Human embryonic kidney (HEK) 293T cells (ATCC, CRL-1573) were transfected with the vector plasmid, the lentiviral packaging constructs pCMV.DR8 and pMD2.G (Plasmid #12259, Addgene). The viral supernatants were harvested at 48 h and 72 h after transfection, filtered with 0.4-μm filters and applied to mouse cardiac microvasculature endothelial cell line (mCMVECs) in the presence of 8 µg/mL polybrene (Santa Cruz Biotechnology) for 6-8 h. The efficiency of infection was determined with fluorescence microscopy and successful Klf2 knockdown by Klf2-shRNA or overexpression by Lenti-Klf2 was determined by Western blot and RT-qPCR.

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