Figure 5From: HSI2/VAL1 PHD-like domain promotes H3K27 trimethylation to repress the expression of seed maturation genes and complex transgenes in Arabidopsis seedlingsChromatin immunoprecipitation (ChIP) and quantitative PCR (qPCR) analyses of H3K4me3, H3K9me2, H3K27me3 and H3K36me3 levels on endogenous GSTF8 promoter and transgene chromatin in WTLUCand hsi2-4LUCmutant. A. Genomic structures of endogenous GSTF8 gene and GSTF8::LUC transgene showing the locations of amplified regions by ChIP-qPCR. B. qPCR analyses of chromatin samples from 5-day old seedlings of WTLUC and hsi2-4LUC that were immunoprecipitated using either specific antibodies recognizing indicated histone methylation marks or IgG (non-specific binding control). Data is expressed as percentage of immunoprecipitated DNA relative to input DNA. ACT2/7 (H3K4me3 and H3K36me3), FUS3 (H3K27me3) and TA2 (H3K9me2) serve as positive controls whereas TA2 (H3K4me3 and H3K36me3) and ACT2/7 (H3K9me2 and H3K27me3) were used as negative controls. Data represent means (’sD) from two biological replicates with three qPCR replicates each. Significant differences in enrichment between WTLUC and hsi2-4LUC for each genomic region tested were determined using two-tailed Student’s t-test assuming unequal variances and P values are indicated by letters (a = p < 0.05, b = p < 0.005, c = p < 0.0005, d = p < 0.0001).Back to article page