Figure 2
CRISPR-induced mutation results in molecular ablation of Adad1 and Adad2. (A) Adad1 expression in adult testes of wildtype (WT) and homozygous Adad1 mutants assessed by qRT-PCR. N ≥ 3, error bars—standard deviation. (B) Western blot detection of ADAD1 (~ 70 kDa) in adult wildtype (WT) and Adad1 mutant (M) testes. GAPDH (34 kDa) shown as a loading control. Allele of interest indicated above blot. Each lane represents an independent biological replicate. Asterisks indicate additional bands specific to the tm1Reb allele recognized by the ADAD1 antibody. (C) Adad2 expression in wildtype (WT) and Adad2em3/em3 adult testes assessed by qRT-PCR. N ≥ 3, error bars—standard deviation. (D) Western blot detection of ADAD2 (~ 70 kDa) in adult Adad2 wildtype (WT), heterozygous mutant, and homozygous mutant testes. GAPDH (34 kDa) shown as a loading control. E. ADAD2 immunofluorescence in wildtype and Adad2 mutant adult testis. Bar—100 um. * p-value < 0.01, ** p-value < 0.001, N.S.—not significant.
