Fig. 4: ITGB4 confers susceptibility of ZIKV to HEK-293T cells.

a HEK-293T cells were transfected with plasmids encoding human ITGB4 or ITGA6 for 48 h and infected with ZIKV as described above. Cells were subsequently inoculated for 1 h with the virus-protein mixes on ice. Cells were washed three times with PBS and the virus titers were determined by plaque assay on Vero cells at 48 h after infection. The values in the graph represent the mean ± SD of three independent experiments. ****P < 0.0001 (SZ_SMGC-1, top), **P = 0.0012 (MR766, top), ***P = 0.0007 (MR766, bottom), two-sided, multiple comparisons by One-way ANOVA test. b HEK-293T cells were transfected with plasmids encoding human ITGB4 for 48 h, and then infected with ZIKV that preincubated with the indicated concentrations of soluble ITGB4, ITGB4/ITGA6 or BSA. After incubated with 1 h with virus-protein mixes on ice, cells were washed three times with cold PBS. ZIKV RNA levels were determined by RT-PCR assay. The data were expressed as Mean ± SD of three independent experiments. **P = 0.0032 (left, top), ****P < 0.0001 (left, bottom), **P = 0.0014 (right, top), ****P < 0.0001 (right, bottom), two-sided, multiple comparisons by One-way ANOVA test. c Effects of ITGB4 antibody on ZIKV SZ_SMGC-1 binding. HEK-293Tcells were transfected with plasmids encoding human ITGB4 were treated with the indicated amount of 13H10 antibody or an isotype antibody before incubation with ZIKV. 1 h after incubation on ice, cells were washed, and total RNA was extracted. ZIKV RNA was quantified by RT-PCR. The data were expressed as Mean ± SD of three independent experiments. ***p = 0.0003, *p = 0.0129, two-sided, multiple comparisons by One-way ANOVA test.