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Fig. 1. ATP4 is required for normal development of the embryonic epidermis. (A and B) Morphological analysis of the embryonic epidermis at stage 32 in control morpholino oligonucleotide (CoMO) (A) and atp4aMO (B) injected specimens. Knockdown of atp4a lead to defects in ciliation of multiciliated cells (MCCs, green) and reduced numbers of small secretory cells (SSCs, red), but without obvious effects on ion secreting cells (ISCs, yellow) or outer/goblet cells (blue). (C) Knockdown of atp4a attenuated foxj1 and foxa1 expression, but not foxi1 expression in the skin epidermis. Embryos were unilaterally injected with atp4aMO at the four-cell stage and assayed for foxj1/foxa1/foxi1 expression by WMISH at stage 15. Correct targeting was confirmed by co-injection of lineage tracer. Depicted embryos are derived from the same injected batch. Numbers in the right lower corner indicate frequency of phenotype (a, anterior; d, dorsal; p, posterior; st., stage; v, ventral). |
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S2 Expression of three major transcriptional regulators in MCCs was analyzed: multicilin (mci) ( Stubbs et al., 2012), myb (Tan et al., 2013) and foxj1 ( Fig. S2C). mci was only weakly expressed at stage 10, peaked at stage 19 and was barely detectable at later stages (25 and 32) of mucociliary development. |
