Figure 3
Functional characterization of the pre-existing POLD1 variants in DLD-1. (A) Representative results of constitutive protein levels (n = 3) in POLD1+/+, POLD1R689W/- and POLD1+/- cells as well as protein levels 120 h after siPOLD1 transfection in POLD1+/+ cells by immunoblotting. β-Actin served as loading control and original immunoblots are displayed in Figure S2. (B) Quantification of the elongation rate, (C) 2nd pulse origins and (D) stalled replication forks upon treatment of POLD1+/+ and POLD1R689W/- cells with IR or HU via DNA fiber assay. Error bars represent mean ± SEM of three independent experiments with at least 100 DNA fibers analyzed in each experiment. Using a two-tailed, unpaired Student’s t test, statistical significance could not be reached.
