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. Author manuscript; available in PMC: 2011 Jul 2.
Published in final edited form as: FEBS Lett. 2010 May 17;584(13):2926–2930. doi: 10.1016/j.febslet.2010.05.017

Figure 3. Acetylation of lysine 36 of H2AX is not altered by DNA damage.

Figure 3

(a) 293T cells expressing Flag-H2AX were irradiated (5 Gy) and chromatin associated histones isolated. H2AX, γH2AX and H2AXK36Ac were detected by western blot. (b) 293T cells expressing Flag-H2AX, Flag-H2AXK36R or Flag-H2AXS139A were irradiated (5Gy). H2AX, γH2AX and H2AXK36Ac were measured by western blot. (c) H2AX−/− MEFs expressing H2AXwt or H2AXK36R were irradiated (2Gy) and allowed to recover for 1hr. Cells were fixed and immunofluorescent staining carried out using anti-53BP1 antibody. (d) Quantitation of 53BP1 foci in cells expressing H2AX or H2AXK36R mutations. Cells were either untreated (0) or irradiated (2Gy) and the number of cells containing >5 foci counted 1h or 5h post-irradiation. (e) 293T cells stably expressing Flag-H2AX were either growing asynchronously (AS) or synchronized at the G1/S boundary, in S phase or G2/M as described in methods. H2AXK36Ac was monitored using AbK36Ac. (f) Western blots in (e) were scanned, and the ratio of K36 acetylation signal to total H2AX signal calculated.