Goat α rabbit

For GFP-tagged proteins, mouse α-GFP antibody (SC-9996, SantaCruz) was used at 1:1,000 dilution in blocking solution (TBST with 5% milk). Secondary goat α-mouse (170–6516, Biorad) was used at 1:2,000 dilution in blocking solution. For the detection of SkpA, custom made rabbit α-SkpA was used at 1:1,000 dilution in blocking solution. For the detection of phosphorylated MpkB, rabbit α-phospho p44/42 (Cell Signalling Technology) was used at 1:1,000 dilution in TBST with 5% BSA. Goat α-rabbit (Biorad) was used as a secondary antibody for both SkpA and phosphorylated MpkB detection at 1:2,000 dilution in blocking solution.

For GFP-tagged KdmB, mouse α-GFP antibody (Cat# SC-9996, SantaCruz) was used at 1:1,000 dilution in blocking solution (TBST with 5% milk). Secondary goat α-mouse (Cat# 170–6516, Biorad) was used at 1:2,000 dilution in blocking solution. A rabbit polyclonal antibody against a subunit of the SCF complex, SkpA (SconC) was used (1:2,000 dilution in TBST with 5% milk) as a loading control with the Goat α-Rabbit (Bio-Rad, Cat# ab190479; 1:2,000) secondary antibody. For nuclear isolation, approximately 2x10⁶ spores were inoculated into GMM with required supplements for 24 h at 30 °C submerged culture. Buffers and extraction protocol were performed similarly as described previously (Soukup and Keller, 2013 ). Antibodies (1:2,000 dilution in blocking solution) used for histone PTMs were as following; α-H3 (Abcam; AB1791), α-H3K4me3 (Active Motif; 39159), α-H3K9me3 (Abcam; ab8898), α-H3K36me3 (Abcam; ab9050), α-H3K9ac (Merck; 07–352), α-H3K14ac (Merck; 07–353).