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

Figure EV3. Generation of M1ap −/− mice.

Figure EV3

  1. The strategy to generate M1ap −/− mice. The single guide RNA (underlined) was designed to target exon 4 of the M1ap gene. A 10‐base pair deletion was detected in the M1ap knockout allele. WT, wild‐type. PAM, protospacer adjacent motif.
  2. Gel electrophoresis of the PCR products obtained from testis cDNA and subsequent Sanger sequencing showing the 10‐base pair deletion in the M1ap −/− mice. Actb served as an internal control. The arrowhead indicates the mutation site.
  3. Western blotting with testis lysates from WT and M1ap −/− mice using the anti‐M1AP antibody. GAPDH was used as the loading control. The band corresponding to full‐length WT M1AP protein is indicated by an arrow.
  4. Immunofluorescence staining of M1AP (green) and SYCP3 (red) on the spermatocyte spreads of WT and M1ap −/− mice. Scale bars, 10 μm.