Supplementary Figure 4: UDS and RRS complementation assays by an electroporation system (Neon, Life Technologies). | Nature Protocols

Supplementary Figure 4: UDS and RRS complementation assays by an electroporation system (Neon, Life Technologies).

From: A rapid, comprehensive system for assaying DNA repair activity and cytotoxic effects of DNA-damaging reagents

Supplementary Figure 4

(a) The experimental scheme of complementation assay by electroporation. (b) UDS and (c) RRS assays for normal 48BR and XP-A-patient derived XP15BR cells. RRS was normalised to activity measurement in non-irradiated cells. Bars and error bars represent average fluorescent intensity of quadruplicate wells and standard deviations, respectively. (d, e) Transfection efficiency was confirmed by immunofluorescent staining with anti-V5 tag antibody. Cells were transfected with indicated plasmid by electroporation and then plated on a glass-bottomed 96-well plate. 40 hours after transfection, immunofluorescent staining was performed as described in Supplementary Method 5. Transfection efficiency was calculated as a number of Alexa 488-positive cells using the VTI system. (f) Total cell counts in the UDS assay. Scale bar, 100 μm; w/o, without transfection.

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