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. 2023 Sep 12;12:e85365. doi: 10.7554/eLife.85365

Figure 2. Ancestral high-sugar diet (HSD) exposure increased genome-wide H3K27me3 levels in the offspring.

(A) The workflow of the chromatin-immunoprecipitation followed by sequencing (ChIP-seq) assay. Embryos of normal diet (ND) and HSD-F1 flies were collected from population cages at 25°C for 30 min, and allowed to develop for 80 min to target mitotic cycle 10–12 for ChIP-seq analysis. (B) Genome browser view of H3K27me3, H3K27ac, H3K9me2, H3K9me3, and H3 density at the HOX cluster (bxd, Ubx, and Abd-A) gene regions in embryos of ND and HSD-F1 embryos. (C) Average density plots showing the signal profiles of H3K27me3, H3K27ac, and H3K9me3 at their peaks plotted across a 4 kb window (±2 kb around the start/end of signals). (D) Average density plots (top) and heatmap (bottom) showing the distribution for the changes of H3K27me3, H3K27ac, and H3K9me3 signals for regions with upregulated H3K27me3 peaks in HSD-F1 embryos, respectively. Color bar showed the Z-score value in the heatmap.

Figure 2.

Figure 2—figure supplement 1. Ancestral high-sugar diet (HSD) exposure increased H3K27me3 levels in both maternal and offspring.

Figure 2—figure supplement 1.

(A) Genome browser view of H3K27me3, H3K27ac, H3K9me2, H3K9me3, and H3 density at the HOX cluster (bxd, Ubx, and Abd-A) gene regions in embryos of two replicates normal diet (ND) and F1 flies. (B) H3K27me3 modification levels of fly cycle 10–12 embryos were analyzed by western blot (n=3 biological replicates, each containing 250 embryos). Antibodies against H3K27me3 and H3 proteins were used in the western blot. (C) H3K27me3 modification levels of fly cycle 13 (top) and cycle 14 (bottom) embryos were analyzed by immunofluorescence staining. Antibodies against H3K27me3 proteins and DAPI were used in the immunofluorescence staining. In the images H3K27me3 is in green and DAPI in white. Scale bar, 10 μm. (D–E) H3K27me3 modification levels of bodies and ovaries of indicated flies were analyzed by western blot (n=3 biological replicates, each containing 20 flies). Antibodies against H3K27me3 and H3 proteins were used in the western blot. Data are shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Figure 2—figure supplement 1—source data 1. Raw data of the western blot and immunofluorescence staining experiments shown in Figure 2—figure supplement 1.
Figure 2—figure supplement 2. Ancestral high-sugar diet (HSD) exposure enhanced Pcl expression in the offspring.

Figure 2—figure supplement 2.

(A) Schematic diagram of PCL-PRC2 complex. Polycomb repressive complex 2 (PRC2) is a chromatin-modifying enzyme that catalyzes the methylation of histone H3 at lysine 27 and contains the enhancer of zeste [E(z)], the enzymatic component of PRC2, suppressor of zeste 12 [Su(z)12], extra sex combs, and chromatin assembly factor-1 (Caf-1). Polycomb-like (PCL) interacts with PRC2 to constitute a specific form of PCL-PRC2 complex to generate high levels of H3K27me3. (B–C) mRNA expression levels of Pcl (B), E(z), Caf-1, Su(z)12, and Esc (C) of indicated flies (n=3–6 biological replicates, each containing 3 flies). (D–E) mRNA expression levels of Pcl (D) and E(z) (E) of indicated flies’ body (n=6 biological replicates, each containing 3 flies). (F–G) mRNA expression levels of Pcl (F) and E(z) (G) of indicated flies’ ovaries (E) (n=6 biological replicates, each containing 15 flies). (H) mRNA expression levels of Utx and Cbp of indicated flies (n=3–6 biological replicates, each containing 3 flies). Data were shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Figure 2—figure supplement 2—source data 1. Raw data of the qPCR experiments shown in Figure 2—figure supplement 2.