Figure 1.
Fetal histone H3 undergoes modification in response to a maternal high-fat diet. (a) Western blotting with antibodies specific to H3AcK9, H3AcK14, and H3AcK18 and (b) H3K4me2, K9me2, K9me3, and K27me2me3 were employed to determine the relative degree of lysine-specific acetylation (a) and methylation (b) after normalization to total histone H3. Histones were acid extracted from fetal hepatic tissue from control (n=3) and high-fat diet (n=4)-exposed animals in year 2, and resolved on SDS-PAGE gels. Relative densitometry was used in quantification (mean±S.E.M). Shown are representative bands of a minimum of three separate experiments. (c) Acid-extracted histones were resolved on an acidic urea gel under conditions of reverse polarity to enable resolution of post-translationally modified histone variants by virtue of their charge difference. Thus, retardation reflects increasing modified acetylation states following western blotting (e.g., anti-H3AcK14, left panel). Relative densitometry was used in quantification for the determination of the ratio of the arbitrarily designated ‘A’ versus ‘B’ variants AcK14A/B (mean± S.E.M). Shown are the representative bands of a minimum of three separate experiments.