SCR7, a specific DNA Ligase IV inhibitor, blocks nonhomologous end-joining (NHEJ).

Complementation of SCR7 Inhibition with Puri?ed Ligase IV: Complementation experiment is carried out by adding increasing concentrations of puri?ed Ligase IV/XRCC4 complex (30, 60, and 120 fmol) along with the oligomeric DNA substrates (5' compatible and 5'-5' noncompatible ends) to the SCR7-treatedextracts. Reactions are incubated for 2 h at 25℃. The reaction products are then resolved on 8% denaturing PAGE. The gel is dried and exposed and the signal is detected with a PhosphorImager and analyzed with Multi Gauge (V3.0) software.

SCR7 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[3]. Wild-type, AAVS1TLR HEK293 and mouse NIH3T3 cells are maintained in DMEM supplied with 15% FBS, cells are passaged three times per week. The mouse Burkitt lymphoma cell line, generated from a Burkitt-like mouse lymphoma is maintained in DMEM supplied with 15% FBS, 2 mM HEPES, 2 mM sodium pyruvate, 2 mM L-glutamine, and 1× NAA, beta-mercaptoethanol and passaged four times per week. For puromycin selection, mCherry+ cells are sorted, seeded at 103 cells/well and selected with 3 mg/mL of Puromycin for 2 weeks. Then colonies are counted and single cells are sorted. The SCR7 inhibitor is purchased, 12 h after transfection these cells are maintained in complete medium supplied with 1 mM SCR7 inhibitor until analysis. At SCR7 concentrations of 60 mM and 10 mM, A reduction of transfection efficiency and of cell viability is observed[3].

DMSO: 45 mg/mL (134.57 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween-80+45% Saline: 2 mg/mL (5.98 mM), Sonication is recommended.