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Fig. 5. PlexB genetically interacts with MICAL and forms a complex with
PlexA. (A,B) Filleted preparations of late stage 16 embryos stained with
the anti-Fasciclin II monoclonal antibody to reveal motor axons of the ISNb.
(A) ISNb axons of plexBKG00878 heterozygous embryos
defasciculate properly and resemble wild-type embryos. (B) ISNb axons
in embryos double heterozygous for both plexB and MICAL
(Df(3R)swp2MICAL/+; plexBKG00878/+) often fail
to defasciculate properly and do not innervate their proper muscle targets
(open arrows). (C) A chart outlining the results from yeast two-hybrid
interaction assays with cytoplasmic regions of PlexA and PlexB. Numbers
indicate the amino acids of PlexA or PlexB that comprise each fragment. A
positive interaction is indicated by a plus sign (+). The C2 region of the
PlexA cytoplasmic domain (animo acids 1702-1945) and the C1 region of the
PlexB cytoplasmic domain (amino acids 1402-1784) support growth and reporter
gene transcription when grown on selective plates, whereas all other
combinations between PlexA and PlexB do not. (D) Lysates from embryos
expressing Myc-PlexB, with or without HA-PlexA, were immunoprecipitated using
anti-HA antibodies and blotted with anti-HA or anti-Myc antibodies to detect
the presence of HA-PlexA or Myc-PlexB, respectively. HA-PlexA
immunoprecipitates from embryo lysates also contain Myc-PlexB. Scale bar in A:
10 µm for A-B.