Figure 8.

Ectopically expressed met-8 can be stimulated by its upstream heterochromatic domains. (A) Schematic depicting two AT-rich DNA regions located on linkage group III (top panel) and linkage group I (bottom panel) of N. crassa genome, respectively. Bars (primer pairs 6–9) above the schematic indicate the regions used by qPCR in ChIP assays. (B) ChIP analyses showing enrichment of H3K9 trimethylation at the AT-rich region 1 or region 2 in wild-type, cul4^KO, ddb1^KO, dcaf26^KO, dim-7^KO and dim-5^KO strains. (C) Schematic showing the loci where the pmet-8(1.0-kb)-5Myc-6His-MET-8::hph cassette integrated into the downstream of AT-rich region 1 or AT-rich region 2. (D) Western blot analyses using antiserum against MET-8 showing protein levels of ectopic myc-met-8 genes integrated into the his-3 locus (his-3-MET-8) and the nearby AT-rich region 1 (AT1–MET-8) or AT-rich region 2 (AT2–MET-8), and endogenous MET-8 levels, respectively. (E) Densitometric analyses from four independent experiments showing the western blot analysis of (D).