Fig. 8

Immuno-localization of SSEA-1, SCA-1, PCNA, Ki-67, PTEN, 5-mC and CD166. SSEA-1 (a) and SCA-1 (b) positive clearly evident and PCNA (d) expression was massively increased in DES-treated testes compared with control (c). 5-mC was expressed in spermatogonia along the basement membrane and in spermatocytes in control testes (e) but was minimally expressed (f), completely loss (h) and also some positive cells are also observed at places (g) after DES treatment suggestive of global hypomethylation. PTEN was observed germ cells in control testis (i) whereas in atrophied testis it was minimally expressed (j) but it is completely absent in bigger testis (k) suggestive from conducive environment for tumor formation. Marked increased in Ki-67 was observed after DES treatment (m, n) compared with normal age-matched vehicle-treated control (l). CD166 -a cell surface marker for stem/progenitor cells in several cancers was minimally expressed in vehicle-treated control (o) and atrophies testis (p, q), however, massively expressed in bigger testis (r-s) with tumor-like changes. Negative control with omission of primary antibody (t). Scale: 20 µm (a–d, i–k, l, o–s) and for (e–h and m, n, t) 10 µm