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Fig. 4. Ato-EGFR signaling is required for the induction of Bar expression during furrow progression. (A-C) Bar expression level in the basal undifferentiated cells in wild-type eye disc is relatively even, although it appears to have a slightly higher level at the posterior margin and immediately posterior to the furrow (B, arrows). An area marked with a rectangle in A is magnified in B. (C) Predicted two activators for Bar expression in the basal undifferentiated cells: posterior margin signal (Hh) and furrow signal (blue arrow). P, posterior margin; MF, morphogenetic furrow. (D-F) Ato is required for Bar expression. Bar expression is absent (white arrow) immediately posterior to the furrow within ato¹ LOF clone, marked by broken white line in F. A broken yellow line marks the normal anterior boundary of Bar expression right posterior to the furrow in wild type. Bar expression is rescued in the posterior part of the ato¹ LOF clone (F, yellow arrow). Apical through basal sections of confocal images are combined in order to see Bar expression in single image. (G-I) EGFR is not activated within ato¹ LOF clones. Yellow and white arrows mark the dpERK activation within the proneural clusters in the wild-type or ato¹ mutant regions, respectively. The dpERK (dual phosphorylated extracellular signal regulated kinase) staining indicates the activation of EGFR signaling pathway. (J-L) egfr^CO LOF clone shows no Bar expression immediately posterior to the furrow (white arrow in K). In K, the broken yellow line marks the normal anterior boundary of Bar expression right posterior to the furrow in wild type. White (K) and yellow (L) lines, respectively, mark the egfr LOF clone boundaries. Ectopic Ato expression (L) is observed in the posterior region of the eye disc where Bar expression is lost (lack of green in K).

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