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. 2022 Nov 28;24(2):e55778. doi: 10.15252/embr.202255778

Figure EV1. Generation of M1ap KI/KI mice.

Figure EV1

  1. The strategy to generate mice that carry an M1ap knockin (KI) mutation equivalent to that of our patients. The single guide RNA (sgRNA, underlined) was designed to target the splicing mutation site (written in red). In the KI allele, thymine was replaced by cytosine. Nucleotides from exon 7 are written in blue. WT, wild‐type. PAM, protospacer adjacent motif.
  2. Reverse transcription‐PCR analysis of WT and homozygous M1ap KI/KI mouse testes with primers spanning exons 6–8. Actb served as an internal control.
  3. Western blotting with testis lysates from WT and homozygous M1ap KI/KI mice using the anti‐M1AP antibody. GAPDH was used as the loading control. The band corresponding to WT full‐length M1AP protein is indicated by an arrow.
  4. Immunofluorescence staining of M1AP (green) and SYCP3 (red) on the spermatocyte spreads of WT and M1ap KI/KI mice. Scale bars, 10 μm.