Figure 6.

EBNA1-iPSCs have elevated DNA damage compared to dbDNA-iPSCs

nhDFs nucleofected with dbDNA and OriP-EBNA1 vectors were assessed for evidence of DNA damage by comet assay. (A) Representative micrographs of nuclear comets (n > 100 images, scale bar represents 100 μm). (B) Graphical representation of data generated with CaspLab of nhDFs transfected with dbDNA and OriP-EBNA1 including head DNA (Bi), tail DNA (Bii), comet length (Biii), and tail moment (Biv) (n > 100 images, error bars represent ±SEM of 3 independent experiments). ∗∗∗∗p < 0.0001, ∗∗p < 0.01, ∗p < 0.05, Student’s paired t test. (C) nhDFs nucleofected with dbDNA-GFP and OriP-EBNA1-GFP vectors were assessed for evidence of DNA damage response by nuclear γH2AX puncta staining, (Ci) GFP⁺ cells were identified and then γH2AX puncta counted for transfected cells. (Cii) Average puncta per cells counted (>200 nuclei per treatment; scale bar represents 100 μm) graphically represented and compared to untreated nhDFs and those treated with 100 μM H₂O₂ as a positive control. (>200 nuclei per treatment, error bars represent ±SEM). ∗∗p < 0.01.