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. Author manuscript; available in PMC: 2017 Nov 15.
Published in final edited form as: Nat Genet. 2017 May 15;49(7):1005–1014. doi: 10.1038/ng.3866

Fig. 3. Ectopic expression of PGBD5 in human cells leads to oncogenic transformation both in vitro and in vivo.

Fig. 3

(a) Schematic for testing transforming activity of PGBD5. (b) Relative PGBD5 mRNA expression measured by quantitative RT-PCR in normal mouse tissues (brain, liver, spleen and kidney), as compared to human tumor cell lines (rhabdoid G401, neuroblastoma LAN1 and SK-N-FI, medulloblastoma UW-228 cells), primary human rhabdoid tumors (PAKHTL, PARRCL, PASYNF, PATBLF), and BJ and RPE cells stably transduced with GFP-PGBD5 and GFP. Error bars represent standard deviations of 3 independent measurements. (c) Representative images of GFP-PGBD5-transduced RPE cells grown in semisolid media after 10 days of culture, as compared to control GFP-transduced cells. (d) Number of refractile foci formed in monolayer cultures of RPE and BJ cells expressing GFP-PGBD5 or GFP, as compared to non-transduced cells (p = 3.6 × 10-5 and 3.9 × 10-4 for GFP-PGBD5 vs. GFP for BJ and RPE cells, respectively). (e) Expression of T. ni GFP-PiggyBac does not lead to the formation of anchorage independent foci in monolayer culture (* p = 3.49 × 10-5 for GFP-PGBD5 vs. T. ni GFP-PiggyBac). Error bars represent standard deviations of 3 independent experiments. (f) Kaplan-Meier analysis of tumor-free survival of mice with subcutaneous xenografts of RPE cells expressing GFP-PGBD5 or GFP control, as compared to non-transduced cells or cells expressing SV40 large T antigen (LTA) and HRAS (n = 10 mice per group, p < 0.0001 by log-rank test). (g) Representative photographs (from left) of mice with shaved flank harboring RPE xenografts (scale bar = 1 cm). Tumor excised from mouse harboring GFP-PGBD5 expressing tumor (scale bar = 1 cm). Photomicrograph of GFP-PGBD5 expressing tumor (top to bottom: hematoxylin and eosin stain, vimentin, and cytokeratin, scale bar = 1 mm).