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. 2015 May 21;5:10484. doi: 10.1038/srep10484

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Copyright © 2015, Macmillan Publishers Limited

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(A) TcCP30 cross-linking with TcCPR27 and TcCPR18. Proteins extracted from elytra dissected from five dsTcCPR27 (dsCPR27)-, dsTcCPR18 (dsCPR18)-, dsTcCP30 (dsCP30)- or dsTcCPR4 (dsCPR4)-treated insects (200 ng per insect) were collected at the day 0 adult stage and analyzed by 15% SDS-PAGE, followed by Coomassie staining (left panel) or western blotting (right panel). Arrows in right panel indicate large TcCP30 immunoreactive proteins with apparent molecular masses of ~37 and ~45 kDa. dsRNA for TcVer (dsVer) (200 ng per insect) was injected as a negative control. (B) Lac2 is involved in TcCP30 cross-linking. dsRNAs for TcCP30 (dsCP30) (200 ng per insect) or TcLac2 (dsLac2) (20 ng per insect) were injected into late stage larvae and 0-1 d-old pupae, respectively. Proteins were extracted from pronotum and elytra dissected from five dsRNA-treated insects collected at the 0 d-old adult stage (A0) and analyzed by 15% SDS-PAGE, followed by Coomassie staining (left panel) or western blotting (right panel). Arrows in left panel indicate TcCP30-like proteins (note that they are absent in the dsCP30 lanes). dsRNA for TcVer (dsVer) (20 or 200 ng per insect) was injected as a negative control.