Fig. 5

Ectopic expression of loqs2 in larval stages results in larval growth arrest. A Cassettes for ectopic expression of loqs2 under the control of the Ae. aegypti polyubiquitin (PUb) promoter or the baculovirus promoter OpIE2. Fluorescent markers under control of the promoters 3xP3 or PUb were used to drive eGFP or DsRed2 expression as transgenesis markers as indicated. Images are representative of larvae from each transgenic line. B Wild-type (non-transgenic) sibling larvae, reared at the same water container, showed normal development until pupal stage while transgenic larvae exhibited developmental arrest at L2-L3 stages of development at day 5 post-hatching. C Stacked bar plots comparing development of wild-type and transgenic mosquitoes ectopically expressing loqs2 under control of the baculovirus promoter OpIE2. D–F sRNA sequencing analyses of larvae L2 ectopically expressing loqs2 compared to wild-type siblings. D sRNA abundance (18 to 35 nt) from wild-type (WT) and transgenic (Trg) larvae L2 colored by genomic origin. E sRNA abundance (18 to 26 nt) that mapped to a curated miRNA reference [76]. F sRNA abundance (18 to 35 nt) that mapped to a de novo generated siRNA cluster reference. Bar colors in E and F represent the first 5' nucleotide base distribution. sRNA abundances in D–F are reported as reads per million mapped (RPM). G Gene set enrichment analysis (GSEA) of larvae ectopically expressing loqs2 compared to wild-type siblings showed a major transcriptional shutdown on the transcription of metabolic pathways in transgenic individuals