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Fig. 5. Induction of both -H2AX-containing S-foci and L-foci on late pachytene chromosome spreads 8 hours after irradiation at 2 Gy. (A) Late pachytene cell stained for -H2AX (green) and SCP3 (red) reveals additional signals that look like S-foci and L-foci in irradiated cells compared with untreated cells (see Fig. 1Ae). ^*, XY body. Bar, 10 µm. (B) Effect of increasing doses of -rays on -H2AX foci number in pachytene cell 8 hours after irradiation. (a,a') Dose-dependent increase was found in L-foci number (a) but not in S-foci number (a'). Each column represents the mean ±s.e.m. at early (black bars) and late (gray bars) pachytene of at least three independent experiments. Lower case letters correspond to multiple comparisons of mean number of S-foci and L-foci according to the -ray doses given at a certain pachytene stage. Lower case letters are different at significantly different P-values (P<0.05). (b,b') Time-course of disappearance of -H2AX S-foci and L-foci after 1 Gy exposure to -irradiation; L-foci (b) and S-foci (b') mean number ±s.e.m. of at least three independent experiments at early (black bars) and late (gray bars) pachytene. Counting of S-foci could not be done at 15 minutes. Statistical comparisons to the control were performed at a given pachytene stage by using the Student's t-test; ^**P<0.01, ^***P<0.001. (c,c') Concomitant induction of -H2AX S-foci and L-foci after treatment with neocarzinostatin in vitro equivalent to a dose of 0.5 Gy; L-foci (c) and S-foci (c') mean number ±s.e.m. for each experimental group (n=2). Statistical comparisons to the control at a given pachytene stage were performed by the Student's t-test. ^*P<0.05. (d) MMS cell treatment does not induce -H2AX L-foci. Bars represent mean number ± s.e.m. for each experimental group (n=5) at early and late pachytene.

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