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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. 5 — a Analysis of the role of ALCAM-EndoA3 axis in HAdV-B7 (clinical isolate) internalization. HAdV-B7 L3 DNA is quantified in non-targeting, EndoA3 knockout-1, EndoA3 knockout-2, ALCAM-/-, Endo A3/ALCAM dual knockout-1 and Endo A3/ALCAM dual knockout-2 cells at 48 h post infection. b Analysis of the individual and combinatorial effects of DSG-2 and ALCAM depletion on clinical HAdV-B7 infection. HAdV-B7 L3 DNA is quantified in non-targeting, ALCAM knockdown, DSG-2 knockout-2, DSG-2 knockout/ALCAM knockdown cells at 48 h post infection of HAdV-B7 infection. c, d Analysis of the interaction between ALCAM (ligand) and HAdV-B7 knob domain (analyte) using SPR with Biacore 8 K. c Association and dissociation curves. d Curve-fitting using a steady-state affinity method with constant Rmax analysis. e Summary of the determined K_D values for different partner proteins. Evaluation of the effects of fiber knob (f) and ALCAM (g) proteins on blocking HAdV-B7 infection in wide-type HEK-293A cells. f HEK-293A cells are pre-treated with fiber knob protein for 30 min and then incubated with HAdV-B7. g HAdV-B7 is pre-treated with ALCAM protein and then supplemented into HEK-293A cells. For (f, g), HAdV-B7 fiber DNA is quantified by qPCR at 45 min after incubation. For (a, b and f, g), data are presented as mean ± SD (n ≥ 3) from independent biological replicates. The significant difference is analyzed by two-tailed unpaired Student’s t test. For (a), the p values between on-targeting,^- EndoA3 knockout-1, EndoA3 knockout-2, Endo A3/ALCAM dual knockout-1 and Endo A3/ALCAM dual knockout-2 cells cell are 5e-8, 1e-7, 8e-6, and 8e-6 respectively. Source data are provided as a Source Data file.