Figure 4. Expression Patterns of MNR-1 and SAX-7 (A) Schematic of the
mnr-1::GFP reporter construct, which should be governed by most, if not all, regulatory elements of the
mnr-1 locus (Tursun et al., 2009). (B) Image of an adult animal, showing expression of the
mnr-1::GFP reporter in the lateral and ventral hypodermal ridges, but not in seam cells. At earlier larval stages, some expression is also seen in a limited number of neurons and the anterior pharyngeal bulb (Figures S6A and S6B). dhr, dorsal hypodermal ridge; lhr, lateral hypodermal ridge; sc, seam cells; vhr, ventral hypodermal ridge; an asterisk indicates the vulva. (C-E) Images of an
mnr-1(
dz175) mutant animal transgenically rescued by the hypodermically driven mCherry::MNR-1 fusion. The fusion is localized throughout the hypodermis in a punctate pattern (D and E). Similar punctate patterns are seen when mCherry::MNR-1 is expressed in muscle or when a SAX-7::mCherry fusion or mCherry alone is expressed in the hypodermis (data not shown). An arrow points to coelomocytes and an arrowhead points to the PVD axon. Anterior is to the right and ventral is down. Scale bar indicates 50 mm. (F) Magnification of coelomocytes, showing the vesicular red fluorescent staining of the mCherry::MNR-1 fusion (arrow). Scale bar indicates 10 mm. (G) Optical section of a nonpermeabilized HEK293 cell transiently transfected with mCherry::MNR-1. The red channel shows fluorescence of the mCherry::MNR- 1 construct, and the green channel shows localization of extracellular mCherry::MNR-1 as determined by an a-mCherry monoclonal antibody. Partial overlap likely indicates that a part of the fusion is localized intracellularly to the secretory pathway. Similar results were obtained with COS-7 cells (data not shown). DIC, differential interference contrast. Scale bar indicates 20 um. (H and I) Ventral images of adult animals of the genotypes indicated, stained with a monoclonal a-SAX-7 antibody. Arrowheads indicate sublateral lines stained with the a-SAX-7 antibody. Scale bar indicates 50 um. See also Figure S6.