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. 2015 Jan 29;11(1):e1004954. doi: 10.1371/journal.pgen.1004954

Figure 1. Expression and localization of SCML2 during mouse spermatogenesis.

Figure 1

(A) Domain structure of mouse SCML2 protein (XP_006528733.1). MBT, malignant brain tumor repeat; DUF3588, also called Scm-like embedded domain (SLED) [11]; SAM, sterile alpha motif. (B) Western blot analysis of SCML2 in adult mouse tissues. ACTB serves as a control. (C) Western blot analysis of SCML2 on cytoplasmic extract, nuclear extract, and chromatin from postnatal day 20 testis. SYCP2, a synaptonemal complex protein, serves as a control [44]. (D) Expression and localization of SCML2 in male germ cells. Testis sections from 2-month-old mice were immunostained with SCML2 and γH2AX antibodies. Nuclei were stained with DAPI. The inset is an enlarged view of a pachytene spermatocyte with the XY body indicated. Abbreviations: Sertoli, Sertoli cells; Spg, spermatogonium; Pa, pachytene spermatocyte; RS, round spermatid; ES, elongated spermatid. Scale bar, 50 μm. (E) Localization of SCML2 to the XY body in wild type spermatocytes. Spread nuclei were immunostained with anti-SCML2 and anti-SYCP3 antibodies. SCML2 staining alone are shown in bottom panels. XY chromosomes are shown in insets. (F) Co-immunoprecipitation of SCML2 and γH2AX from wild type (Scml2 +/Y) testicular nuclear extracts. Scml2 -/Y testis was used as a negative control. (G) Lack of preferential accumulation of SCML2 in the pseudo sex body in Spo11 -/- zygotene-like spermatocytes.