FIG. 1.

HMGB1 facilitates Sleeping Beauty (SB)-mediated transposition from the context of an HSV-1 amplicon plasmid. (A) Schematic representation of the colony formation assay. All assays were performed at a minimum of n = 3 per condition. (B) Western blot analysis was conducted with an anti-HMGB1 rabbit polyclonal antibody to confirm the lack of HMGB1 protein expression in the HMGB1-deficient mouse embryonic fibroblast (MEF) cell line. Wild-type MEF cells served as a positive control. (C) HMGB1-deficient MEF cells were cotransfected with pHSVT-Zeo in combination with pHSV-SB and pHSV-HMGB1 at an equivalent mass ratio. pHSVPrPUC served as a negative control and was employed to ensure that equal amounts of DNA were transfected under each condition. After a 14-day antibiotic selection period, the numbers of Zeocin-resistant colonies were enumerated via methylene blue staining. Error bars represent the standard error of the mean and statistical analysis was conducted by Student t test. Values of p are indicated.