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. 2011 Sep 19;108(41):17028–17033. doi: 10.1073/pnas.1112288108

Fig. 3.

Fig. 3.

TcKnk is mislocalized after disruption of chitin synthesis. (A) Cryosections of T. castaneum pharate adult lateral body walls (20 μm thick) of control TcVer (dsVer), TcKnk (dsKnk), and TcChs-A (dsChs-A) dsRNA-treated insects were immunostained with D. melanogaster Knk antiserum (green). (B) Distribution of TcChs-A was determined by using anti–TcChs-A antibody (green) in various dsRNA-treated insects. Specificities of the D. melanogaster Knk and TcChs-A antibodies as well as the chitin probe were ascertained by using insects subjected to RNAi for these two genes as controls. The punctate distribution of both TcChs-A and TcKnk within cells suggests vesicular localization. (C) Nikkomycin (1 mM, 0.2 μL per insect) was injected either once at the pharate pupal stage or twice (on days 1 and 3 of the pupal stage) before cryosectioning and immunostaining for TcKnk (green) and staining for chitin (red). Chitin, red; proteins, green; DAPI, blue; C, cuticle; E, epithelial cells. (Scale bars: 5 μm.)