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. 2022 Feb 22;135(4):jcs259196. doi: 10.1242/jcs.259196

Fig. 3.

Fig. 3.

Loss of ADAD2 results in altered abundance of two key DNA damage response proteins. (A) Western blot of MDC1 and BRCA1 in wild-type and Adad2M/M 21 dpp testis lysate shows reciprocal impacts on protein abundance in Adad2 mutants. GAPDH and ADAD2 blots shown as loading and genetic controls (three samples/genotype). The ADAD2 genotype control and GAPDH loading control blots are also shown for the same protein sample panel in Fig. 1D, Fig. S5B,D and Fig. S6D. (B) Immunofluorescence of BRCA1 in adult wild-type and Adad2M/M testis sections by stage (indicated by Roman numerals) showing increase of XY-body-associated BRCA1 in mutant mid- to late-stage (VII and IX) spermatocytes relative to wild type. Overlays of BRCA1 (green), SYCP3 (red) and DAPI (blue) are shown. Single channel is BRCA1 alone. Inset location is indicated by a square. (C) Immunofluorescence of MDC1 in adult wild-type and Adad2M/M testis sections by stage showing loss of MDC1 in late meiotic spermatocytes (stages IX and XII). Overlays of MDC1 (green), SYCP3 (red) and DAPI (blue) are shown. Single channel is MDC1 alone. Inset location is indicated by a square. Asterisks indicate metaphase spermatocytes, which are indicative of stage XII.