(A) Co-IP of FLAG-SPAG4 and LAIMINB1 with SEPT12-GFP. The cell lysates of the NT2D1 cells, transfected with pEGFP-SEPT12 and a pFLAG-SPAG4 vector, are subjected to IP with an anti-GFP antibody (right panel, Row 3) or a nonspecific control IgG (right panel, Row 2), followed by IB with an anti-GFP, anti-FLAG, or anti-LAMINB1 antibody. An input protein (5%) was used as a control during the IB of the transfected cell lysates (right panel, Row 1). (B) An IFA revealed multiple localizations of SEPT12 and LAMINB1 signals. A total of 32 male germ cells were were evaluated as (A) Round spermatids (n = 9; labeled with anti-SEPT12 and anti-LAMINB1antibodies as the presented figure,9/9); (B) elongating and (C) elongated spermatids (n = 11;11/11); (D) ejaculated spermatozoa (n = 10;10/10). (A–D) Anti-SEPT12 antibody (left panel; green), anti-LAMINB1 antibody (middle panel; red), and a combination of the left and middle panels (right panel). The arrow indicates SEPT12 or LAMINB1 signals. Scale bar: 5 μm.