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Fig. 4. The absence of Pio causes cuticle detachment and tracheal defects. (A) In wild-type embryos at the end of embryogenesis, viewed by differential interference contrast, the dorsal trunk of the trachea appears smooth and rather straight. (B) Homozygous pio mutant embryos complete development but the tracheae appear twisted and broken (arrows). (C) In live wild-type embryos, the epidermis (marked with tubulin-GFP) closely follows the outline of the cuticle (viewed by differential interference contrast). (D) In homozygous pio mutant embryos, the cuticle detaches from the GFP-marked epidermis (arrow).