Figure 5

Four mosquito-specific miRNAs that are expressed in all four species of three highly divergent genera. MiRNAs examined include miR-M1 (A), miR-1175 (B), miR-1890 (C), and miR-1891 (D). Expression was examined across the developmental stages of An. stephensi, An. gambiae, Ae. aegypti, and T. amboinensis. The top panels are northern results and the bottom panels are RNA gel images for verification of small ribosomal RNA and tRNA integrity and loading of total RNA. Emb, pooled embryos between 0-36 hr after egg deposition; L1+2, pooled 1^st and 2^nd instar larvae; L3+4, pooled 3^rd and 4^th instar larvae; Pupae, mixed pupae; F, adult females one to five days after emergence; M, adult males 1-5 days after emergence. 15 μg of total RNA per sample for An. stephensi, An. gambiae, and Ae. aegypti were used. 10 μg of T. amboinensis total RNA per sample were used. For T. amboinensis northerns, 3^rd instar larvae were not included. For the T. amboinensis miR-M1 northern, hybridization and washes were carried out at 49°C instead of 42°C to reduce background across the membrane.