Fig. 7. Effect of esd1 mutation on histone H3 acetylation and methylation in the FLC genomic region by ChIP analysis. (A) FLC genomic region analyzed by ChIP. The white box corresponds to the promoter FLC region, gray boxes to exons and the black box to the first intron. The six FLC fragments analyzed by semi-quantitative PCR are depicted and numbered. (B) PCR products after 25 cycles of esd1-2, fve-1 and esd1-2 fve-1 mutants, using as a template DNA purified from chromatin inmunoprecipitated with antibodies against acetylated H3 (AcH3). UBQ10 was amplified during 22 cycles and used as control for DNA quantification. Fold enrichment in H3 acetylation of fve-1 over esd1-2 and esd1-2 fve-1 double mutant is shown. (C) PCR products after 25 cycles of Col, esd1-3, FRI, esd1-3FRI, Ler, esd1-2, fca-1, esd1-2 fca-1, fve-1, and esd1-2 fve-1 plants, using as a template DNA purified from chromatin inmunoprecipitated with antibodies against acetylated H3 (AcH3). UBQ10 was amplified during 22 cycles and used as control for DNA quantification. Fold enrichment in H3 acetylation of mutants over wild-type ecotypes is shown. (D) PCR products as in C, but using a as template DNA purified from chromatin inmunoprecipitated with antibodies against trimethylated H3-K4 (MeH3-K4). Fold enrichment in H3-K4 methylation of mutants over wild-type ecotypes is shown.