Ab0045

The protein was extracted as described previously [38 (link)]. Briefly, the mycelia were filtered with Mirocloth, blotted dry, and ground into powder in liquid nitrogen using a mortar and pestle. The mycelial powder was then re-suspended in 1 mL RIPA lysis buffer (1%Triton X-100, 1% sodium deoxycholate, 0.1% SDS, (EpiZyme, PC101)) with 10 uL protease inhibitor cocktail (EpiZyme, GRF101), followed by being shaken once each (10 min) for 30 min on ice. The samples were centrifuged under 12,000× g at 4 °C for 20 min, and the supernatant liquids were acquired as total proteins. The immunoblot analysis was performed with 10% SDS-PAGE, followed by GFP (1:10,000, ABways, AB0045), α-H3K18ac (1:3000, Beyotime, AF5617), and α-H3 (1:2000, Cell Signaling Technology, 4499) antibodies.

The mycelia of strains were cultured in liquid PDA under 180 rpm at 28°C for 36 h, washed with ddH₂O, and moved to MM-N medium under 80 rpm at 28°C for 2 h and 5 h. The mycelia were further collected and ground into powder in liquid nitrogen and then resuspended in 1 mL RIPA lysis buffer (EpiZyme, PC101) with 10 μL of a proteinase inhibitor cocktail (EpiZyme, GRF101). The lysates were incubated in ice for 30 min, resuspended with a Vortex-Genie every 10 min, and further centrifuged at 12,000 × g for 20 min at 4°C. The supernatant proteins were harvested. The proteins were analyzed by 10% SDS-PAGE followed by Western blotting with the primary antibodies being anti-GFP (rabbit, 1:5,000, Abways, AB0045), anti-α-H3K18ac (rabbit, 1:3,000, Beyotime, AF5617), anti-α-H3 (rabbit, 1:2,000, Cell Signaling Technology, 4499), and anti-β-tubulin (mouse, 10,000, Engibody, AT0003) and the secondary antibodies being HRP-labeled goat anti-rabbit lgG (H+L) (1:10,000, Abways, AB0101) and HRP-labeled goat anti-mouse lgG (H+L) (1:20,000, HUABIO, HA1006), respectively. Finally, the proteins were detected by an Omni-ECL Femto Light Chemiluminescence Kit (EpiZyme, SQ201) and analyzed by ImageJ.