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Figure 3: Characterization of testicular VSELs by immunofluorescence: Control mice testicular smears showing presence of nuclear OCT-4A (Ai-iii), SCA-1 (Bi-iii) positive cells. SSEA-1 positive cells were localized next to the basement membrane interspersed between Sertoli cells (Ci-iii) in control mice testicular sections. In busulphan treated mice, localization of SCA-1 positive cells was next to the basement membrane interspersed between Sertoli cells (Di-iii). Note that SCA-1 positive cells were negative for Lin and CD45 (Ei-v). Note also that all the cells observed are in the size range of VSELs i.e. between 2-6um. Busulphan treated mice smears also showed presence of nuclear OCT-4A positive cells (Fi-iii) which also stained positive for phalloidin (Gi-iv). SCA-1 positive cells in busulphan treated testis also stained positive for phalloidin (Hi-iv). Busulphan treated testicular smears also showed presence of NANOG (Ii-iii) and SOX-2 (Ji-iii) positive cells, which are pluripotency markers. NANOG positive cells also stained positive for primordial germ cell marker STELLA (Ki-iv). |