Aq132f

The cells (5 mL) were fixed with 20 μL of Schaudinn’s fixative (saturated HgCl₂: ethanol, 2:1). Then 10 μL of fixed cells were uniformly spread on a poly-L-lysine-coated coverslip. The cells were washed using PBST (0.05% Triton X-100) for 10 min. The cells were blocked using a blocking solution (3% BSA, 10% normal goat serum, and 0.05% Triton X-100 in PBS) for 1 h at room temperature (RT). They were then incubated overnight with HA antibody (1:500 dilution, #3724S, CST, Danvers, MA, USA), γH2AX (1:200, Clone 2F3, BioLegend, USA), and H3K56ac (1:500 dilution; AB_2661786, Active Motif, Carlsbad, CA, USA) at 4 ℃ overnight. The samples were washed three times with PBST (0.05% Triton X-100) and incubated with FITC-conjugated anti-rabbit IgG antibody (1:1000, AQ132F, Millipore, Billerica, MA, USA) or TRITC-conjugated anti-rabbit IgG antibody (dilution ratio of 1:500, AP192R, Millipore, Billerica, MA, USA) for one hour at RT. The samples were stained with 1 μg/mL DAPI for 15 min and observed using a Delta Vision Elite deconvolution microscope system (Applied Precision/GE Healthcare, Boston, Massachusetts, USA).