Journal: Biochemistry and Biophysics Reports

Article Title: Normal spermatogenesis and fertility in Spmip8 deficiency male mice

doi: 10.1016/j.bbrep.2025.102406

Figure Lengend Snippet: Generation and validation of Spmip8 knockout mice. (A) Schematic diagram of Spmip8 −/− mouse creation. (B) Sanger sequencing of genomic DNA shows a deletion in the Spmip8 - gene. (C) Spmip8 −/− mice were identified by genomic PCR. (D) Spmip8 - transcripts were not detected in adult Spmip8 −/− testes, n = 3 for each genotype. (E) Western blot analysis the SPMIP8 protein in Spmip8 knockout mice. α-TUBULIN was used as a loading control. (F) Immunofluorescence staining of SPMIP8 (green), PNA (acrosome, red) in testis sections from 10-week-old WT and Spmip8 −/− mice. Magnification ×40 in the panels. DAPI (blue) stains the nuclei. The head signal in elongating spermatids is non-specific, as it appears in both WT and Spmip8 −/− testis sections. Scale bar: 50 μm ∗∗∗ P < 0.001.

Article Snippet: The membrane was blocked with 5 % nonfat milk in TBST for 1 h, then incubated overnight at 4 °C with primary antibodies against SPMIP8 (1:1000, HPA062092, Sigma, Germany) and α-TUBULIN (1:5000, 11224-1-AP, Proteintech, China).

Techniques: Biomarker Discovery, Knock-Out, Sequencing, Western Blot, Control, Immunofluorescence, Staining