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Fig. 7. Ptc¹⁴ does not internalize Hedgehog. (A,B) Ectopic wild type Ptc^WTGFP (A) and Ptc¹⁴GFP (B) clones in the wing imaginal disc stained with anti-Hh antibody (red). (a,b) Detail of the clones shown in A and B. Only a few vesicles of Hh are observed in the clones expressing the Ptc¹⁴ mutant protein (Ptc¹⁴GFP) (b) compared with those that express the wild-type Ptc protein (Ptc^WTGFP, arrows) (a). (C) Ptc (green) and Hh (red) expression in a wing imaginal disc containing a ptc¹⁴ clone (labeled by the absence of ß-gal staining, blue). Ptc staining inside the clone (outlined in pale blue) has the same intensity levels than in the wild-type area (arrow, green panel). (c,c') Magnification of the boxed areas in C. Ptc and Hh colocalize in spots in the wild-type area (circles) (c). Ptc does not accumulate in spots (green) in the ptc¹⁴ clone. However, Hh vesicles are still present in the ptc¹⁴ clone (red, arrowheads) (c'). (D) ptc¹⁴ clone at the AP border (shown by the absence of green, GFP) in a shi^ts1 wing disc after 3 hours at the restrictive temperature. In the absence of dynamin-dependent internalization, Hh (red) accumulates in the Ptc^WT territories at the AP border and also in the Ptc^WT cells anterior to the mutant clone (arrowhead). However, Hh protein is not accumulated in the ptc¹⁴ mutant cells (asterisk). Note that the accumulation of Hh anterior to the ptc¹⁴ clone (arrowhead) is higher than the accumulation in wild-type territory at the same distance from the AP border (arrow). This indicates that Hh diffuses further through the ptc¹⁴ mutant cells than through wild-type cells. Scale bars: 50 µm.

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