Fig. 5

Distribution of cytokeratin in stage I oocytes.

A-B) Cytokeratin (CK, green) immunostaining and DiOC6 labeling (Bb, red) in ovary tissue sections. A) In WT, CK puncta are distributed within the Bb (arrowhead) and cortically, whereas in macf1a^p6cv mutant oocyte (B-C) CKs are devoid from the cortex (white outline) and are around the nucleus. C-D) Quantification of CK enrichment in the Bb. C) DiOC6 staining pattern was used to segment the oocyte into regions of interest (ROI). Outlines show identified plasma membrane (green), nucleus (yellow) and Bb (orange). All cytoplasm defined by the area between yellow and green lines. White circles around identified CK puncta. CK puncta density was measured in each region of interest (ROI). Images are representative of 3 experiments. DAPI (blue) stains the follicle cell nuclei (A-B). Images are single optical sections. Scale bar: 20 μm. N ≥ 5 ovaries, > 30 WT and > 20 macf1a^p6cv oocytes. D) Graph of CK enrichment (CK puncta/μm²) in ROIs. N = 2 ovaries, 7 wild type and 10 macf1a^p6cv.oocytes were used for quantification. Error bars, standard error of the mean. Black bar shows the mean. P values are calculated for ROI differences by two-way ANOVA and Tukey's Multiple Comparison Text to compare between genotypes. ** P < 0.01; *** P < 0.001.