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. Author manuscript; available in PMC: 2021 Feb 28.
Published in final edited form as: Nat Microbiol. 2020 Jul 23;5(11):1330–1339. doi: 10.1038/s41564-020-0769-y

Extended Data Fig. 1. The antiviral activity of LY6E is conserved, independent of CoV receptors, and extends to HCV.

Extended Data Fig. 1

a, Duplicate screens depicting HCoV-229E infection (48 hours post-infection) in Huh7 cells expressing ISGs. b, SARS-CoV-2 infection of Huh7.5 cells expressing LY6E or a control (FLuc). c, HCoV-229E infection of Huh7.5 cells expressing vector control or human (H. sapiens), camel (C. dromedarius), bat (P. alecto), mouse (M. musculus), or rhesus (M. mulatta) LY6E orthologues. d, Stable LY6E or empty vector expressing Huh7.5 cells infected with MERS-CoV. e-g, UMAP visualization of CD13 (e) DPP4 (f), and ACE2 (g) expression and distribution among distinct cell types in hAEC cultures from 2 donors by scRNA-seq. h, SARS-CoV-2 infection of LY6E or control vector (FLuc) Huh7.5 cells co-expressing control vector (empty). Blue: DAPI, green: SARS-CoV-2 dsRNA, red: TagRFP encoded in SCRPSY-ACE2 vector. i, Western blot of Huh7.5 expressing LY6E or FLuc and empty vector or ACE2. j-k, Stable LY6E or empty vector expressing Huh7 cells were assessed for cell surface expression of CD13 (j) and DPP4 (k). l, Stable Huh7.5 cells expressing either LY6E or empty vector infected with HCoV-229E, HCV, CHIKV, hPIV-3, RSV, SINV, VEEV, WNV, ZIKV, YFV, and DENV. m, HCoV-229E-Rluc infection of naïve or stably transduced Huh7 cells expressing empty vector control or LY6/uPAR family members. n, HCoV-229E-Rluc infection of Huh7 cells expressing control vector (FLuc), LY6E WT, LY6E HA or specific block mutants9. In h, scale bars are 100 μM. Immunofluorescence and western blot images are depicted exemplarily from n=3 (h) and n=3 (i). Data depicts mean of independent biological replicates, n=3 (b-d,j-n). Statistical significance was determined by two-tailed unpaired student’s t-test with Welch’s correction (b,j-k), ordinary one-way ANOVA with Dunnett`s correction for multiple comparison (c-d, m-n), two-way ANOVA followed by Sidak’s multiple comparisons test (l). Error bars: SD (b-d,j-n). P values (left-to-right): b, ** p=0.0098; c, **** p=<0.0001, ** p=0.0030; d, * p=0.0118, ** p=0.0013; l, **** p=>0.0001, ** p=0.0095; m, **** p=<0.0001; n, **** p=<0.0001, <0.0001, *** p=0.0005, **** p=<0.0001, *** p=0.0003, **** p=<0.0001, <0.0001, <0.0001, *** p=0.0001, **** p=<0.0001.