Temporal and spatial distribution of IsCDA in tick gut.

(A) The highest levels of IsCDA expression are noted during early tick attachment on the host. Ticks were collected at various times of attachment to the murine hosts, and IsCDA expression was measured using quantitative RT-PCR. (B) Antisera generated against recombinant IsCDA detect a native protein in the PM. The PM structure from the tick gut was isolated and immunoblotted with IsCDA antibodies, which recognized native IsCDA with an approximate MW of 60 kDa (arrow). Migration of the protein MW marker is indicated on the left (in kDa). (C) Cellular localization of IsCDA in the tick gut tissue. Cryosections from unfixed tick gut collected at 24 h of feeding were labeled with antibodies against IsCDA and Alexa 488-labeled secondary antibodies and processed for confocal immunofluorescence microscopy. The tick tissues were labeled with a nuclear stain (DAPI). Scale bar, 10 μm.