The effect of CLSP2 on transcriptional activation of <i>PPO</i> genes.

<p><b>A</b>) The effect of CLSP2 on the expression of 10 <i>PPO</i> genes in the mosquito carcass. The mRNA levels of <i>PPOs</i> were determined using real-time RT-PCR. Data were represented as mean ± SEM. *, p < 0.05; **, p < 0.01. The experiments were repeated four times. iLucBb, iLuc mosquitoes injected with <i>B</i>. <i>bassiana</i>; iCLSP2Bb, CLSP2 dsRNA-treated mosquitoes injected with <i>B</i>. <i>bassiana</i>; iCLSP2, mosquitoes injected with CLSP2 dsRNA; iLuc, luciferase RNAi-treated control mosquitoes. <b>B</b>) Immunoblot analysis of hemolymph PPO cleavage. Hemolymph was isolated at 24 h post-infection from four groups (iLuc, iLucBb, iCLSP2, and iCLSP2Bb, see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004931#sec009" target="_blank">Material and Methods</a>) of mosquitoes, separated by SDS-PAGE, and then probed with <i>Ae</i>. <i>aegypti</i> PPO3 polyclonal antibodies. A ~20-kDa band appeared in samples of iLucBb, iCLSP2 and iCLSP2Bb. <i>Ae</i>. <i>aegypti</i> Lipophorin II was used as the loading control. iLucBb, iLuc mosquitoes infected with <i>B</i>. <i>bassiana</i>; iCLSP2Bb, CLSP2 dsRNA-treated mosquitoes infected with <i>B</i>. <i>bassiana</i>; iCLSP2, mosquitoes treated with CLSP2 dsRNA; iLuc, luciferase dsRNA treated control mosquitoes.