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. 2012 Sep 1;23(17):3450–3460. doi: 10.1091/mbc.E11-12-1009

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© 2012 Velichko et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 1: — Hyperthermia induces the phosphorylation of histone variant H2AX at Ser-139 in human cells. (A) Immunofluorescence analysis of γH2AX in control (untreated) human mcf-7 cells and cells that were heat-stressed at different temperatures (42, 44, and 45.5°C for 30 min). The DNA was stained with DAPI. Scale bar: 20 μm. (B) Western blot analysis of γH2AX in the nuclei of control (untreated, C) and heat-shocked (45.5°C, 30 min; HS) mcf-7 cells. Lamin B1 was used as a loading control. (C) Immunofluorescence analysis of heat-treated (45.5°C, 30 min) mcf-7 cells performed using a Leica SP2 confocal laser scanning microscope (scale bar: 20 μm). Enlarged cells with two different patterns of γH2AX distribution are shown on the right (scale bar: 5 μm). The DNA was stained with To-Pro 3 iodide fluorescent dye.