Fig. 6. (A) Transcription profiles and (B) normalized PCR band densities representing relative mRNA abundance of 15 selected Lospins. Total RNA extracted from salivary glands (SG), midgut (MG), ovary (OV) and carcass (CA, remnant of the tick following removal of SG, OV and MG) dissected from 5-day fed A. americanum ticks were subjected to semi-quantitative RT-PCR using gene specific primers shown in Table 1. The 16 s rRNA PCR fragment amplified from A. americanum 16 s rRNA primers (Table 1) was used as the endogenous control. PCR band densities were determined using ImageJ software (http://rsb.info.nih.gov/ij). Determined densities were normalized using the following formula: Y=V+V(H–X)/X, where Y=normalized mRNA density, V=observed Lospin PCR band density in individual tissues (MG, SG and OV), H=highest 16 s rRNA PCR band density among tested tissues (carcass in this case, CA), X=tissue (MG, SG and OV) 16 s rRNA PCR band density. (B) Normalized band densities were plotted as percent tissue distributions.