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. 2020 Jan 21;27(7):2176–2190. doi: 10.1038/s41418-020-0493-4

Fig. 5. MDC1 is dispensable for NBS1’s function at meiotic prophase.

Fig. 5

a Immunofluorescence staining analyses of MDC1 and NBS1 in Mdc1+/+ and Mdc1−/− MEFs after treatments of 10 µM bleomycin for 6 h or 5 µM etoposide for 2 h. γH2AX marks DNA damage sites. Scale bar, 10 µm. b Immunofluorescence staining of SYCP3 and γH2AX were used for analyses of stages of meiotic prophase in spermatocytes from Mdc1−/− mice. Scale bar, 5 µm. Surface spreads of zygotene spermatocytes from Mdc1+/+ and Mdc1−/− mice were incubated with antibodies against RAD51 (c), DMC1 (d), RPA2 (e), and MEIOB (f). Scale bar, 5 µm. g Localization of MDC1 in surface spreads of pachytene spermatocytes from Mdc1+/+ and Mdc1−/− mice. Scale bar, 5 µm. h Localization of endogenous NBS1 in surface spreads of pachytene spermatocytes from Mdc1+/+ and Mdc1−/− mice. Scale bar, 5 µm. Cartoons depicting proteins (red) localized on chromosome loops and axes (green) are shown on the right. Arrows mark chromosome axes and arrowheads mark chromosome loops. i Localization of endogenous NBS1 in surface spreads of pachytene spermatocytes from H2ax+/+ and H2ax−/− mice. Scale bar, 5 µm. Cartoons depicting proteins (red) localized on chromosome loops and axes (green) are shown on the right. Arrows mark chromosome axes and arrowheads mark chromosome loops.