Supplementary Figure 8: The ⁶⁴⁷AEA⁶⁴⁹ mutation of TERB1 results in telomere localization defect of SUN1.

(a) Cell lysates were prepared with testes from 3-week-old WT, heterozygous and Terb1^AEA/AEA mice and immunoprecipitated with TERB1 antibody. Lysate samples and immunoprecipitates were blotted using antibodies as indicated. The indicated numbers below the blot images represent the relative signals of Co-IPed TRF1 normalized by the signals of IPed TERB1. Band intensity of blots was quantified by ImageJ. The number for WT sample was set as 1. Uncropped blot images are shown in Supplementary Data Set 1. (b) Localization of SUN1 (red) and SYCP3 (green) on pachytene spermatocyte spreads from WT and Terb1^AEA/AEA mice. Scale bar: 10 μm. (c) Quantification of the relative intensity of SUN1 foci at the termini of synaptic chromosomes in WT and Terb1^AEA/AEA pachytene spermatocytes. Three independent experiments with 6-week-old littermates showed similar results. More than 10 spermatocyte spreads were used for quantification for each group. (a.u., arbitrary units)