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. 2013 Dec 12;42(5):3089–3103. doi: 10.1093/nar/gkt1260

Figure 1.

Figure 1.

TDP1 promotes the repair of alkylation-induced base damage in human cells. (a) Human MRC5 cells were infected with lentivirus particles containing non-targeting control shRNA ‘WT’ or shRNA against human TDP1 ‘TDP1KD’. Cell lysates were fractionated by SDS–PAGE and analyzed by immunoblotting using anti-TDP1 (Eurogentec), anti-Top1 (Santa Cruz) or anti-actin (Sigma) immunoblotting. (b) WT or TDP1KD MRC5 cells were incubated with the indicated concentrations of camptothecin ‘CPT’ for 60 min at 37°C and the ability of single cells to form macroscopic colonies was determined by dividing the average number of colonies on treated plates by the average number of colonies on untreated plates and presented as percentage survival. Data are the mean ± s.e.m. of three biological replicates. (c) Survival was determined in WT and TDP1KD MRC5 cells following exposure to the indicated doses of the alkylating agent methyl methanesulfonate (MMS) for 15 min at 37°C as described in (a). Data are the mean ± s.e.m. of three biological replicates. Asterisks denote statistically significant difference between WT and TDP1KD (P = 0.04, 0.001, 0.02, 0.002 for MMS doses 0.1, 0.25, 0.5 and 0.75 μg/ml, respectively; t-test).