Figure 5
Detection and functional analysis of TSVP-8 and TSVP-42.
(A) Immunoblot analysis of C. vestalis venom (lane 1), C. vestalis whole body without a venom apparatus (lane 2), PBS (lane 3), conditioned medium from a culture containing teratocytes (lane 4) and culture medium without teratocytes (lane 5). The upper blot was probed with a rTSVP-8 antibody, the middle blot was probed with a rTSVP-42 antibody and the lower blot was probed with a ß-actin antibody. (B) Immunoblot analysis of cell-free hemolymph from parasitized P. xylostella (P), pseudo-parasitized P. xylostella (PP) and non-parasitized P. xylostella (NP). Samples for each treatment were collected at 1, 3, 5 and 7 days and probed with either the TSVP-8 (left blots) or TSVP-42 antibodies (right blots). (C) Melanization inhibition assay. Cell-free hemolymph was collected from non-parasitized P. xylostella larvae, parasitized P. xylostella larvae, larvae injected with PBS only, or larvae injected 3× with 0.1–10 μg/ml of rTSVP-8 protein in PBS. Each bar in the graph indicates melanization level ± SE from three independent experiments. Different letters above a given bar indicates significantly different means (p < 0.05).
