Protein kinase CK2 localizes to sites of DNA double-strand break regulating the cellular response to DNA damage
1 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark
2 Department of Radiation Oncology, University of Texas Southwestern Medical Center, Dallas, TX, USA
BMC Molecular Biology 2012, 13:7 doi:10.1186/1471-2199-13-7Published: 9 March 2012
Additional file 1:
Figure S1. 53BP1 focus formation in cells treated with NCS. Cells were transfected with scramble siRNA (si-Scr), CK2α or -α'-siRNA for 72 hours. Where indicated, 0.5 μg/ml neocarzinostatin (NCS) was added to the medium in the last 24 hours of incubation. Fixed cells were subsequently labeled with anti-53BP1 antibody and with a FITC-conjugated secondary antibody. Nuclei were visualized by DAPI staining.
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Additional file 2:
Figure S2. γ-H2AX focus formation in cells treated with cisPT or exposed to UV irradiation. Cells were treated essentially as described in Figure 4D. After treatment, fixed cells were stained with anti-γ-H2AX and subsequently with a FITC-conjugated secondary antibody for revealing the presence of foci of DNA damage. Nuclei were visualized by DAPI staining.
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Additional file 3:
Figure S3. in situ PLA reveals interaction between DNA-PK and histone H3 in cells treated with NCS and expressing CK2. A. Association between DNA-PKcs and histone H3 was investigated by in situ PLA in M059K cells treated as indicated in the figure and exposed to 0.5 μg/ml NCS for 1 hour. The molecular interaction is indicated by the presence of distinct red fluorescent spots in the cell nuclei. Control indicates cells treated with si-Scr and NCS and stained with the secondary antibodies after fixation. B. Quantification of the number of positive signals/cell was performed by computer-assisted image analysis. Mean values +/- SD from three independent experiments are shown. *P < 0.0001 denotes statistically significant difference between cell populations treated with si-Scr and siRNAs against CK2α', respectively.
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