An in vivo assay system for RNAi genes in Tribolium. (a) A scheme of the in vivo assay system for RNAi genes. (b) Uninjected Pu11 larvae and pupae. EGFP is expressed in the wing primordia (arrow) at the larval stage, as well as in the pupal wings. EGFP is also expressed in the eye. (c) EGFP dsRNA-injected larvae and pupae. EGFP expression is completely silenced. (d, e) Larvae and pupae that were injected with dsRNA for dsRed (d) or Tc-Ubx (e) prior to the secondary EGFP RNAi. The prior injection of dsRNA for these genes does not affect the effectiveness of EGFP RNAi. Tc-Ubx RNAi pupae show hindwing to elytron transformation (e), indicating that the RNAi is working properly.