Figure 1.

FmKmt1 is involved in H3K9me3 in F. mangiferae. (A) Graphical representation of the domain structure of Neurospora crassa DIM-5 and orthologs in other fungal species including F. mangiferae (FmWT). The conserved pre-Set domain, SET domain, and post-SET domain are indicated; InterPro accession numbers are shown in the domain description (B) For FmKMT1 expression analysis, FmWT, Δfmkmt1, and Δfmkmt1/FmKMT1^Ces were grown on solid CM for 3 days at 30°C. For the determination of transcript levels, RNA was extracted and transcribed into cDNA prior to RT-qPCR. The FmWT FmKMT1 expression was arbitrarily set to 1. Mean values and standard deviations are shown. n.d., not detected; RE, relative expression. (C) Western blot analysis of FmWT, Δfmkmt1, Δfmkmt1/FmKMT1^Ces, and FmH3K9R using two anti-H3K9me3-specific antibodies (AB, Abcam; AM, Active Motif), and one anti-H3K9ac-specific antibody. For referencing one anti-H3-specific antibody was used. Indicated strains were grown in liquid ICI medium supplemented with 120 mM NaNO₃ for 4 days and subsequently total proteins were extracted. Roughly, 50 μg of total protein extracts were separated on a SDS gel prior to western blotting. For quantification, a densitometric analysis was performed and the respective wild-type strain was arbitrarily set to 100%. n.d., not detected via Image J analysis tool.