Anti atrx sc 15408

Western blotting and IF-FISH were carried out as previously described39 (link)52 (link). For IF-FISH, the cells were fixed with 4% paraformaldehyde (Sigma-Aldrich, P6148), and then permeabilized with permeabilization buffer (0.5% triton X-100, 20 mM HEPES, 50 mM NaCl, 3 mM MgCl₂, 300 mM sucrose)(Sigma-Aldrich,USA), blocked with 5% goat serum (Gibco, 16210–072) and incubated with primary and secondary antibodies. After incubation, the cells were fixed with 4% paraformaldehyde, then washed with. PBS and dehydrated with ethanol, followed by hybridization with a PNA-TelC-FITC probe (F1002,1:500,Panagene) and FITC-labeled (TTAGGG) peptide nucleic acid (PNA) probe (Panagene, Korea). Figs 1B and 2A used TTAGGG probe and other FISH analysis was performed by telomeric probe CCCTAA. More than 300 cells were counted in each cell line for APB scoring. The following antibodies and their working concentrations were used for Western blotting: anti-Flag (F7425, 1:8000, Sigma-Aldrich); anti-DAXX (sc-7152, 1:400, Santa Cruz Biotechnology); anti-ATRX (sc-15408, 1:400, Santa Cruz Biotechnology); anti-GAPDH (M20006M, 1:6000, Abmart). The following antibodies and their working concentrations were used for IF-FISH: anti-γH2AX (05–636, 1:500, EMD Millipore) and anti-PML (sc-966, 1:400, Santa Cruz Biotechnology).