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Fig. 8. Localization of SPE-38 in spermatids and spermatozoa. SPE-38 was detected
using a peptide polyclonal antibody against a putative extracellular loop
domain of SPE-38 (SPE-38101–114). (A) DIC (top) and
epifluorescence (bottom) images of in vivo-activated wild-type spermatids and
spermatozoa. Permeabilized cells were cryomethanol fixed and permeabilized
with Triton X-100. Live cells were incubated with antibody prior to fixation
and were not permeabilized. Insets are enlargements of cells from the same or
equivalent fields. (B) Sperm prepared as described in A, but incubated with
anti-SPE-38 antibody that was pre-incubated with competing peptide. (C) DIC
(top) and epifluorescence (bottom) images of in vitro-activated
spe-38(eb44) and wild-type sperm. Cells were paraformaldehyde fixed
and premeabilized with Triton X-100. (D) Western blot of wild-type and
spe-38(eb44) adult males. Each lane contains exactly 400 male worms.
Equivalent loading was confirmed by Coomassie Blue staining. The sequence
coding the antigenic peptide (SPE-38101–114) is deleted in
eb44 mutants.
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