicates.