Fig. 3

GCNA Mutant Germ Cells Experience Replication Stress (A) Frequency of microsatellite repeat length changes within the C. elegans vab-1 locus. (B and C) RPA foci (yellow arrows) accumulate in Drosophila GcnaKO germ cells. (B) Control and GcnaKO/Df Germaria (left) and nurse cell nuclei (right) stained for DNA (blue), RPA (green), and Lamin C (red) as indicated. Scale bars represent 10 μm. (C) Quantification of RPA punctae per nucleus. 13-49 nuclei were counted per genotype per stage in triplicate. (D) Quantification of γH2Av foci in region 1 of the Drosophila germarium in control and GcnaKO mutant germaria in response to HU treatment. 37-52 germaria were counted per genotype per stage in triplicate. Mean values are shown with standard deviation. (E) The dvc-1/Spartan locus of C. elegans showing the strategy for creating a complete null, dvc-1(ea65). (F) Brood analysis (total viable adult offspring) and embryonic lethality associated with N2, dvc-1 and gcna-1 single mutants, and dvc-1;gcna-1 double mutant worms. n > 6 broods/genotype, 3-way Kruskal–Wallis test, p < 0.05. (G) Examples of DAPI-stained, diakinesis oocytes from dvc-1, gcna-1, and dvc-1; gcna-1 mutant germ cells show chromosome fragmentation in the double mutant. Scale bars represent 5 μm.