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Figure 3


Fig. 3. hOGG1 relocalisation to nuclear speckles is not dependent on the recognition of the lesion 8-oxoG. (A) Fpg- and T4-endoV-sensitive sites and single-strand breaks (ssb) were measured by alkaline elution in non-irradiated (NI) and UVA-irradiated cells. Bars represent the average of at least three determinations ± s.d. (B) HeLa cells were transiently transfected with the plasmids encoding hOGG1-GFP or the mutant proteins K249Q-GFP and F319A-GFP. Protein extracts were analysed by western blotting with an anti-hOGG1 antibody. (C) The same protein extracts were assayed for hOGG1 glycosylase activity. S and P indicate the substrate and the product, respectively. (D) Transiently transfected cells were UVA-irradiated and analysed under the confocal microscope. Both mutant proteins K249Q and F319A, as well as the wild-type hOGG1 (green), were able to re-localise into foci co-localising with SC35 (red). Bars, 4 µm.





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