Supplemental Figure 1. Knockdown effect of Toll and spz genes.

<strong>Supplemental Figure 1. </strong>Knockdown effect of Toll and spz genes. Effects of knockdown of the Toll (A)-(E) and spz (F) genes by RNAi were examined by qRT-PCR. On day 0, pupae were injected with 100 ng of dsRNA (malE, Toll1−4, or spz1−7) or two Toll dsRNAs (100 ng each), or a mixture of spz1−7 dsRNA (spzAll; 100 ng each), followed by incubation for 3 days. The mRNA levels of Toll or spz genes in the samples were measured by qRT-PCR. (A) Samples injected with a single Toll dsRNA. (B) Samples injected with <em>Toll1</em> dsRNA and another <em>Toll</em> gene dsRNA. (C) Samples injected with <em>Toll2</em> dsRNA and another <em>Toll</em> gene dsRNA. (D) Samples injected with <em>Toll3</em> dsRNA and another <em>Toll</em> gene dsRNA. (E) Samples injected with <em>Toll4</em> dsRNA and another <em>Toll</em> gene dsRNA. (F) Samples injected with seven spz gene dsRNAs. Experiments with three animals were independently repeated three times, and each column represents the mean ± S.D. Asterisks and double-asterisks indicate P &lt; 0.05 and P &lt; 0.01 compared to control (malE dsRNA-treated) samples, respectively, according to Student's <em>t</em>-test.