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Fig. 4. (A) Mitotic index of primary spermatogonia (I SPG) in the frog R. esculenta testis at various times, 4°C and 8 h:16 h photoperiod. The I SPG was expressed as the number of metaphases per total primary spermatogonia counted x100 in three randomly chosen sections/animal. Values are expressed as means ± S.D. Differences were considered significant at P<0.01. (B) Western blot detection of PCNA protein in cytosolic testicular extracts of R. esculenta at various times, 4°C and 8 h:16 h photoperiod. Proteins (40 µg/lane) were resolved by SDS-PAGE, transferred to nitrocellulose membrane and then incubated with antibody raised against PCNA protein; a specific band at 36 kDa (by comparison with comigrating size markers). The blot is representative of three separate assays. (C) The amount of PCNA was quantitated by using ImageQuant 5.2 Program. The values shown are representative of three separate experiments.





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