<i>In vivo</i> AeSCP-2 transcription in the larval midgut under the influence of transfected expression vectors.

<p>The siRNA vector is driven by the <i>Drosophila hsp70</i> short promoter. Constitutive over expression is drive by the Baculovirus immediate early gene promoter <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046948#pone.0046948-Vyazunova1" target="_blank">[17]</a>. In the co-transfected groups, two expression vectors were added at 1∶1 ratio. In the single expression vector groups, pBS vector plasmid was added in at 1∶1 ratio to normalized the total amount of expression vector each group received. Larvae were synchronized on Day 1 2^nd instar; siRNA expression was induced via heat shocked at 37°C for 24 hours on Day 1 of 2^nd and 4^th instar, respectively. Heat shock-treatment was not applied to the EGFP and SCP-2EGFP groups. Day 1 4^th instar larvae (10 larval midguts/sample) were taken after the 2^nd heat shock-treatment. Relative (vs. <i>rpL8</i>) AeSCP-2 mRNA levels from each sample were determined via RT-qPCR. Mean and standard deviation are shown (N = 3). The same letters above the bars in each construct represent that the mean values did not differ from other constructs significantly (p>0.05) in paired t-tests with the vector control.