Fig. S2.

Confirmation, orientation, and colocalization of CYP4G16 with CPR. (A) Expression of CYP4G16 upon RNAi-based silencing of the corresponding gene. Immunolocalization of CYP4G16 to longitudinal cryosections from mosquito specimens that were injected with dsGFP (Upper) or ds4g16 (Lower). (Scale bars, 50 μm.) (B) Whole-mount staining using α-CYP4G16 and α-CPR, in the presence or absence of Triton X-100. (Upper) Triton X-100 was used in parallel with primary antibodies, and green fluorescence was detected. (Lower) Triton X-100 was not included, and the signal was lost for both proteins (CPR and CYP4G16), indicating the absence of protein epitopes outside the cells and the intracellular orientation of the CYP4G16. (Scale bars, 50 μm.) oe, oenocytes. (C) Double staining on abdominal walls with α-CPR and α-CYP4G16. Colocalization of CPR (green) and CYP4G16 (red) appears yellow. TOPRO stains the nucleus blue.