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. 2013 Jan 2;8(1):e53270. doi: 10.1371/journal.pone.0053270

Figure 6. DNA methylation levels during mammary gland development and differentiation at casein gene promoters.

Figure 6

(A, B) DNA methylation levels (%) for Beta-casein promoter (Csn2) (average of 4 CpG sites in 326 bp), (C,D) Alpha s2b casein promoter (Csn1s2b) (average of 2CpG sites in 300 bp) and (E,F) Kappa casein promoter (Csn3) (average of 5 CpG sites in 588 bp) were determined by bisulfite cloning and sequencing. Mammary epithelial Cell enriched fractions (MEC): Virgin (8_15V; 8–15 week old virgins,); Pregnancy (7P: 7day pregnant females); Inv (>28 day after lactation); AMV: Age Matched Virgin (Virgin animals same age as >28 day involuted animals). Whole tissue: pregnancy (16P: 16 days pregnant); Lactation (8L; 8 day lactation). Non-mammary gland: none-MEC cell fraction of the mammary gland (8_15; 8–15 Week old virgins); Pregnancy (7P: 7day pregnant females); Inv (>28 day after lactation); AMV: Age Matched Virgin (Virgin animals same age as >28 day involuted animals)); or non-mammary tissue: Liver. (A,C,E) Overview of Csn2 (A), Csn1s2b (C) and Csn3 (E) promoter and CpG sites analyzed, Tabular representation of DNA methylation levels (%) at individual CpG sites in analyzed region, and DNA methylation levels (%) heatmap representation (yellow 0%-dark blue 100%);. (B, D, F) Methylation status at CpG sites in Bisulfite-sequence clones for mammary gland or non-mammary gland tissue (yellow 0% methylation, dark blue 100% methylation): (2–6 individual preps (mammary gland MEC (8-15wV 8–15, 7dP, INV, AMV) or Non-Mammary gland epithelial cells (8-15wV_s, 8-15_s, 7dP_s, INV_s, AMV_s)) and 4–6 individual animals (16dp, 8dL, Lvr), 5–10 clones/DNA prep), Csn2 (C), Csn1s2b (E), Csn3 (F).