Figure 2. The
pat-6 Gene Encodes the C. elegans Actopaxin Ortholog (A) Molecular cloning of
pat-6 (see Results for details). (B) Immunoblot of whole-worm protein extracts prepared from the wild-type N2 strain (lane 1) or a
pat-6(
st561) mutant strain that is rescued by a
pat-6::gfp transgene (lane 2). The blot was probed with anti-PAT-6/actopaxin. (C) PAT-6::GFP expressed by a rescued 430 min postfertilization
pat-6(
st561) embryo is localized to body wall muscle attachments (arrows). (D) Same embryo as in (C) immunostained with anti-PAT-6/actopaxin and MH27. As expected, muscle attachments stain with anti- PAT-6 (arrows). (E) Merge of (C) and (D), yellow regions show colocalization of the PAT-6::GFP and anti- PAT-6/actopaxin signals. (F) A nonrescued
pat-6(
st561) embryo at the same developmental stage has no GFP signal. (G) Same embryo as in (F) immunostained with anti-PAT-6/actopaxin and MH27. No PAT-6/actopaxin is detected in the region containing a dorsal muscle quadrant (arrows). Hypodermal cell junctions (arrowheads) and hypodermal cell junction remnants (labeled 'j') are stained by antibody MH27. (H) Merge of (F) and (G). Scale bar, 5 um.