Fig 3

Fig 3

(A/B) Data resulting from test crosses between rad21l1^-/- males and wild-type females to assess fertility and reproductive phenotype. rad21l1^+/+ male tank mates were used as controls. No significant difference in the number of eggs the males caused the females to release, the composition of the resulting clutch at 6 hpf, or the survival of the embryos through 48 hpf. Data pooled from 14 crosses over 5 weeks using the same pool of 14 rad21l1^-/- males, 12/14 of which crossed successfully at least once. Unpaired, two-tailed student t-test used for statistical analysis, ns = p>0.05. (C) Whole mount adult testes stained for DNA (gray) and Ddx4 (green) showing a phenotypic range of gonad morphology in rad21l1^-/- males. All samples except #5 displayed large clusters of mature sperm. Images marked as A and B were taken from the same sample to show variation within a single gonad. Wild-type tank mates used as controls. Scale bar = 50 μm. (D) Testes sections stained with a PNA telomere probe (Tel; magenta), an antibody to γH2AX (green), and DAPI (blue), showing that telomere clustering and DSB localization (γH2AX) are normal in the rad21l1 mutant. Scale bar = 5 μm. (E) Fold change of relative mRNA expression for rad21l1 paralogs, rec8b, rec8a, and rad21a in wild-type versus the rad21l1^-/- mutant (Mut). Fold change was determined using the 2^-ΔΔCq method. eF1a (housekeeping) mRNA levels were used as a reference.