Fig. 5. DOC1R localizes to microtubules during meiotic maturation. (A-F) Localization of the endogenous DOC1R protein. Groups of 20 oocytes at different stages of meiotic maturation were fixed with formaldehyde and further stained with the anti-DOC1R antibody (AF) or with the antibody blocked with 30 µM of immunogenic peptide (G). (H,I) Oocytes microinjected with DOC1R-GFP encoding RNA and collected in GV (H) or MII (I). All oocytes were analyzed by confocal microscopy using identical settings. (A) Immature oocyte in GV, (B) oocyte collected 1 hour after GVBD, (C) 5 hours after GVBD, (D) 8 hours after GVBD, (E,G-I) 14 hours after GVBD in metaphase II (MII), (F) cultured for 13 hours after GVBD then treated with nocodazole for 1 hour (MII+NZ). The DOC1R staining or DOC1R-GFP localization appear in green and chromosomes in red. Two-hundred and fifty oocytes were scored for this experiment. Scale bar: 10 µm.