To generate pLenti-ACE2-Hygro (Addgene 161758), we amplified human ACE2 (hACE2) from pcDNA3.1-ACE2 (Addgene 1786) and cloned it into a lentiviral transfer pLEX vector carrying the hygromycin resistance gene using Gibson Assembly Master Mix (NEB E2611L). A 2A epitope tag was added to hACE2 at the C-terminus. Huh7.5ACE2 and A549ACE2 cell lines were generated by lentiviral transduction of ACE2. The protocol for lentiviral production was the same as above except we used the common lentiviral pseudotype (VSV-g) using plasmid pMD2.G (Addgene 12259). Transduced cells were selected with hygromycin at 50 µg/mL for Huh7.5ACE2 and 500 µg/mL for A549ACE2 for 10 days before use.

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