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. Author manuscript; available in PMC: 2015 Feb 20.
Published in final edited form as: J Cell Sci. 2012 Mar 7;125(0 4):1048–1057. doi: 10.1242/jcs.098392

Fig. 7. The LEM domain in Ankle1 is required for inducing DNA damage response.

Fig. 7

HeLa cells were transfected separately with constructs expressing Ankle1-V5 or the LEM domain-deficient Ankle1b-V5 and seeded as a mixed culture onto cover slips after 24 hours. The plasmid expressing Ankle1-V5 also expresses a CMV-driven GFP-Blasticidin marker gene (GFP-Bsd), the plasmid encoding Ankle1b-V5 contains a Blasticidin gene without GFP fusion (Bsd) (plasmid maps). After 48 hours cells were either treated with Leptomycin or ethanol for three hours, fixed, stained for V5 (red) and γH2A.X (yellow) and imaged on a confocal fluorescence microscope. DNA was stained with DAPI (blue). GFP-Blasticidin is shown in green. Cells marked with asterisks lack GFP signal and are thus expressing Ankle1b; cells expressing Ankle1 (expressing GFP) are marked with arrows. Relative fluorescence intensities of V5 and γH2A.X signals within the nucleus of untransfected, GFP-positive and GFP-negative transfected and Leptomycin-treated cells (n>30 each) were measured and plotted using Graphpad Prism software. Representative results out of three independent experiments are shown.