Fig. 6.
Ash1 is required for normal H3K36me2 levels at HOX and other target genes. (A) H3K36me2 bulk levels are unchanged in ash1 or MRG15 null mutants. Western blot on serial dilutions (4:2:1) of total extracts from imaginal discs from wild-type (wt), ash122 m+ z− and MRG15Δm– z– mutant larvae, probed with antibodies against H3K36me2 and, as loading control, histone H4 and Caf1. The genotype of the MRG15Δm− z− animals is MRG15Δ/Df(3R)BSC741 (see text). (B) ChIP qPCR analysis in wild-type (dark-green bars) and in ash122 homozygous (ash122, light-green bars) larvae, monitoring H3K36me2 levels at the Ubx, mth, CG6310, wg, tsh, lam and dpr12 genes in T3 discs. At each gene, H3K36me2 was analysed at one or more regions, and for each qPCR, amplicon coordinates are indicated as distance in kb from the transcription start site, see also Table S5. For both genotypes, bars show ChIP signals from three independent ChIP reactions that were performed on three independently prepared batches of chromatin and are presented as a percentage of input chromatin precipitated at each region; dots show individual experimental results and error bars show standard deviation.