Ic86621

FLAG-Ku70 plasmid was obtained from Shigemi Matsuyama, Case Western Reserve University, Cleveland as a kind gift. FLAG-p65 Plasmid was a kind gift from Katerina Gurova RPCI, Buffalo, NY. Inhibitor of DNA-PK, IC86621; anti-FLAG antibody and M2 affinity- and protein A/G beads were procured from Sigma, USA. DNA-PKc, p65/RelA and PARP-1 antibodies were purchased form Cell signaling Technology, USA. DNA-PKc antibody was also obtained from Kamiya Medical, USA. The anti-β-actin monoclonal antibody was obtained from Abcam, USA. siRNAs were bought from either Eurogenetic or Santacruz biotech, USA. The anti-his₆ antibody was purchased from Biobhatarti life sciences, India. Andrographolide purified (>95% purity) from leaves of Andrographis paniculata was a kind gift from Vinod Kumar Nelson.

DNA-PK Inhibitors: NU7026 (Cat. # S2893) and NU7441 (Cat. # S2638) from Selleckchem, while IC86621 (Cat. # 404009-40-1) from Sigma-Aldrich, was purchased. Stock solutions were made in DMSO at 5 mM concentration. DNA-PKcs shRNA vectors (Cat # sc-35200-SH) were purchased from Santa Cruz Biotech. In some experiments, we also used the lentiviral vector pHR’Sin-Puro with shRNA, which carry puromycin resistance gene and expresses shRNA through H1 Pol-III promoter. The shRNAs to DNA-PKcs (5′-GAACACTTGTACCAGTGTT-3′) or (5′-GATCGCACCTTACTCTGTT-3′) and scrambled control (5′-TTGATGCACTTACTAGATTAC-3′), were cloned. A total of 4 to 6 μg of VSV-G pseudotyped lentiviral vectors carrying specific or control shRNA were used for knockdown.