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Fig. 7. Removal of Dlp by PI-PLC treatment or by RNAi does not affect Wg
accumulation on Rab5SN-expressing cells. (A-D) Imaginal discs after 5
hours 30 minutes Rab5SN expression in the dorsal compartment (dorsal
Rab5SN-expressing tissue indicated by brackets). Images are projections of
1-µm-spaced confocal sections over 20 µm. (A) Disc mock-incubated
for 1 hour, stained for Dlp. (B) Disc mock-incubated for 1 hour,
stained for Wg. (C) Disc incubated for 1 hour with PI-PLC, stained for
Dlp. (D) Disc incubated for 1 hour with PI-PLC, stained for Wg. Wg is
not released with Dlp. (E,F) Discs subjected to Dlp RNAi for 40
hours in the dorsal compartment (indicated by brackets) stained for Dlp (E) or
extracellular Wg (F). Dlp protein is undetectable and extracellular Wg fails
to spread in dorsal cells. (G,H) Apical extracellular Wg
staining of discs in which Rab5SN has been expressed for 5 hours in the dorsal
compartment (indicated by brackets), either in the presence (G) or absence (H)
of Dlp protein. (H') shows depletion of Dlp by RNAi in the dorsal
compartment of a disc also expressing Rab5SN. (I-L) Disc ubiquitously
expressing YFP-Rab7, overexpressing Dlp in the dorsal compartment (indicated
by bracket) and stained for Wg. A projection of three sections 3 µm below
the apical membrane is shown. (I) Wg imaged with high gain, emphasizing
plasma membrane recruitment. (J) Wg imaged with low gain, emphasizing
punctate endosomal staining. (K) YFPRab7-labeled endosomes. (L)
Overlay of Wg and YFPRab7.
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