Fig. 3.

Pulsed-field gel analysis of BCM-induced DSBs. Cells were treated with 100 μg/mL BCM for 12 h where indicated. DNA fragments were separated on a 1% agarose gel in 0.5× TBE at 14 °C for 26.4 h at 6 V/cm (200 V) using a 120° included angle with a 2.8- to 26.3-s linear switch time ramp. Intact chromosomes remain in the well, and linearized DNA runs at 3.7 Mb. (A) PFGE. lane1: λ concateners from 0.05 to 1.0 Mb; lane 2: RSW464; lane 3: BCM-treated RSW464; lane 4: RSW919; lane 5: BCM-treated RSW919. (B) Ratios of linearized to intact chromosomes. Ratio linear:cc = amount of linearized chromosomal DNA divided by amount of DNA retained in well. Fold-increase = fold-increase in linear DNA after BCM treatment. (C) UV sensitivity. Dilutions (10²-, 10⁴-, and 10⁶-fold) of fresh overnight cultures of MDS42, RSW455, and RSW919 were exposed to 0 or 40 J/M² UV irradiation, and incubated at 37 °C for 24 h.