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Fig. 3. PtdIns(4,5)P₂ accumulation at the CP. Unfertilized dechorionated eggs were injected with mRNA encoding either PH::GFP or a mutated form PH::GFP^** whose PH domain does not recognise PtdIns(4,5)P₂. Eggs were given sufficient time to produce translated protein products (5 hours). (A) Brightfield and fluorescence images are displayed following fertilization showing the accumulation of microvilli at the CP and the enrichment of PH::GFP at the same region of the fertilized egg. Temperature 19°C, n=7 replicates, 3 animals. (B) PH::GFP^** did not localize to the PM before or during fertilization and had a more cytoplasmic distribution. This is seen in the three images showing an unfertilized egg (I), an egg 3 minutes after fertilization (II) and 4 minutes after fertilization, when the CP has formed (III). Bar, 50 µm. Temperature 19°C, n=5 replicates, 3 animals. (C) Line-scan analysis of eggs injected with PH::GFP or the mutant PH::GFP^**. PH::GFP is clearly accumulated at the CP following fertilization, while some PH::GFP also remains accumulated at the PM. PH::GFP^**, however, is not accumulated at the plasma membrane and displays a cytoplasmic distribution.

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