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. 2018 Apr 24;51(2):93–100. doi: 10.1267/ahc.17039

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2018 The Japan Society of Histochemistry and Cytochemistry

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 4. — Simultaneous localization of non-methylated and methylated GATCG sequences by HELMET in a paraffin-embedded section of mouse ovary. The paraffin-embedded section was blocked with a dideoxynucleotide mixture by TdT and then the Sau3A I cutting sites were labeled with biotin-16-dUTP. After dideoxynucleotide blockade, Mbo I cutting sites were labeled with digoxigenin-11-dUTP and then both haptens were visualized with FITC-anti-biotin (A, D and G) and rhodamine-anti-digoxigenin (B, E and H), respectively. Merged images are given in C, F and I. Red arrow pointed the nuclei of oocytes and white arrow pointed granulosa cells. And non-methylated GATCG was localized in the nuclei of granulosa cells (white arrows) and its staining became to be more intense from primary to secondary follicles. Bar = 20 μm.