Plenti6.3 to v5 dest vector

The pLenti6.3/TO/V5-H-Ras vector was obtained from Dr Shiaw-Yih Lin (Department of Systems Biology, MD Anderson Cancer Center). Briefly, PCMV-H-RAS (V12) was purchased from Invitrogen (Waltham, MA, USA) and subcloned into the pLenti6.3/TO/V5-DEST vector (Invitrogen) based on the commercial protocol of ViraPower TM HiPerform TM T-Rex TM Gateway Expression system (Thermo Fisher Scientific Waltham, MA, USA). pLenti3.3/TR and pLenti6.3/TO/V5-H-Ras were stably integrated into MCF10A cells. Expression of H-Ras was induced by the addition of doxycycline (1 μg/ml) for 48 h. The vector that expresses FLAG-tagged DNA2 has been described.^{33 (link)} DNA2 short hairpin RNA vectors and control short hairpin RNA vectors against luciferase sequences were obtained from Sigma (St Louis, MO, USA; shDNA2#1: 5′-CCGGACCTGGTGTTGGCAGTCAATACTCGAGTATTGACTGCCAACACCAGGTTTTTTTG-3′; shDNA2#2: 5′-CCGGAGTTTGTGATGGGCAATATTTCTCGAGAAATATTGCCCATCACAAACTTTTTTTG-3′). On-target smart pool small interfering RNAs against DNA2 were purchased from Dharmacon Research (Lafayette, CO, USA; #9: 5′-AGACAAGGUUCCAGCGCCA-3′; #10: 5′-UAACAUUGAAGUCGUGAAA-3′ #11: 5′-AAGCACAGGUGUACCGAAA-3′; #12: 5′-GAGUCACAAUCGAAGGAUA-3′). Transfection of plasmids was performed with the FuGENE 6 reagent from Roche (Indianapolis, IN, USA). Transfection of small interfering RNAs was performed with Oligofectamine (Invitrogen).