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. 2015 Mar 6;24(13):1536–1545. doi: 10.1089/scd.2014.0410

FIG. 2.

FIG. 2.

PGCs during the 3 days of in vitro AGM organ culture recapitulate the nuclear rearrangement of 5-hydroxymethylcytosine (5hmC) identified in PGCs in the embryo. (A) Immunofluorescence of E10.5 AGMs for Oct4-positive PGCs (red), 5hmC (green), and DAPI (blue). (B) AGMs cultured for 3 days to identify Oct4-positive PGCs (red), 5hmC (green), and DAPI (blue). (C) E11.5–E13.5 genital ridges stained for Mvh or Oct4-positive PGCs (red), as indicated, and 5hmC (green) and DAPI (blue). All images were obtained by using at least n=3 independent biological replicates of gonads or AGMs. (D) Quantification of 5hmC foci in individual PGC nuclei between E11.5 and E13.5. (E) Quantification of DAPI foci in individual PGC nuclei between E11.5 and E13.5. (F) Quantification of 5hmC foci that are negative for DAPI in individual PGC nuclei between E11.5 and E13.5. (G) Quantification of 5hmC foci in individual PGC nuclei during AGM organ culture on days 1–3. (H) Quantification of DAPI foci in organ culture PGCs between days 1–3. (I) Quantification of 5hmC foci that are negative for DAPI in individual PGC nuclei during organ culture on days 1–3. Statistical data were obtained by using at least n=3 independent biological replicates of gonads or AGMs. *Refers to statistical significance P<0.05. (A–C) Yellow arrow indicates 5hmC foci, white arrows indicate 5hmC territories.