Anti 53bp1 antibody

Cells were cultured in 8-well chamber slides at a density of 3 x 10⁴ cells/well overnight and treated with 0.01% MMS for 1 hr. Then, the cells were fixed with 3.5% paraformaldehyde for 30 minutes at room temperature. The fixed cells were incubated in blocking buffer (10% FBS, 0.1% Triton X100 in PBS) for 30 minutes at room temperature and further incubated with anti-γH2AX antibody (1:100, Millipore, 05–636) and anti-53BP1 antibody (1:100, Abcam, ab36823) at 4°C overnight. Alexa Fluor 594-labeled goat anti-mouse IgG (H+L) antibody (1:500, ThermoFisher Scientific, A-11032) and Alexa Fluor 488-labeled goat anti-rabbit IgG (H+L) antibody (1,500, ThermoFisher Scientific, A-11001) were then used to detect γH2AX and 53BP1 foci. ProLong Gold Antifade Mountant with DAPI (ThermoFisher Scientific, P36935) was used to stain nuclei. Images were captured using a Zeiss LSM 780 confocal Microscope, and the intensity of γH2AX and 53BP1 was quantified with ZEN 3.3 (blue edition) in at least 100 cells were measured per cell line.

Acetyl-Histone H4 (Lys8): Millipore, Billerica, MA; Cat # 17–10099
Anti-Histone H4 antibody: Millipore, Billerica, MA; Cat # 05-858R; clone 62-10C-2
Anti-hnRNPA2 antibody: Santa Cruz Biotechnology, Dallas, TX; Cat # sc-32316, clone # DP3B3
Anti- beta actin antibody: Santa Cruz Biotechnology, Dallas, TX; Cat # sc-47778, clone # C4
Anti-53BP1 antibody: Abcam, Cambridge, MA; Cat # ab36823
Anti-gamma H2A.X antibody: Abcam, Cambridge, MA; Cat # ab2893