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. 2017 Jun 15;13(6):e1006803. doi: 10.1371/journal.pgen.1006803

Fig 3. Defective polarized localization of JAM-C in spermatids of Gorasp2-/- mice.

Fig 3

(A) Confocal images of GRASP55, JAM-C, peanut agglutinin (PNA) and DAPI staining of seminiferous tubule sections of adult WT mice at stage I, II-III, VIII, X and XII. Staging was performed according to Nakata. H et al [26]. Co-polarized distribution of GRASP55 and JAM-C with partial overlap is found in step 2–8 spermatids present in stage II-VIII seminiferous tubules (arrowheads). Note that JAM-C staining overlaps with PNA acrosomal staining in some spermatids. Non-polarized membrane expression of JAM-C is found in spermatogonia (asterisks). Scale bar, 20 μm. (B) Co-immunoprecipitation of GRASP55 with JAM-C from testis lysates. Western blots were probed sequentially as indicated. Arrowheads and asterisk indicate the bands corresponding respectively to JAM-C and GRASP55. (C) Left panels: Low magnification confocal images of GRASP55, JAM-C, peanut agglutinin (PNA) and DAPI staining of seminiferous tubule sections of adult Gorasp2+/+ and Gorasp2-/- mice at stage V and VIII. Due to changes in PNA staining, seminiferous tubule staging in knock-out animals was based on morphological criteria. Scale bar: 100 μm. Right panels: High magnification confocal images of GRASP55, JAM-C, and DAPI showing the loss of polarized localization of JAM-C in spermatids of Gorasp2-/- adult mice as compared to controls (arrowheads). JAM-C expression in spermatogonia is present in Gorasp2-/- mice (asterisks). Scale bar, 20 μm.