Figure 1
Mice deficient in Gykl1 or Gk2 exhibit reproductive defects. (a) Immunoblotting of Gykl1 and Gk2 protein in various tissues from C57BL/6J mice. dpp: day post partum. (b) Immunostaining of Gykl1 and Gk2 in round spermatids, elongating spermatids and spermatozoa pre-stained with 150 nM MitoTracker Red CMXRos. White arrow indicates the mitochondria in elongating spermatid. Scale bar: 10 μm. (c) Gykl1 and Gk2 KO mice were generated by CRISPR/Cas9-mediated genome editing. sgRNA sequences that target the Gykl1 and Gk2 loci are indicated here. Wild-type (WT) sequences are aligned against sequences from both mutant alleles for each mutant mouse. 'n' denotes the number of nucleotides inserted or deleted in the region. (d) Whole-cell extracts were collected from the testes of wild-type and Gykl1/Gk2 KO mice, and blotted with antibodies against Gykl1/Gk2. An antibody against GAPDH was used as a loading control. (e–g) The mean pregnancy rate (e), litter size (f) and sperm count (from the epididymis) (g) for wild-type vs homozygous (−/−) and heterozygous (+/−) Gykl1 and Gk2-deficient mice were determined and compared. Error bars represent s.d. Numbers in parentheses indicate sample size (n). Significance was determined by ANOVA. *P<0.05. **P<0.01. ***P<0.001.
