Temporal and spatial expression patterns of Scal\Orco (A) and Hirr\Orco (B). Temporal expression was evaluated using template synthesized separately from newly laid embryos (t1–3h or t3–4h), pre-hatching embryos (t22–24h or t11–12h), 3rd instar larvae (ll), pre-tanning/’early’ puparia (ep) and late pharate adults (lp; puparia 2 days prior to adult eclosion). Spatial expression was analyzed from female or male pooled antennae-maxillary palp (a-mp; stable fly) or antennae (ant) and maxillary palps (mp) isolated separately (horn fly), proboscis (prb), thorax (thx), abdomen (abd), and legs. Expression of biting fly ribosomal protein S3 (RpS3) was used as a positive control for presence of template.