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. 2013 Apr 8;8(4):e60180. doi: 10.1371/journal.pone.0060180

Figure 1. Embryos with either global or local loss of Egfr signaling have defects in epithelial morphogenesis.

Figure 1

(A–F, I, J) Cuticle preparations. Black arrows, posterior spiracles; black arrowheads, puckers in cuticle; white arrowheads, dorsal hole or scab. (G, H, K, K´) Confocal micrographs of embryos during dorsal closure (DC). (A) Dorsolateral view of wild-type embryo showing intact dorsal surface. (B) Egfrf2/Egfr2C82 embryo, selected by absence of GFP balancer chromosome, showing “curled up” phenotype. (C) Egfrf2/Egfr2C82 embryo showing large dorsal hole. (D) Embryo from temperature-sensitive Egfr1F26 stock that had been allowed to develop at 18°C before shifting to restrictive temperature of 29°C at about stage 10/11. Embryo has a bowed appearance characterized by pulling in of tail (marked by posterior spiracles) and puckering of cuticle. There is a small dorsal hole in the cuticle. (E) Embryo in which EgfrDN had been expressed in the epidermis using the 69B-Gal4 driver showing dorsal scab and mild bowing. (F) Embryo in which EgfrDN had been expressed in the epidermis using the ptc-Gal4 driver showing bowed appearance and dorsal hole, similar to the embryo in panel D. (G, H) Still images from Movie S1 showing restricted expression pattern of LE-Gal4 driver, revealed using a UAS-GFP-NLS reporter. Cell outlines were revealed through expression of a Ubi-DEcadherin-GFP transgene. (G) LE-Gal4 is not expressed at beginning of DC. (H) Midway through DC, GFP-NLS is expressed in the first two rows of cells flanking the amnioserosa (AS), visualized as GFP signal in nuclei. (I) Embryo in which EgfrDN had been expressed using the LE-Gal4 driver showing mild bowing and dorsal scab. (J) Embryo in which EgfrDN had been expressed in the AS using the Gal4c381 driver showing small dorsal hole. (K) Anti-phospho-MAPK staining of a wild-type embryo showing strong immunoreactivity in the center of the amnioserosa and lateral epidermis but little staining in dorsal epidermis and cells at periphery of the AS. (K´) Same embryo as in K with phospho-MAPK in red and cell outlines revealed with anti-phosphotyrosine (PY, green). Scale bars: 50 µm (A–J)(K, K´).