Fig. 3. Quantification of RNA interference in A. gambiae Sua 4.0 cells and A. stephensi larvae. (A) A. gambiae Sua 4.0 cells co-transfected with pMinEGFP and one of the dsRNA-expressing plasmids. (B) Wild-type A. stephensi larvae injected with pMinEGFP and one of the dsRNA-expressing plasmids. The intensity of EGFP and DsRed expression was calculated using the Lucia G image-processing and analysis software. Red and green fluorescence levels were normalised to control experiments in which pMinEGFP and pMiRED were delivered. Black bars indicate normalized EGFP expression levels; grey bars indicate normalized DsRed expression. The plotted data show mean values ± S.E.M. from >20 fields captured in three separate transfection experiments or >20 injected larvae per experiment. All reductions in EGFP expression were found to be highly significant (P<0.01). DsRed expression levels did not vary significantly between individual experiments (P>0.05). S 465, pIR-EGFP₄₆₅S; NS 465, pIR-EGFP₄₆₅NS; Linker 465, pIR-EGFP₄₆₅-Linker; S720, pIR-EGFP₇₂₀S. (C) EGFP-silencing after injection of either pMiRED (broken line) or pIR-EGFP₄₆₅-Linker (solid line) into stable EGFP-expressing transgenic line V_B. The intensity of EGFP expression was calculated 3, 4 and 5 days post-hatching.