Fig. 4: BRME1 is a stabilizer of MEILB2-BRCA2. | Nature Communications

Fig. 4: BRME1 is a stabilizer of MEILB2-BRCA2.

From: The BRCA2-MEILB2-BRME1 complex governs meiotic recombination and impairs the mitotic BRCA2-RAD51 function in cancer cells

Fig. 4

a Schematic of the Brme1 knockout allele. Rectangle; exon. The two sgRNA-targeting sites and the location of the primers used for genotyping are shown. The gel shows the PCR results for WT and Brme1−⁄− alleles. b Immunostaining of zygotene spermatocytes. Scale bar: 5 μm. c Testes from WT and Brme1−⁄− males. Scale bar: 5 mm. Testis d or epididymis e sections from 8-week-old WT and Brme1−⁄− males stained with hematoxylin and eosin. Spermatogonia (SG), spermatocyte (SC), and spermatid (SP). Scale bar: 100 μm. f Testis sections with stage V–VI seminiferous tubules from 8-week-old WT and Brme1−⁄− males stained with TUNEL and DAPI. The percentages of TUNEL-positive seminiferous tubules (those containing more than three TUNEL-positive cells) were quantified. n shows the analyzed seminiferous tubule number pooled from two mice for each genotype. Scale bar: 15 μm. g Immunostaining of spermatocytes. Scale bar: 5 μm. h Immunostaining of Brme1−⁄− zygotene spermatocyte. Graph: the quantification of MEILB2 foci intensities in zygotene spermatocytes, normalized to the average value of WT. Red bars: mean value with SD. n shows the analyzed foci number pooled from seven cells from two mice for each genotype. Scale bars: 5 μm or 1 μm in the magnified panel. i The number of MEILB2 foci associated with the chromosome axes. Red bars: mean value. n shows the analyzed spermatocyte number pooled from three mice for each genotype. j Immunostaining of zygotene spermatocytes expressing GFP-BRCA2-MBD (a.a. 2117–2339). Scale bars: 5 μm or 1 μm in the magnified panel. k, l Immunostaining of zygotene spermatocytes. DMC1 k or RAD51 l were stained. The images in the other stages are shown in Supplementary Fig. 4c, d. Graph: the number of foci associated with the chromosome axes. Red bars: mean value. n shows the analyzed spermatocyte number pooled from three mice for each genotype. Scale bar: 5 μm. m Schematic of meiotic BRCA2 complex assembly pathways. All analyses used two-tailed t tests. ns: not significant. ****p < 0.0001. Leptotene (L), zygotene (Z), early-pachytene (EP), late-pachytene (LP), diplotene (D). Source data are provided as a Source Data file.

Back to article page