Global transcriptional increase after E(z) inactivation. (A,B) Expression levels of PcG target genes and silent non-PcG target genes in EZ2-2 cells at 25°C and 31°C were assayed by RT-qPCR and shown relative to RpL32. Since the expression of RpL32 increases approximately twofold at 31°C (Supplemental Fig. 5A), this normalization is conservative. (C) Expression levels of a panel of silent intergenic non-PcG target regions in EZ2-2 cells at 25°C and 31°C. The Ras3 panel shows expression levels in a control wild-type cell line (Ras3) at the same temperatures. (D) Global derepression in coding regions after E(z) inactivation. The log ratio of the expression levels at 31°C and 25°C was calculated for non-PcG targets with different expression levels and for PcG targets. For silent genes, one expression tag read was added to the number of RNA-seq tags to estimate the approximate fold-differences. The bottom and top of each box represent the first and third quartiles, and the mark inside the box is the median value. Whiskers are extended to 1.5× interquartile range. (E) Preferential transcriptional increase at H3K27me2-enriched genes. Expression fold-change between 31°C and 25°C was calculated for the non-PcG target genes with the lowest and the highest levels of H3K27me2.
