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Beads conjugated with PtdIns(4,5)P2 and PtdIns(3)P were used as control and beads (from Echelon) with no lipid conjugates was also included in the experiments. The beads were then mixed with SDS-gel-loading dye and boiled for 5 min. The samples were then resolved on gradient SDS-...
SEVs were isolated as previously described.72 Briefly, cells were seeded into T75 culture flasks and cultured until they reached 70~80% confluency. Next, the cells were incubated in the sEVs-free medium. After 24 hours, the cell supernatant was harvested and centrifuged at 300 × g ...
scitamineum sporidia toward various stressful conditions, including oxidative stress (H2O2), osmotic stress (NaCl), and cell wall stress (SDS). We noticed that a hypersensitivity response to H2O2 resulted from SsAGC1 deletion, specifically in cultures grown on PDA or MM-N but not in the ...
This activity was normalized to the protein level determined by BCA assay (Thermo Scientific) after spinning the remaining samples and bead beating of the resulting pellets in 400 μl 1% SDS, 6 m urea in 25 mm Tris (pH 6.8). The trehalase activity per protein content of the pkaas and y...
Immunoblots were performed utilizing lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1mM EGTA, 1 mM Beta-Glycerol-Phosphate, 2.5 mM Sodium-Pyrophosphate, 1% Triton-100X, 0.1% SDS) complemented with fresh protease (Sigma, 11697498001) and phosphatase inhibitors (Sigma, P0044-5ML...