Figure 2. Cellular localization of Slc15a1 in the seminiferous epithelium of adult rat testes.

(A) Immunohistochemistry was performed using frozen testis sections. Primary antibody was replaced with normal rabbit IgG as negative control. (B–D) Specific staining for Slc15a1 appears as brownish-red precipitates. Green arrowheads point to peritubular myoid cells, red arrowheads point to interstitial Leydig cells, blue asterisks indicate Slc15a1-stained nuclei of germ cells, ‘bv’ represents blood vessel. Roman numerals indicate tubule stages. Nuclei were stained with hematoxylin. Scale bars = 150 μm in (A,B), 100 μm in (C,D). (E) Specificity of the anti-Slc15a1 antibody as demonstrated by an immunoblot of testis lysate (∼50 μg protein). M, protein ladder marker. (F,G) Immunofluorescent microscopy was performed on frozen testis sections. Co-localization of Slc15a1 (red) with tight junction (TJ) marker ZO-1 (green) or adhesion junction (AJ) marker N-cadherin (green) in the adult rat testis was shown in F and G, respectively. Corresponding merged images were also shown, but no area of co-localization was observed (no orange color). Roman numerals indicate tubule stages. Nuclei were stained with DAPI (blue). Scale bars = 100 μm. Experiments were repeated three times using testes from three rats.