Table1_Identification of dihydromyricetin as a natural DNA methylation inhibitor with rejuvenating activity in human skin.pdf

Changes in DNA methylation patterning have been reported to be a key hallmark of aged human skin. The altered DNA methylation patterns are correlated with deregulated gene expression and impaired tissue functionality, leading to the well-known skin aging phenotype. Searching for small molecules, which correct the aged methylation pattern therefore represents a novel and attractive strategy for the identification of anti-aging compounds. DNMT1 maintains epigenetic information by copying methylation patterns from the parental (methylated) strand to the newly synthesized strand after DNA replication. We hypothesized that a modest inhibition of this process promotes the restoration of the ground-state epigenetic pattern, thereby inducing rejuvenating effects. In this study, we screened a library of 1800 natural substances and 640 FDA-approved drugs and identified the well-known antioxidant and anti-inflammatory molecule dihydromyricetin (DHM) as an inhibitor of the DNA methyltransferase DNMT1. DHM is the active ingredient of several plants with medicinal use and showed robust inhibition of DNMT1 in biochemical assays. We also analyzed the effect of DHM in cultivated keratinocytes by array-based methylation profiling and observed a moderate, but significant global hypomethylation effect upon treatment. To further characterize DHM-induced methylation changes, we used published DNA methylation clocks and newly established age predictors to demonstrate that the DHM-induced methylation change is associated with a reduction in the biological age of the cells. Further studies also revealed re-activation of age-dependently hypermethylated and silenced genes in vivo and a reduction in age-dependent epidermal thinning in a 3-dimensional skin model. Our findings thus establish DHM as an epigenetic inhibitor with rejuvenating effects for aged human skin.</p

Additional file 3: of The gut microbiota drives the impact of bile acids and fat source in diet on mouse metabolism

Figure S3. Impact of experimental feedings and microbial colonization on the liver. a Liver to body weight ratio and corresponding regression analysis. b Liver histopathology. c Hepatic triglyceride concentrations. For detailed description of the statistical analysis see the “Methods” section. (PNG 576 kb

Additional file 5: of The gut microbiota drives the impact of bile acids and fat source in diet on mouse metabolism

Table S1. OTU-table based on high-throughput 16S rRNA amplicon analysis. Data were obtained and analyzed as described in the text. Data are sequence counts after quality checks. Only those OTUs occurrding at > 0.25% relative abundance in at least one sample were retained. Columns are individual mice per dietary groups as abbreviated in the text and in other illustrations. (PNG 78 kb

Additional file 7: of The gut microbiota drives the impact of bile acids and fat source in diet on mouse metabolism

Table S2. List of microbial genes differentially expressed in the cecum of mice fed high-fat diets supplemented with primary bile acids and with either palm oil (PHB) or lard (LHB) as fat source. Genes included in this table are significantly (q-value < 0.05) and substantially (>2.5-fold) overexpressed in one condition as compared to the other.Gene annotation (KEGG) refers to the Kyoto Encyclopedia of Genes and Genomes database annotation. (XLSX 34 kb

Additional file 2: of The gut microbiota drives the impact of bile acids and fat source in diet on mouse metabolism

Figure S2. Impact of experimental feedings and microbial colonization on mouse metabolism. a Body weight development over time. b Regression analysis of fasting blood insulin and leptin concentrations in P- and LHB-fed SPF mice. See the “Methods” section for description of statistical analyses. c Quantification of chromogranin A-positive (ChgA+) cells in colonic tissue sections of SPF mice from the different feeding groups. Description is as Fig. 1e. ***p < 0.01, one-way ANOVA followed by the Tukey test (performed using Graph Pad Prism). (PDF 9366 kb

Additional file 4: of The gut microbiota drives the impact of bile acids and fat source in diet on mouse metabolism

Figure S4. Colonization status of SPF and GF mice. a Cecum to body weight ratio. b Viable bacterial counts were determined by anaerobic cultivation. C 16S rRNA gene-targeted PCR of cecal content DNA from GF and SPF mice. Two representative samples per dietary group are shown for each colonization status. Bands at 1.5 kbp indicate the presence of microbes. Water was used as negative template control (NTC); number of mice: between 9 and 12 per group; for detailed description of the statistical analysis see the “Methods” section. (PNG 135 kb