Supplementary Material for: <b><i>Anopheles stephensi</i></b> Dual Oxidase Silencing Activates the Thioester-Containing Protein 1 Pathway to Suppress <b><i>Plasmodium</i></b> Development

We characterized the dual oxidase (Duox) gene in the major Indian malaria vector <i>Anopheles stephensi</i>, which regulates the generation of reactive oxygen species. The AsDuox gene encodes for a 1,475-amino-acid transmembrane protein that contains an N-terminal noncytoplasmic heme peroxidase domain, a calcium-binding domain, seven transmembrane domains, and a C-terminal cytoplasmic NADPH domain. Phylogenetic analyses revealed that <i>A. stephensi</i> Duox protein is highly conserved and shares 97–100% amino acid identity with other anopheline Duoxes. AsDuox is expressed in all the developmental stages of <i>A. stephensi</i> and the pupal stages revealed relatively higher expressions. The Duox gene is induced in <i>Plasmodium</i>-infected mosquito midguts, and RNA interference-mediated silencing of this gene suppressed parasite development through activation of the thioester-containing protein 1 pathway. We propose that this highly conserved anopheline Duox, being a <i>Plasmodium</i> agonist, is an excellent target to control malaria parasite development inside the insect host.