Fig. 4

Fig. 4 Mios is required for oogenesis and nucleolar maturation.

a Schematic of the Mios protein and mutagenized regions (black arrowheads) in the Mios^ms20 and Mios^sa proteins. Truncated proteins are shown below with highlighted DNA mutations and corresponding genotyping assays. The * in Mios^sa indicates the induced stop codon. b–d Germ cell (Ddx4, teal) and DNA (DAPI, white) in mios^ms20/ms20 (50 dpf; n = 5), mios^sa/sa (45-51 dpf; n = 4), and mios^ms20/sa (50 dpf; n = 4) fish. Developing sperm are outlined with dashed lines and scale bar is 50 µM. e Sex ratio graph for 60 dpf+ wildtype, heterozygous, and mutant fish. Pink represents female fish and blue indicates male fish. Number of fish screened are indicated for each group. Statistical significance was evaluated by performing a two-tailed χ² test with Bonferroni correction against the +/+ control group. P = 0.0125. f, g Fibrillarin (yellow) immunostaining in 35 dpf wildtype (n = 7) and mios^-/- (n = 10) germ cells. Enlarged views of boxed regions show representative localization of Fibrillarin in wildtype and mios^-/- oocytes. Scale bar for all images is 20 µM. h Quantification of the Fibrillarin puncta area normalized to oocyte nuclear area for wildtype (n = 9 cells from 3 individual fish) and mios^-/- (n = 9 cells from 3 individual fish) oocytes. For box and whisker plots, line indicates median, boxes indicate upper (75th) and lower quartiles (25th), whiskers indicate data within 1.5 times the inner quartile range, and error bars represent minimum and maximum values. Any data outside of this range are plotted as individual points. Two-tailed paired equal variance Student’s t tests were performed for the indicated groups. P = 0.05. i, j p-Ubtf (active RNA pol I; purple) and RNA pol II (yellow) immunostaining in 35 dpf wildtype (n = 4) and mios^-/- (n = 4) germ cells. Enlarged views of boxed regions show representative localization of p-Ubtf (red) and RNA pol II (white) in wildtype and mios^-/- oocytes. Scale bar for all images is 20 µM. f, g, i, j Germ cells are labeled by Ddx4 (teal) and nuclei are labeled with DAPI (white). Source data are provided as a Source Data file.