FIGURE 7.

Analysis of Septin 12 interacting proteins reveals positive interaction with Protamine 2 and Lamin B2/3. (A) Co-immunoprecipitation of Protamine 2 with Septin 12 in mouse testes lysate. Blue arrows indicate a molecular weight standard; orange arrows indicate a precipitate in predicted molecular weights. Isotype IgG control shows no precipitate. (B) Co-immunoprecipitation of transfected HEK cells with mouse Septin 12 and Lamin B1, Lamin B2, or Lamin B3. NT - non-transfected cells; T - Transfected cells; Input-only cell lysate (without Co-IP); Iso - Isotype control; NC - Negative control for Co-IP (without ab); DI - Depleted input (the last wash of Co-IP). (C) 3D illustrative images showing positive transfection of HEK cells with Septin 12-GFP (green) and Lamin Bs-Myc (red). Nuclei are stained by DAPI (blue). Scale bar = 5 µm. (D) Western blot shows no significant difference in the relative abundance of Lamin B2 and Lamin B3 in mouse testes. Blue arrows indicate the molecular size of the antibody-detected bands. (E) qPCR of mouse testes shows no significant difference in relative gene expression of Lamin B2, Lamin B3, and SUN4 in testicular cells of WT and Prm2^−/− mice. n = 8, Mann-Whitney test. (F) Immunohistochemistry of mouse SUN4 (green) and Lamin B2 (red). Scale bar = 10 µm.