Psg5 fer

pSG5-FER (human FER tyrosine kinase) plasmid was purchased from Addgene (Cambridge, MA, USA) and propagated in E. coli. Plasmids were harvested using QIAGEN Giga-prep kits (Valencia, CA, USA) as per manufacturer’s instructions. The original pSG5-FER plasmid was a gift from Nora Heisterkamp deposited in Addgene DNA repository (Addgene plasmid # 30191). The expression cassette encodes human FER (hFER), which is a non-receptor tyrosine kinase located on human chromosome 5q21 (Gene ID 2241; HPRD 01491; NM_005246). An empty plasmid -pcDNA3.1 (Promega, Madison, WI, USA) containing similar backbone structure without the FER expression open reading frame was used as a control. All plasmids of DNA were stored in 10 mM Tris–1 mM EDTA buffer (pH 8.0) at − 20 °C until the day of gene delivery. Luciferase plasmid PGL-4 (Promega, Madison, WI, USA) with expression being driven by SV40 promoter as in pSG5-FER plasmid was used for reporter gene assays. Protocols for the use of recombinant DNA technology required prior approval from the University of Michigan Institutional Biosafety Committee (IBC) Protocol IBCA00000315.

pSG5-FER (human FER tyrosine kinase) plasmid was purchased from Addgene (Cambridge, MA) and propagated in E. coli. Plasmids were then harvested using QIAGEN Giga-prep kits (Valencia, CA)(54 (link)) per manufacturer’s instructions. Original pSG5-FER plasmid was a gift from Nora Heisterkamp deposited in Addgene DNA repository (Addgene plasmid # 30191)(54 (link)). The plasmid contains an insert that encodes human FER (hFER) which is a non-receptor tyrosine kinase located on human chromosome 5q21 (Gene ID 2241; HPRD 01491; NM_005246). Two separate plasmids containing the α and β subunits of the Na⁺/K⁺-ATPase were a gift from David A. Dean (University of Rochester, NY) and were harvested in similar fashion as described above. A luciferase reporter gene plasmid (Promega, Madison, WI) was used as a control for effects of electroporation process over results, containing similar promoter sequence of the FER plasmid (SV40). Similarly an empty plasmid (pcDNA3) was used for same purposes in survival curve experiments. All plasmids of DNA were stored in 10 mM Tris- 1 mM EDTA buffer (pH 8.0) at −20°C until the day of gene delivery. Protocols for the use of recombinant DNA technology required prior approval from the University of Michigan Institutional Biosafety Committee.