Figure 5.

IR8a and OR2 pathways cooperatively regulate aldehyde perception. A IR8a and obp2a are expressed abundantly on the palp. Top, RT-PCR with wildtype 5^th instar antennal and palp cDNA; Bottom, single color in situ hybridization on the 5^th instar palp cryosection. Arrowhead indicates positive cell clusters for each gene. Bar, 100µm. B Double color in situ hybridization between IR8a and obp2a on the 5^th instar palp cryosection. Left, merged staining on the palp; Right, enlarged view from left frame area, dashed line indicating potential cell clusters from one sensilla. Bar, 25µm. C POR assays toward multiple compounds with locusts treated with water or ds RNA of IR8a. N=8-9 (30 locusts each replicate) for each assay. D POR assays toward multiple compounds with locusts treated with water or ds RNA of obp2a. N=9 (30 locusts each replicate) for each assay. E POR assays toward multiple compounds with locusts treated with ds RNA mix of IR8a and obp2a. N=6-9 (30 locusts each replicate) for each assay. F POR assays toward multiple compounds with locusts treated with individual ds RNA or mix of IR8a and Orco. N=8-9 (30 locusts each replicate) for each assay. *** p<0.001, two-way ANOVA with Bonferroni post-tests. G Semi-quantitative RT-PCR to confirm RNAi silencing efficiency and specificity for ds RNA mix of IR8a and obp2a. H Similar as G, no cross-silencing between Orco and IR8a pathways with semi-quantitative RT-PCR. Actin was used to check template quality. Paraf: Paraffin oil; IAate: iso-amyl acetate; Henol: Z-3-Hexenol; Hanal: Hexanal; Henal: E-2-Hexenal; Haone: 2-Hexanone; Hpone: 2-Heptanone; DP: 2,5-Dimethylpyrazine