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Fig. 4. sqd and hrb27C mutants have nurse cell chromosomes that fail to disperse and this phenotype is rescued by isoform-specific transgenes of Sqd and Otu. (A) Wild-type ovariole stained with Hoechst to show the polytene (arrowhead) nurse cell chromosomes that have a blob-like appearance (double arrowhead) at stage 5 and are fully dispersed (arrow) by stage 6. (B) Wild-type stage 8 egg chamber stained with Hoechst with properly dispersed nurse cell chromosomes. (C,D) sqd (C) and hrb27C (D) mutant egg chambers at stage 8 stained with Hoechst reveal that the nurse cell chromosomes fail to disperse throughout oogenesis. (E) The percentage of polytene nurse cell nuclei in stage 6 and later egg chambers from sqd¹ mutants and from sqd¹ mutants carrying transgenes expressing the specified Sqd isoforms (A, B or S) (n>400 for each genotype). (F) The percentage of polytene nurse cell nuclei in stage 6 and later egg chambers from sqd¹ mutants and from sqd¹ mutants carrying one (n=339) or two copies (n=818) of the Otu104 transgene. (G) RT-PCR analysis of otu transcripts in previtellarial egg chambers. Although the larger otu transcript is absent from hfp mutants, it is present in sqd mutants. (H) Western analysis using an Otu antibody of previtellarial egg chambers from wild type (WT), sqd¹ and hfp mutants. sqd mutants show a reduction in the amount of the 104 kDa Otu isoform. Western analysis of the samples using a Tubulin antibody reveals equal loading of the sqd and wild-type samples.

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