Extended Data Fig. 5: Phytate digestion promotes HDAC activity in IECs.
From: Microbiota-derived metabolite promotes HDAC3 activity in the gut
a, Number of peaks identified by ChIP–sequencing with significantly increased or decreased H3K9Ac enrichment in IECs, relative to IECs harvested from GF mice. n = 2/group; Min to max plots for each comparison (4 per bar); line at median. b, ChIP-seq for H3K9Ac at HDAC3 target genes in primary IECs isolated from GF and E. coli mono-associated mice. Peaks are normalized to reads per million mapped reads. c, InsP3 (IP3) levels in 1010 colony forming units (CFU)/ml cultures of phytaseΔ E. coli (n = 4) versus wildtype E. coli (n = 4) *P = 0.0314. d, HDAC activity of mouse colonoids treated with PBS (n = 8) or phytase-digested phytate (1 mg/ml) (n = 9) for 5 h.**P = 0.0016. e, western blot detection of HDACs in mouse colonoid lysate. For gel source data, see Supplementary Fig. 1. f, HDAC activity with inositol-1,4,5,6-tetrakisphosphate (IP4) doses as indicated. n = 3/group. **P = 0.0052 (1 μM), **P = 0.0018 (100 μM). g, Relative intracellular InsP3 levels of colonoids treated with phytase-digested phytate (1 mg/ml) -/+ 40 μM carbenoxolone. n = 3/treatment. *P = 0.0189. h, CFU measured in stool collected from mice mono-associated with E. coli or phytaseΔ E. coli. n = 3/group. i, Bacterial-specific qPCR of faeces for Enterobacteriaceae, Bacteroides, and Firmicutes. n = 3/group. j, PCR of E. coli phytase gene (appA) in stool from mono-associated mice in (h, i). All graphs, except a, are mean of biological replicates ± s.e.m.; unpaired two-tailed t test. Data were independently repeated two (e, f) or three (c, d, g–j) times with similar results. *P ≤ 0.05, **P ≤ 0.01.
