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Fig. 6. Dl and Ser signaling in the wing pouch needs ubiquitin ligase activity. (A-H) Wg (red) in wing pouches carrying mutant clones marked by the expression of GFP (green). In all panels, ventral is downwards. Scale bar: 40 µm; 60 µm in D,G. (A,B) Clones overexpress UAS-Dl in a wild-type (A) or mib1 (B) background. Whereas ectopic Wg is induced by dorsal and (less) by ventral (arrows) clones in A, no Wg induction is observed in B. The endogenous Wg stripe is abolished (arrowhead); this is also observed in mib1 clones without Dl overexpression (data not shown). (C) mib1 clones overexpressing UAS-Dl together with UAS-neur restore Wg induction; in fact Dl activity in the ventral compartment is enhanced (arrows, compare with A). (D,E) Clones overexpress UAS-Ser in a wild-type (D) or mib1 (E) background. Ser induces Wg exclusively in the ventral compartment (D), but not when expressed in mib1 cells (E). (F) Ser regains its activity to induce Wg in a mib1 background, if it is co-expressed with UAS-neur. Ser cannot induce Wg in the dorsal compartment, irrespective of the presence of ubiquitin ligases (D,F; arrowheads), probably owing to high Fng levels. (G) Co-expression of UAS-Dl with UAS-neur{Delta}R in a wild-type background efficiently induces ectopic Wg in both compartments, indicative of increased Dl activity (compare with A). (H) This is abolished by loss of mib1. (I) Cut is imaged (red), which is a nuclear marker for the DV boundary. mib1 mutant clones are marked by the absence of GFP. UAS-Dl is overexpressed via ptc-Gal4, which drives expression just anterior of the anteroposterior boundary (white line). In the cells posteriorly adjacent to this expression domain (to the right of the white line), ectopic Cut is detected within mib1 cells if they abut anterior wild-type cells (arrow), but not if they abut mutant anterior cells (arrowhead). Inset shows an enlargement of the boxed area; the Cut-positive nuclei, which do not contain GFP appear red. The presence of these Cut-positive mib1 mutant cells suggests that mib1+ is not needed for signal reception.