Zebrafish homolog establishes and maintains cell adhesion and tissue integrity

NBP and Numbs morphants show defects in development.

Panels compareNBP morphants (NBP MO),NBP control morphants (NBP MOC) andNumbsmorphants (NumbsMO)
with wild-type embryos (WT). (A) 24 hpf embryos injected with eitherNBP MO or NumbsMO are delayed in grow th and abnormally developed in comparison with wild-type
embryos. Arrows point to dissociated cells in the dorsal region of the NBP morphants (lateral view, anterior towards the left). (B) Western blot analysis of protein extracts from
24hpf wild-typ e and NBP MO-inje cted embryos with the N1 polyclonal antibody against NBP. The wild-type extract gives a NBP-positive band of the expected size (99.57 kDa),
whereas only a negligible amount of NBP is seen in extracts from the morphants. (C, E– H) Anomalous development of the epidermis. The epidermal cells of the NBP MO-injected embryos varied in size and shape, were separated by large intercellular spaces (C) and frequently were released from the surf ace (E , asterisk denotes the gap below the
epidermal cells; arrowheads in F and H). Tumor-like structures emerged from the dorsal part of the embryo (as terisks in F, G, and H). They were composed of enlarged cells
with round,β-catenin strongly positive nuclei (F) and with α-catenin (G), β-catenin (F) and E-cadherin (H) enriched in the cell – cell contacts. Stars point to inner tissue located
just underneath the tumor-like structur es where cells lack β -catenin in both nuclei and membranes (F) and α-catenin in membranes (G). Arrows point to junctions between
neighboring epidermal cells enriched in β -catenin (F), α-catenin (G) and E-cadherin (H) in WT and to mislocalized E-cadheri n in epidermal cells of NBP MO (H). Nuclei in H
are staine d by acridine orange. Scale bar: A, D = 100μm, C, F– H= 20μ m, and E = 1 μm.

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