Fig. 1

Induction of thioguanine resistance and loss of HPRT1 expression in isogenic cells expressing DNA transposase PGBD5. a Stable expression of GFP-PGBD5 and control GFP in BJ-hTERT cells, as assessed by Western blotting against GFP; β-actin serves as loading control. b Clonogenic efficiency of BJ-hTERT cells stably expressing GFP (red) and GFP-PGBD5 (blue) as a function of varying thioguanine concentrations upon thioguanine resistance selection. c Representative photographs of resistant colonies stained with Crystal Violet. d Thioguanine selection of both GFP and GFP-PGBD5 expressing cells yields cells that lack HPRT1 activity, as assessed by hypoxanthine-aminopterin-thymidine (HAT) treatment; *** denotes p = 2.2 × 10⁻⁵ and 9.6 × 10⁻⁴ for the comparisons between control and thioguanine selected GFP and GFP-PGBD5, respectively. d Thioguanine selection yields thioguanine-resistant cells, as assessed by cellular ATP luminescence assay of GFP (red) and GFP-PGBD5 (blue) expressing cells, as compared to control cells (gray and black). e Western blot for HPRT1 in BJ-hTERT cells expressing GFP and GFP-PGBD5 upon thioguanine selection; β-actin serves as loading control. All error bars represent standard deviations of 3 biological replicates