Fig. 5: C-terminal of VAMP5 is functionally conserved and responsible for restricting coronavirus infection. | Nature Communications

Fig. 5: C-terminal of VAMP5 is functionally conserved and responsible for restricting coronavirus infection.

From: Vesicle-associated membrane protein 5 is an intrinsic defense factor for embryonic stem cells against coronaviruses

Fig. 5

a Graphic representation of VAMP5 constructs with different region deletion. VAMP5-N contains SNARE domain, and VAMP5-C contains TM domain and CTD. b Representative images of western blot analysis of SARS-CoV-2 spike protein (S) expression impaired by transfecting VAMP5 constructs with different region deletion (MOI = 0.05, 24 hpi). The results are representative of three independent experiments. c Graphic representation of VAMP5 sequence comparison among Homo sapiens, Gorilla gorilla gorilla, Macaca mulatta, Mus musculus and Rattus norvegicus. d–i RT-qPCR analysis of viral RNA levels from SARS-CoV-2 WT (d) and variants like Alpha (e), Gamma (f), Kappa (g), Delta (h) and Omicron (i) infection (MOI = 0.05, 24 hpi) in VAMP5 KO Caco-2-N cells rescued with VAMP5 overexpression from different species. j Table shows the frequency of three SNPs in VAMP5 in several human populations. k RT-qPCR analysis of viral RNA levels of SARS-CoV-2 infecting VAMP5 KO Caco-2-N cells rescued with VAMP5 WT and mutants (MOI = 0.05, 24 hpi). l Western blot analysis of SARS-CoV-2-S expression impaired by VAMP5 WT and mutants in VAMP5 KO Caco-2-N cells. The results are representative of three independent experiments. SNARE, soluble N-ethylmaleimide-sensitive factor attachment protein receptor; TM, transmembrane domain; CTD, C-terminal domain; WT, wild-type; KO, knock-out; Ctrl, control; Hom, homo sapiens; Gor, Gorilla; Mac, macaca mulatta; Mus, mus musculus; Rat, rattus norvegicus. Data are represented as mean ± SEM, n = 3 biological replicates in each group (d–k). Student’s two tailed t test was used. *p < 0.05, **p < 0.01.

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