Fig. 7.
SCFAs inhibit class-1/2 HDACs and downregulate expression of class-3 HDAC SIRT1 and the histone lysine methyltransferases EZH2 and SUV39H1. (A) HDAC class 1/2 activity (fluorescence intensity) in Jurkat T-cells that were treated with fresh medium (Med), supernatant of Ec, and filtered supernatants of Pg and Fn at 1:10 dilution for 6 hours. Equal numbers (2.5×105) of cells were used for each treatment and subsequent measurement of the relative HDACs activity, using a fluorometric assay kit of Class I&II-specific HDACs activity from Sigma-Aldrich. (B and C) Western blot detection of SIRT1, EZH2, SUV39H1, acetylated histone marks H3K9-Ac, H3K27-Ac, H2B-Ac, H4K12-Ac, repressive histone tri-methylation marks H3K27me3 and H3K9me3, and activating histone tri-methylation mark H3K4me3. 2D10 cells were treated with medium (Med) or filtered Pg supernatant at 1:10 dilution for 1 hour, 3 hours, or 24 hours. The levels of β-tubulin and total histone-3 (H3) were used as loading controls and references.