Fig 3. AURKA is specifically required in oocytes to complete meiosis I.
(A) Representative confocal images of oocytes and eggs retrieved from oviducts of WT and Aurka KO females or oocytes matured in vitro. Cells were immunostained with antibodies against α-Tubulin (green) and DAPI (gray) (B) Quantification of percentage (%) of cells ovulated at Metaphase II (Met II); (Unpaired Students t-Test, two-tailed, **** p<0.0001). (C) Quantification of the % of oocytes that undergo polar body extrusion (PBE) in vitro (Unpaired Students t-Test, two-tailed, **** p<0.0001). (D) Quantification of the % of oocytes that undergo nuclear envelope breakdown (NEBD) in vitro (Unpaired Students t-Test, two-tailed, p = 0.6707). Graphs show the mean ± SEM from 3 independent experiments (3 females/genotype). Scale bars: 10μm. (E) Live cell light-sheet imaging of WT and KO oocytes expressing histone H2B-mCHERRY (magenta) and stained with SiR-tubulin (green). Some KO oocytes also expressed exogenous AURKA-EGFP, AURKB-EGFP or AURKC-EYFP (gray). Maximum intensity z-projections and selected time points are shown. White arrow indicates AURKC-EYFP at aMTOC. Scale bars: 10 μm. (F) Data in (E) was used to quantify % of oocytes arrested at Metaphase I. Graph shows the mean ± 95% confidence interval. n (WT, KO, KO + Aurka, KO + Aurkb, KO + Aurkc) = 10, 10, 13, 15, 11, respectively.