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Fig. 5. Grp protein is reduced in grp¹ mutants. (A) Developmental profile of Grp protein levels during embryogenesis. Embryo extracts from different times AED (indicated above lanes in A) were blotted for Grp as in experimental procedures. Approximately 50 embryos were loaded per lane. The yolk proteins, visualized by Ponceau staining, confirm equal loading for the initial part of embryogenesis, but become depleted as embryogenesis progresses. (B) The grp¹ mutants have severely reduced levels of Grp protein. 6-7-hour-old grp¹ homozygous or heterozygous mutant embryos from heterozygous parents were genotyped by the absence or the presence of GFP encoded by the balancer chromosome Cyo-GFP (see experimental procedures). Extracts were prepared and blotted for Grp and for GFP (to confirm genotype). Each lane contained extracts from 25 embryos. Both wild type and grp¹/Cyo-GFP heterozygotes have comparable Grp levels, while in grp¹ mutants, Grp levels are reduced. (C) Grp protein persists in mutants homozygous for a deficiency that removes grp. 7.5-8.5-hour-old Df/Df homozygous or heterozygous mutant embryos from heterozygous parents were genotyped by the absence or presence of GFP encoded by the balancer chromosome Cyo-GFP. Extracts were prepared and blotted for Grp and for GFP. Each lane contained extracts from 10 embryos. The difference in GFP signal between B and C is due to different antibody dilutions of anti-GFP. (D) Extracts of embryos from grp¹ homozygous mothers or wild type (Wt) mothers were analyzed by western blotting. Extracts from approximately 50-75 embryos were loaded in each lane. Maternal genotypes and embryo ages are shown above each lane.

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