Fig. 3: VAMP5 is a pan-coronavirus restriction factor for ESCs.

a–e RT-qPCR analysis of VAMP5 WT and KO clones (n = 2 cell lines) in hESCs influence on viral replication of coronavirus, such as MERS-CoV (a), OC43 (MOI = 1, 24 hpi) (b), NL63 (MOI = 1, 24 hpi) (c), PEDV (MOI = 0.1, 24 hpi) (d) and MHV (MOI = 0.1, 24 hpi) (e). f–j RT-qPCR analysis of viral RNA levels from ZIKV (MOI = 0.1, 24 hpi) (f), VSV (MOI = 0.1, 24 hpi) (g), DENV2 (MOI = 0.1, 24 hpi) (h), IAV (MOI = 0.1, 24 hpi) (i) and EV71 (MOI = 0.1, 24 hpi) (j) infecting VAMP5 WT and KO clones (n = 2 cell lines) in hESCs. k, l Luciferase activity analysis of viral replication from AAV2 (MOI = 0.1, 24 hpi) and VSV-G-pseudotyped HIV-1 lentivirus (LV) (MOI = 0.1, 24 hpi) with luciferase as reporter for viral replication in VAMP5 WT and KO clones (n = 2 cell lines) hESCs. m Representative images of VSV-GFP expression in VAMP5 siNC and siRNA induced knocked down hESCs (MOI = 0.1, 24 hpi) (Scale bars, 100 μm) and quantification of GFP fluorescence signal density. n Graphic representation of VAMP5 inhibiting viral infection in vivo. Nude mice were xenografted with ESCs-H9 cells exposed to PEDV and DENV2 (2 × 10⁵ pfu for one mouse), followed by infection of above viruses at set intervals. Created in BioRender. Wang, K. (https://BioRender.com/w02jplv). o Equal amounts of total RNA were obtained from the ESCs clusters to detect PEDV and DENV2 levels by RT-qPCR. WT, wildtype; KO, knock-out; NC, negative control; LV, lentivirus. Data are represented as mean ± SEM, n = 3 biological replicates in each group. For in vivo experiment (o), each group, n = 5 biological replicates as indicated. Student’s two tailed t test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.