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. 2024 Dec 30;15:10889. doi: 10.1038/s41467-024-55261-3

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 4 — a Evaluation of the effects of ALCAM and DSG-2 knockout (mixed knockout population) on HAdV-C5/B7-GFP attachment and internalization. HAdV-C5/B7-GFP at an MOI of 20 is incubated with cells on ice for 1 h in the absence (attachment assay) or presence (internalization assay) of a follow-up incubation at 37 °C for 45 min. L3 genomic DNA of attached or internalized viruses is quantified by qPCR. Evaluation of the attachment and internalization of HAdV-C5/B7-GFP (b) and clinical HAdV-B7 (c) in non-targeting sgRNA mock cells and ALCAM^-/- cells. d Co-IP analysis of cell lysates from HEK-293T cells co-transfected with ALCAM-myc and HAdV7-fiber-HA overexpression plasmids. e Co-IP analysis of the interaction between ALCAM-myc and different domains of fiber protein, including the full-length fiber-HA, shaft-HA, knob-HA, tail-shaft-HA, or shaft-knob-HA. f Co-IP analysis of the interaction between ALCAM-C-domain-myc or ALCAM-V-domain-myc and HAdV7-fiber-HA, shaft-HA, knob-HA, tail-shaft-HA or shaft-knob-HA. For (e–f), immunoprecipitation is conducted using anti-myc beads. For (d–f), WCL, whole cell lysate. The experiment is repeated three times independently and similar results are obtained. For (a–c), data are presented as mean ± SD (n = 3) from three independent biological replicates. The significant difference between non-targeting and knockout groups is analyzed by two-tailed unpaired Student’s t test. Source data are provided as a Source Data file.