Figure 4. Pol H and Pol K gene silencing triggers H2AX phosphorylation in HeLa-MYC cells.

HeLa cells containing B-DNA or MYC-DNA were transfected with control siRNA (Luciferase), two independent Pol κ siRNAs (siκ1 and siκ2) or two independent Pol η siRNAs (siη1 and siη2). Quantification of γ-H2AX-positive cells in the population of each cell line was performed by Flow Cytometry Analysis as described in Materials and Methods. An example is given in (A) where a scatter plot is presented with γ-H2AX intensity on the y axis and propidium iodide intensity on the x axis for the control B-DNA cells and the MYC-DNA cells depleted for Pol κ. The cell populations used for the measurement of the γ-H2AX-positive cells were enclosed by rectangles. (B) Quantification of the FACS analysis were performed as shown in (A) with the cell lines transfected with the indicated siRNAs. The error bars indicate standard deviations of three independent experiments