Fig. 1: ISG screen identifies LY6E as a potent CoV restriction factor.
From: LY6E impairs coronavirus fusion and confers immune control of viral disease
a, Duplicate screens depicting HCoV-229E infection (24 h post-infection) in Huh7 cells ectopically expressing ISGs. b, HCoV-229E infection of Huh7 cells expressing LY6E or control vector (empty) (blue: DAPI; green: HCoV-229E N protein; red: TagRFP encoded in SCRPSY vector). c–h, Effect of stable LY6E expression on diverse CoVs. Cells were infected with HCoV-229E (c), HCoV-OC43 (d), MERS-CoV (e), SARS-CoV (f), SARS-CoV-2 (g) or MHV-Gluc (h). RLU, relative light units. i, Uniform manifold approximation and projection (UMAP) visualization of LY6E expression and distribution in primary hAEC cultures from two donors by scRNA-seq. j, Induction of LY6E in mock or infected hAECs from three donors by RNA-seq. k, SARS-CoV-2 infection of LY6E or control vector (FLuc) Huh7.5 cells co-expressing human ACE2 (blue: DAPI; green: SARS-CoV-2 dsRNA; red: TagRFP encoded in SCRPSY-ACE2 vector). l, Quantification of the dsRNA mean fluorescence intensity (MFI) normalized by the number of nuclei in the image. m, Western blot of wild-type (WT) A549 or two LY6E knockout (KO) clones (numbers 3 and 4). n, HCoV-229E infection of wild-type or LY6E knockout A549. o, Western blot of wild-type or LY6E knockout A549 reconstituted with CRISPR-resistant LY6E (CR LY6E) or control vector (empty). p, HCoV-229E infection of CRISPR-resistant LY6E-reconstituted wild-type or LY6E knockout A549. Data represent the means of independent biological replicates (n = 3 (c, d, f–h, j and l) and n = 4 (e, n and p)). Immunofluorescence images are depicted exemplarily from n = 2 (b) and n = 3 images (k). In l, seven images per condition were acquired and processed. Scale bars: 200 µM (b) and 100 µM (k). Statistical significance was determined by two-tailed unpaired Student’s t-test with Welch’s correction (c, d, f and h), one-tailed Mann–Whitney U-test (e), two-tailed ratio-paired Student’s t-test (g), one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test (l) or two-way ANOVA followed by Šidák’s or Dunnett’s multiple comparisons test (n and p). For the box plots, centre lines indicate mean values, and upper and lower bounds, respectively, indicate maximum and minimum replicate values (j and l). Error bars represent s.d. P values: **P = 0.0088 (c); ***P = 0.0001 (d); *P = 0.0143 (e); *P = 0.0286 (f); *P = 0.0101 (g); ***P = 0.0010 (h); **P = 0.0035, **P = 0.0091 and **P = 0.0057 (left to right in l); **P = 0.0073 and **P = 0.0033 (left to right in n); and **P = 0.0013 and ***P = 0.0001 (left to right in p). NS, not significant.
