Figure 3
Accumulation of acetylated histones in chromatin of milk fat globule-epidermal growth factor 8 (MFG-E8) gene in NCI-H716 cells. (a) Schematic representation of MFG-E8 promoter from chromatin immunoprecipitation (ChIP) analysis. Three sets of PCR primers, designated as MP1, MP2, and MP3, were used for amplification of the distal and proximal MFG-E8 promoters. Nucleotide positions are numbered relative to the major transcription start site (+1) defined in the published DNA sequence. (b) Quantitative ChIP analysis of the occupancy of AcH3K9 after stimulation with 10 mM butyrate acid in different regions of the MFG-E8 promoter. The results are expressed as the percentage of immunoprecipitate over total input DNA relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Values are shown as the mean and standard deviation of results from more than three independent experiments. *P<0.05, **P<0.01 vs control (non-stimulated cells). (c) Time-dependent mRNA expression of MFG-E8 in NCI-H716 cells after stimulation with 10 μM trichostatin A, which also functions as a histone deacetylases (HDACs) inhibitor. All signals were standardized to the intensity of GAPDH. **P<0.01 vs control (non-stimulated cells).
