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. 2016 May 9;213(3):315–328. doi: 10.1083/jcb.201510117

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© 2016 Kern et al.

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Figure 1. — SKAP has distinct mitotic and testis-specific isoforms. (A) Western blot showing the molecular mass of endogenous SKAP from HeLa cells assessed by protein marker migration. (B, top) Immunoprecipitation mass spectrometry (IP-MS) data pooled from LAP-Astrin IPs analyzed for Astrin and SKAP peptides. (bottom) Identified SKAP peptides mapped against the SKAP amino acid sequence (ID: Q9Y448-1). (C, left) Map of the 5′ end of the SKAP locus with RNA-sequencing reads from Human BodyMap 2.0. (right) Schematic of the transcript for the short (mitotic) SKAP isoform. The underlined lysine indicates the first peptide identifiable in a tryptic digest for short SKAP. (D) Confocal image of long (testis) SKAP IF localization to elongating spermatids in a mouse seminiferous (Stage I) tubule. The seminiferous tubule lumen is indicated. (top right) Zoom-in of elongating spermatids. (E) Testis section showing IF long (testis) SKAP localization to elongating spermatids in a mouse seminiferous tubule transitioning from developmental Stage X to XI. (left) Tubule region imaged with a 40× objective. (right) Boxed region (Stage X) imaged using a 100× objective to indicate DNA localized puncta (∼6× zoom). Bars, 20 µm.