Pmscv ires mcherry

NK cells were isolated from spleen of C57BL/6 WT mice and NK cell mRNA was purified using RNeasy Mini Kit (QIAGEN). Total cDNA was prepared by reverse transcription and DNA was then amplified using the following primers: XCL1-Forward (BamHI) 5′-GGCCCGGGGATCCATGGATGAGACTTCTCCTCCT-3′ and XCL1-Reverse (XhoI) 5′-CGGCCAACCGGCTCGAGTTACCCAGTCAGGGTTA-3′ for XCL1; CCL5-Forward (BamHI) 5′-GGCCCGGGGATCCATGGATGAAGATCTCTGCAG-3′ and CCL5-Reverse (XhoI) 5′-CGGCCAACCGGCTCGAGCTAGCTCATCTCCAAATA-3′ for CCL5. Both PCR products and the target vector pMSCV-IRES-mCherry (Addgene #52114) were then digested with XhoI and BamHI for 1h at 37°C and purified by gel extraction after agarose gel electrophoresis. Ligation was performed for 1h at room temperature using T4 DNA ligase (NEB). The ligation mix was then transformed into One Shot TOP10 chemically competent bacteria (Thermo Fischer Scientific) and plated on ampicillin containing LB-agar plates. Single colonies were then sequenced and used for plasmid isolation using the High speed Maxi Kit (QIAGEN).