Fig. 1: Eaf3 physically interacts with Ash1 in M. oryzae.

a Summary of Ash1-interactors identified by a mass spectrometry assay. Total proteins were extracted from an ASH1-Flag strain and immunoprecipitated with anti-Flag magnetic beads, followed by a mass spectrometry assay. b Gene structure of EAF3 showing tandemly arranged chromodomain (CD)-containing MGG_06716, MRG domain-containing MGG_06717, and EAF3. Integrative genome viewers (IGV) were used to show the expression pattern in the RNA-seq assays of WT, ∆eaf3, ∆mgg_06716 and ∆mgg _06717 strains. c Radial growth and statistical analysis of colony diameters in the indicated strains. Colonies of the indicated strains were grown on a complete medium (CM) for 7 d, then the top side of the colonies was imaged. Values represent means ± standard deviation (SD) from three biological replicates and statistical significance to the WT strain was assessed by a two-tailed Student’s test (**P < 0.01). n.s., no significance. d A Venn diagram showing significant overlap of gene sets among up-regulated genes in the indicated strains. A P-value indicates the significance of the overlaps between two gene sets using two-sided Fisher’s exact test. e Yeast-two-hybrid assays between Eaf3 and Ash1. Full-length and truncated fragments of EAF3 and ASH1 were cloned into pGBKT7 or pGADT7 respectively. The bait and prey plasmids were co-transformed into Y2Hgold yeast strain. Subsequently, co-transformants were grown on basal medium without tryptophan and leucine (SD-WL) and selective medium without tryptophan, leucine, histidine, and adenine (SD-WLHA). AD and BD, plasmid with empty pGADT7 or pGBKT7 used in the experiments. f Co-immunoprecipitation (Co-IP) assays were performed with the strains co-expressing EAF3-GFP and ASH1-Flag. Immunoprecipitation was performed with anti-Flag M2 magnetic beads and western blotting was conducted with anti-GFP and anti-Flag respectively. Two independent experiments were performed with similar results.