Figure 1 | Scientific Reports

Figure 1

From: PREP1 tumor suppressor protects the late-replicating DNA by controlling its replication timing and symmetry

Figure 1

PREP1 down-regulation affects cell cycle and DNA replication rate of HeLa cells. (A) Immunoblot on a whole cell lysate showing the efficiency of PREP1 down-regulation 48 hrs after transfection with oligonucleotides 607 and 900. Luc siRNA was used as control. Actin was used as loading control. (B) Cell cycle analysis (see Methods). Percent of cells (normalized to the number of cells at the end of BrdU pulse, t = 0) in early-, mid- and late-S phase fractions, measured 4 hrs after the release from BrdU administration, in siLuc- (empty) and siPREP1-(black) transfected cells. The data are the average of three independent experiments. Asterisks indicate significant p-values (Student’s T test): 0.0203 (Early-S), and 0.0172 (Late-S). (C) (Top) Representative DNA fiber showing the incorporation of I-dU (red) and Cl-dU (green) DNA precursors. The mid-point between two red stretches is the replication origin. The length of the red stretches and the time of incubation allow the measurement of the replication rate. (Bottom) Measurement of DNA replication rates on individual DNA fibers (n = 511 and 565 in siLuc and siPREP1 cells, respectively). p-value (Student’s T-test), as indicated.

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