Semi-Quantitative RT-PCR Analysis of SmTβRII

<p>Top panel shows agarose gel separation of the PCR products of SmTβRII (middle), SmTβRI (bottom) and the constitutively transcribed control, α-tubulin (top). Lanes are numbered and the respective stages are listed at the bottom of the bar graph. The bar graph representation shows the percentage values of the optical densities in pixels of the PCR bands of SmTβRII (gray bars) and SmTβRI (white bars) compared to the corresponding band of α-tubulin control from the same stage. Values were calculated from three independent PCR amplifications (Error bars represent standard deviation values). Samples included in the assay were the hepato-pancreas regions of the intermediate host; uninfected and 30-d–infected B. glabrata snails, which represent different stages of daughter sporocysts; parasite eggs obtained from the liver of Syrian golden hamsters infected with 5,000 cercariae; cercariae shed and collected from infected snails; and stages representing different time points during schistosome development in mammalian host, obtained by perfusion of infected hamsters for the specified time (15–45-d-old worms). Adult male and female worms, separated after perfusion, were also included in the assay. Samples were subjected to total RNA extraction and cDNA synthesis, followed by PCR amplification using specific primer pairs.