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Inhibition of exchange proteins directly activated by cAMP as a strategy for broad-spectrum antiviral development 1 Results 2 The EPAC-selective inhibitor ESI-09 has broad-spectrum antiviral activity 2 ESI-09 is a potent inhibitor of SARS-CoV-2 infection 3 ESI-09 tropism and EPAC isoform selectivity 3 ESI-09 has a multimodal mechanism of action that impacts virus entry and spread 4 ESI-09 inhibits viral entry and spread via remodeling of actin filaments 6 EPAC-mediated actin remodeling facilitated via Rho GTPases and Arp2/3 complex 6 ESI-09 inhibits VACV replication in murine lungs ex vivo and in vivo 7 Discussion 9 Experimental procedures 11 Viruses and cell lines 11 Reagents 11 Treatment protocol and reporter assays 12 Plaque assays 12 Viability assays 12 Immunoblotting 12 siRNA KD 12 CRISPR/CAS9 KO lines 13 Virus entry/binding assays 13 Lentivirus assay 13 Immunofluorescence staining of VACV and VSV entry 13 Actin staining 13 Syncytia colocalization assay 13 SARS-CoV-2 experiments 13 SARS-CoV-2 immunofluorescence and confocal microscopy 14 Murine explants 14 Human patient explants 14 In vivo challenge experiments 14 Statistical tests 15 Data availability 15 Supporting informatio

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nonfiction