Fig 5. Immunofluorescence detection of CEH-18-expressing cells. CEH-18-expressing cells were detected using anti-CEH-18Amg57 antibodies to stain wild type (a,c,g,i), or ceh-18
) (e). Nuclei were also visualized with DAPI (b,d,f,h,j). The same samples are shown in a and b (dissected gonad of adult hermaphrodite), c and d(wild-type threefold-stage embryo(, e and f (ceh-18
) three fold stage embryo), g and h (wild-type 12-cell embryo), i and j (wild-type -150-cell embryo). There are five pairs of gonadal sheath cells per gonad arm, designated 1 - 5 (see Fig. 3). Only one of each pair is shown; the other five nuclei are out of the plane of focus but also stain. The DAPI image in b i s in a slightly lower focal plane to show the diakinesis stage nuclei in the proximal arm of the gonad (indicated by arrows). The arrows mark CEH- 18 -expressing gonadal sheath cell nuclei, which are also numbered. The distal arm of the gonad containing pachytene stage and mitotic germ cell nuclei is indicated by an arrow. Sperm nuclei are indicated. The background in the germ-line cytoplasm is attributable to the secondary antibody. No staining was seen in ceh-18
) at any stage of development (see text). Note CEH-18 is first expressed late in embryogenesis at the threefold stage, before hatching, and is absent from early embryos. At hatching, CEH-18 is most abundant in the pharyngeal muscle nuclei, the nuclei of the hypodermal syncytia (hyp 1-11), the nuclei of the seam cells, the P hypodermal blast cells, and the body muscle nuclei. CEH-18 is notably absent from nearly all neurons at this stage, although it is expressed by the anterior touch cells ALML/R. Bars, 50 and 25 um, in a and c, respectively. a and b were kindly provided by R. Francis and T. Schedl (Washington University School of Medicine, St. Louis, MO).