CH25H, IFIH1, IFITM1, IFITM2, IFITM3, and LY6E were cloned into pLX304 lentiviral vector with a C-terminal V5 tag. Human ACE2 was cloned into pDEST-mCherry vector with an N-terminal mCherry tag. TMPRSS2 and TMPRSS4 plasmids were used as previously described (18 (link)). Single-guide RNA against CH25H or NPC1 (SI Appendix, Table S1 ) was cloned into lenti-CRISPR_v2 vector (Addgene, #52961). CH25H point mutations were introduced by QuikChange II site-directed mutagenesis (Agilent, #200524) using primers in SI Appendix, Table S1 . GFP-tagged Rab5 and Rab7 constructs were used as reported previously (64 (link)). Codon-optimized SARS-CoV-2 S was a kind gift from Nevan Krogan at the University of California, San Francisco, CA (65 (link)). pCAGGS-SARS-CoV S was a kind gift of Paul Bates at the University of Pennsylvania, Philadelphia, PA (66 (link)). pMIG-WEEV-IRES-GFP plasmid was generated by Zhuoming Liu in the S.P.J.W. laboratory at the Washington University School of Medicine in St. Louis, St. Louis, MO. PM-GFP and VSV-G plasmids were obtained from Addgene (#21213 and #12259, respectively). pCAGGS-FAST-p10 from pteropine orthoreovirus was generated in the Kobayashi laboratory at the Osaka University, Japan (67 (link)). pEGFP-N1 and pCMV-TdTomato were obtained from Clontech.
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