X cite series 120q lamp

Cells were seeded on glass coverslips (#1.5 thickness; ~ 170 µm, VWR). At the end of the treatment, the cells were washed twice with PBS, fixed by a 15 min incubation with 2% paraformaldehyde (PFA) in PBS and washed three times. The cells were then permeabilized 5 min with 0.2% Triton X-100 in PBS and washed three times with PBS. The coverslips were incubated 10 min in blocking buffer consisting in PBS 0.1% Tween-20 (PBS-T) containing 5% bovine serum albumin (BSA). The coverslips were incubated for 75 min with the primary antibodies diluted in blocking buffer (mouse anti-γH2AX antibody at 1/1000 and rabbit anti-53BP1 at 1/800), washed four times in PBS-T and then incubated 45 min with the secondary antibodies diluted in blocking buffer, washed four times in PBS-T and twice in PBS, incubated 15 min with 2 µg/mL DAPI (4′,6-diamidino-2-phenylindole) in PBS, washed twice with PBS, dipped in double-distilled water and mounted in VectaShield on a glass slide. Pictures were acquired using an Olympus IX73 microscope fitted with a 40 x UPlanFLN objective (Olympus), a X-Cite Series 120Q lamp (Lumen dynamics), a DP26 camera (Olympus) and using the adequate filters set. The white scale bars on each picture represent 10 µm.