The defects of hai1 mutants are phenocopied by overexpression and rescued by knockdown of matriptase1a. (A) In situ hybridization revealing matriptase1a (labelled mat1a) expression in the epidermis, olfactory epithelium (olf), otic epithelium (ot), gut (gt) and lateral line primordium (llp) at 24 hpf. (B) Counter-staining with p63 antibody (brown), demonstrating expression in the basal epidermal layer. (C,D) Lateral views of yolk sac extension of heat-shock-treated embryos injected with pTol2-hse-GTP/matriptase1a alone (C), displaying epidermal dissociation, or co-injected with pTol2-hse-GTP/matriptase1a and matriptase1a MO (D), displaying normal epidermal morphology; 24 hpf, overlays of fluorescent and Nomarski images. Cells with transgene expression are labeled by GFP. (E,F) Lateral Nomarski images of a 24 hpf un-injected wild-type embryo (E) and matriptase1a morphant (F), demonstrating that the loss of Matriptase1a does not affect epidermal morphology. (G,H) Lateral Nomarski images of hai1a mutants injected with either matriptase1a MO alone (G) or with matriptase1a and hai1b MOs (H); both display normal epidermal morphology. (I) Anti-p63 immunostaining of a 24 hpf hai1a mutant co-injected with matriptase1a and hai1b MOs. (J) Acridine orange (AO) staining alone (lower panel) and super-imposed with a Nomarski image (upper panel) of the yolk extension region of a 24 hpf hai1a mutant injected with matriptase1a MO. (K) Stills of time-lapse Movie 8 in the supplementary material (at the times indicated after 24 hpf), demonstrating that matriptase1a MO injection restores the epithelial properties of fluorescently labeled hai1a morphant basal keratinocytes. (L,M) Lateral views of the trunk and tail of an un-injected hai1a mutant (L) and of a hai1a mutant injected with matriptase1a MO (M) after in situ hybridization for the leukocyte marker lcp1 at 24 hpf.
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