Skip to main content
. 2023 Sep 12;12:e85365. doi: 10.7554/eLife.85365

Figure 3. H3K27me3 modifications were essential for the transgenerational regulation of sweet sensitivity and feeding behavior upon ancestral high-sugar diet (HSD) exposure.

(A) Schematic diagram of the PRC2 germline RNAi experiment. Note that UAS-gene X RNAi was integrated into both two third chromosomes and Mat-tub-GAL4 was integrated into both two second chromosomes. The embryos of Mat-tub-GAL4>UAS gene X RNAi flies were collected and fed with normal diet (ND) (black, referred to ND controls) or HSD (red, referred to HSD-F0) until maturity. HSD-F0 flies were mated to produce the next generation (HSD-F1). These flies were prepared for biochemical and behavioral assays. (B) H3K27me3 modification levels in fly ovaries were analyzed by western blot (n=3 biological replicates, each containing 15 flies). Antibodies against H3K27me3 and H3 proteins were used in the western blot. (C) H3K27me3 modification levels in fly germarium were analyzed by immunofluorescence staining. Antibodies against H3K27me3 proteins and DAPI were used in the immunofluorescence staining. In the images H3K27me3 was shown in green and DAPI in white. Examples of oocyte nucleus were indicated by white arrowheads. Scale bar, 10 μm. (D–E) Fractions of flies of the indicated genotypes showing proboscis extension reflex (PER) responses to sucrose (n=3–5 biological replicates, each containing 8–12 flies). The S50 indicated the sucrose concentration that elicited PER responses in 50% of the tested flies. Data were shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Figure 3—source data 1. Raw data of the western blot, immunofluorescence staining, and behavioral experiments shown in Figure 3.

Figure 3.

Figure 3—figure supplement 1. H3K27me3 in the early embryo was required for transgenerational modulation of sweet sensitivity and feeding behavior.

Figure 3—figure supplement 1.

(A) Schematic diagram of PRC2 embryo RNAi experiments. The embryos of nosNGT-GAL4 flies were collected and fed with normal diet (ND) until maturity and mated to UAS-gene X RNAi flies produce the next generation nosNGT-GAL4>UAS gene X RNAi. The next generation fed with ND until maturity is called ND (black, referred to ND controls) and fed with HSD until maturity is called F0 (red referred to HSD-F0). The HSD exposure nosNGT-GAL4 flies (fed with HSD from embryo) mated to UAS-gene X RNAi flies produce the next generation and fed with ND until maturity is called F1 (pink, referred to HSD-F1). These flies were prepared for behavioral assays. (B) Fractions of flies of the indicated genotypes showing proboscis extension reflex (PER) responses to different concentrations of sucrose (n=3–6 biological replicates, each containing 8–12 flies). The S50 indicated the sucrose concentration that induced PER responses in 50% of the tested flies. (C) Fractions of flies of the indicated genotypes showing PER responses to 20% sucrose (n=10–12, each containing 5 flies). (D) Fractions of flies of the indicated genotypes showing PER responses to different concentrations of sucrose (n=3–6 biological replicates, each containing 8–12 flies). The S50 indicated the sucrose concentration that induced PER responses in 50% of the tested flies. Data were shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Figure 3—figure supplement 1—source data 1. Raw data of the behavioral experiments shown in Figure 3—figure supplement 1.
Figure 3—figure supplement 2. H3K27ac and H3K9me3 in the early embryo were not required for transgenerational modulation of sweet sensitivity and feeding behavior.

Figure 3—figure supplement 2.

(A–B) Fractions of flies of the indicated genotypes showing proboscis extension reflex (PER) responses to different concentrations of sucrose (n=3–6 biological replicates, each containing 8–12 flies). The S50 indicated the sucrose concentration that induced PER responses in 50% of the tested flies. Data were shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Figure 3—figure supplement 2—source data 1. Raw data of the behavioral experiments shown in Figure 3—figure supplement 2.
Figure 3—figure supplement 3. H3K27me3 in the germline was not required for sweet sensitivity and feeding behavior induced by high-sugar diet (HSD) exposure.

Figure 3—figure supplement 3.

(A–B) Fractions of flies of the indicated genotypes showing proboscis extension reflex (PER) responses to different concentrations of sucrose (n=3–6 biological replicates, each containing 8–12 flies). The S50 indicated the sucrose concentration that induced PER responses in 50% of the tested flies. Data were shown as means ± SEM. ns p>0.05; *p<0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
Figure 3—figure supplement 3—source data 1. Raw data of the behavioral experiments shown in Figure 3—figure supplement 3.