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Fig. 2. Apical localization of PVF1 and PVF3. All panels except insets represent
optical cross-sections of third instar larval wing imaginal discs. (I) A
section of a pupal wing. Apical is towards the top, and basal towards the
bottom. (A) Apical accumulation of endogenous PVF1 in a wild-type wing
imaginal disc. (B) Similarly, PVF3 is also apically concentrated. In this
panel, live discs were incubated with the PVF3 antibody prior to fixation,
thus detecting only the extracellular PVF3. (C) Upon overexpression by
MS1096-Gal4, PVF1 maintains its apical localization, whereas PVF3
accumulates within the producing cells (D). (E,F) Overexpression of PVF1
together with PVF3 reduced the portion of apically localized PVF1, which
instead accumulated within the cells. In many of the intracellular puncta,
colocalization of PVF1 and PVF3 was observed (arrows). The inset shows a
section from the basolateral region. (G) Discs expressing sGFP show
concentration on the apical side, but after secretion, significant levels were
also detected between the cells on the basolateral region. (H) Co-expression
of sGFP with PVF3 did not alter its distribution. The inset is a section
obtained from the basolateral region. Comparison with the inset in F
demonstrates that while PVF1 is trapped within the cells, sGFP is readily
detected between the cells. (I) In a wild-type bilayered pupal wing,
accumulation of PVF1 on the apical side of both layers is observed. (J) Clones
overexpressing PVF1 (marked by GFP) were generated. PVF1 localization was
monitored at a laser intensity that detects the overexpressed protein but not
the endogenous one. Apical accumulation of PVF1, which was uniform above
expressing and non-expressing cells was observed.
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