pPuro.HRE>GFP was generated using the lentiviral expression plasmid PURO.Cre (Addgene, plasmid no. 17408) as a backbone. Eight repeats of the hypoxia-responsive element (5′-GCCCTACGTGCTGTCTCACACAGC-3′) from the 3′ enhancer region of the human EPO gene were amplified by polymerase chain reaction (PCR), fused to a TATA box/linker (5′-TCTAGAGGGGTATATAATGGAAGCTC-′3), and amplified with flanking restriction enzyme sites (5′ Xho I/3′ Sma I). The constructed fragment, termed HRE, was inserted into PURO.Cre, replacing the original PGK promoter. Subsequently, EGFP was amplified by PCR from pLNT/SFFV hNIS-GFP (85) with flanking Sma I/Kpn I sites and subcloned into the intermediate construct replacing Cre, thereby providing the final construct pPuro.HRE>GFP. Puromycin (Gibco; 1 μg/ml) was used as the selection antibiotic.

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