Fig. 4

RNA interference (RNAi)-induced suppression of Fasciola hepatica calmodulins FhCaM2 and FhCaM3 in juvenile fluke. Western blots were used to measure suppression of both FhCaM2 and 3 relative to a loading control (anti-actin) at both 7 (a) and 14 (b) days following exposure to CaM double stranded (ds)RNA. All three FhCaM targets were measured in untreated controls, negative control dsRNA treatments (dsCTRL) and a dsRNA cocktail targeting all three fluke CaMs (dsFhCaM1-3). Reduced band density, indicating suppression of target protein, is apparent in FhCaM2- and FhCaM3-probed dsFhCaM1-3 treatments at both 7 and 14 days post dsRNA, as evidenced by densitometric analyses of FhCaM2 (c) and FhCaM3 (d). Each lane represents a single replicate sample, consisting of protein extracted from 50 NEJs per replicate. *, P < 0.05; **, P < 0.01