Fig. 1. Identification of ALCAM as a host factor of HAdV-C5/B7-GFP through SfCRISPR screen in HEK-293T cells.

a Analysis of SCAP-associated HAdVs. The colors are blue for bacteria, pink for atypical pathogens and orange for viruses. The data are extracted from a previous publication⁸ and re-analyzed and the figure re-drawn. b Analysis of DSG-2 transcriptomic expression in the blood samples from SCAP patients with different pathogen infections. FPKM, fragments per kilobase of exon model per million mapped fragments. N.A., no available identifiable pathogen information. Others, all pathogens excluding HAdV. All, all analyzed SCAP patients. Data are presented as mean ± standard deviation (SD) (n ≥ 3) from different patients. Statistical analyses are performed between HAdV group and other groups using two-tailed unpaired Student’s t test. c Construction of HAdV-C5/B7-GFP and evaluation of its infection efficiency in HEK-293A, in comparison with HAdV-C5. Scale bar, 100 μm. The experiment is repeated three times independently and similar results are obtained. d Venn plot showing the overlap of cell surface proteins as defined by each public database. SfCRISPR-v2 contains the union set of these proteins. e Flow chart showing the screening procedure of GFP-negative selection in HEK-293T cells. Bubble plot showing the screening results using SfCRISPR-v1 (f) and -v2 (g) libraries. The compared groups are test and mock infections. Significance of enrichment is calculated using MAGeCK and candidate hits with FDR cut-offs of less than 0.01 are displayed. h Evaluation of the effects of ALCAM, CADM1 and DSG-2 knockout on HAdV-C5/B7-GFP infection in HEK-293T cells by flow cytometry. Data are presented as mean ± SD (n = 3) from three independent biological replicates. The significant difference between non-targeting sgRNA and knockout groups is determined using two-tailed Student’s t test and the P values are shown. The p values between nontarget and DSG-2 knockout-1 is 4e-5. Source data are provided as a Source Data file.