Figure 8

Effects of Sl SCPx dsRNA interference on the levels of the transcripts and proteins of the Sl SCPx gene, cholesterol level in the hemolymph and molting and growth of the 6^th instar larvae. (A) A total of 4 μg of dsRNA per larva was injected into the larvae after ecdysis into the 6^th instar stage. Total RNA and proteins were extracted from the midgut of the 30 treated larvae for transcript analysis by RT-PCR and protein analysis by western blotting analysis, respectively. In the RT-PCR analysis, α-actin amplification from the same RNA samples as the target gene was used as an internal control. DPI: days post injection. (B) In the western blotting analysis 30 μg of proteins was loaded per lane and were probed with anti-Sl SCPx antibody (upper panel of B) and anti-Sl SCPx-2 antibody (lower panel of B), respectively. For the Sl SCPx treatments, expression of RNA (A) and protein (B) in two independent individual samples are shown. The control was the samples injected with GFP dsRNA (A and B). (C) The hemolymph cholesterol levels in the Sl SCPx dsRNAi treated insects and the control. Three independent replicates were performed for each data point. The concentration of cholesterol in the cells or hemolymph was calculated as μg/ml of medium or hemolymph. (D) Pupation incidences in dsRNA-treated larvae. The scale bars represent 8 mm.