Expression pattern of acsp30 in A. culicifacies female mosquito. Determination of relative transcript abundance of ascp30 in R and S strain adult mosquitoes by semi-quantitative RT-PCR (A) and real-time PCR (B). (C) Expression of acsp30 at various stages of mosquito development, second instar (2*), third instar (3*), fourth instar (4*) and pupa, is depicted as fold-induction over levels present in naïve adult female mosquitoes by real-time PCR. (D) Temporal induction of acsp30 upon Plasmodium infection. Refractory mosquitoes were fed on uninfected blood (open bars) and on P. vinckei infected blood (shaded bars) and transcript levels of acsp30 were measured by real-time PCR at different time intervals post-blood meal (PBM). For all realtime RT-PCR experiments, RNA isolated from body tissue (abdomen and thorax) of mosquitoes was used as template and expression levels of acsp30 were measured by using the Comparative CT Method. Transcript levels were normalized to the internal control, β-actin and shown as fold induction relative to the naïve adult female mosquitoes. Representative data (mean ± S.D.) from three independent experiments are shown.