Fig. 8. Early Pachytene Chromatin-associated Polyubiquitylation Patterns Are Enriched in Autosomally Synapsed Tex19.1^-/- Spermatocytes.

a Immunostaining for mono- and poly-ubiquitylated proteins (FK2 anti-ubiquitin, green) in Tex19.1^+/± and Tex19.1^-/- autosomally synapsed pachytene spermatocyte chromosome spreads. Examples of nuclei with axial and chromatin staining on the XY body are shown. SYCP3 (red) marks the lateral element of the synaptonemal complex. A single channel greyscale image for FK2 is also shown. An adjacent non-meiotic nucleus positive for FK2 is visible in the XY chromatin Tex19.1^-/- panel. Scale bar 10 μm. b Quantification of the proportions of autosomally synapsed pachytene nuclei with different sex chromosome ubiquitylation patterns. 19.1% of Tex19.1^+/± and 41.5% of Tex19.1^-/- pachytene nuclei obtained from three animals of each genotype have axial staining on the XY body (n=157, 159; χ² test, *** indicates p < 0.001). c Higher magnification and increased exposure of FK2 immunostaining showing examples of staining on autosomal axes. Examples of autosomes with clouds, foci and no FK2 staining are shown. SYCP3 (red) marks the lateral element of the synaptonemal complex. Scale bar 5 μm. d Quantification of the proportions of autosomally synapsed pachytene nuclei with clouds, foci or no FK2 staining on their autosomes. Autosomally synapsed pachytene nuclei were scored for autosomal FK2 staining regardless of the FK2 staining pattern on the sex chromosomes. 2.6% of Tex19.1^+/± and 21.4% of Tex19.1^-/- pachytene nuclei obtained from three animals of each genotype have clouds of FK2 staining on their autosomal axes, and 7.0% of Tex19.1^+/± and 23.3% of Tex19.1^-/- pachytene nuclei have foci of FK2 staining on their autosomal axes (n=157, 159; χ² test, *** indicates p < 0.001).