Fig. 5

TGF-β increases NHEJ-mediated DNA repair facilitating proliferation in mCFs which can be rescued upon TET3 overexpression. A mCFs integrated with a DR-GFP HR reporter substrate were transfected in indicated conditions in combination with ISce-1, and analyzed for change in HR efficiency by scoring % of GFP/RFP double-positive cells using flow cytometry. B mCFs integrated with a pLCN-DSB NHEJ reporter substrate were transfected in indicated conditions in combination with ISce-1 and analyzed for change in NHEJ efficiency by scoring % of GFP/RFP double-positive cells using flow cytometry. C Representative graph shows the effect of TGF-β on cell number in control, TET3 knockdown and overexpressed mCFs over 7 days. D Representative graph shows the effect on cell proliferation measured by MTT assay in mCFs pre-treated with TGF-β in control, TET3 knockdown and overexpressed cells. E and F DNA repair constructs showing HR and NHEJ repair efficiency in mCFs pre-treated with in control, TET3 knockdown and TET3 overexpression conditions. G Graph represents BrdU incorporation in non-denaturing conditions, a direct readout for DNA end resection pre-treated with TGF-β in control, and TET3 overexpressed mCFs cells. H Annexin V assay showing change in apoptosis in TGF-β-treated mCFs with or without TET3 overexpression. Statistical significance was calculated using one-way ANOVA. n.s. represents non-significant and P values correspond to *p ≤ 0. 05, **p ≤ 0. 01