Fig. 6

Influence of insulin-like peptide/IGF signaling (IIS) on ovarian development of M. vitrata. a RNA interference (RNAi) using dsRNAs specific to insulin receptor (InR), serine/threonine-protein kinase (Akt), target of rapamycin (TOR), and Forkhead box protein O (FOXO). dsRNA (1 μg) specific to each gene was injected to 5 days old pupae (pharate adult stage). Newly emerged adults were reared with 10% sugar until 5th day. Changes in mRNA levels were monitored by RT-qPCR using β-actin gene expression as reference to normalize target gene expression level. Control RNAi (dsCON) used a viral gene, CpBV302, by injecting its dsRNA at the same dose. All treatments were independently replicated three times. Different letters above standard deviation bars indicate significant difference among means at Type I error = 0.05 (LSD test). b RNAi effect of IIS components (InR, Akt, FOXO or TOR) on oocyte development. dsInR, dsAkt, dsFOXO, and dsTOR represent specific respective dsRNAs. Ovarioles were separated from 5 days old females to count the number of previtellogenic oocytes (PV), vitellogenic oocytes (VT), and chorionated (CH) oocytes. For each treatment group, 10 females were analyzed. c Expression levels of Vg and VgR in adults treated with different dsRNAs. Expression levels of these two genes were quantified by RT-qPCR in 5 days old females. All treatments were independently replicated three times. β-Actin expression was used as reference in RT-qPCR to normalize target gene expression level. Different letters above standard deviation bars indicate significant difference among means at Type I error = 0.05 (LSD test). d Photos showing ovaries from females treated with different dsRNAs. Scale bar represents 1 mm