Fig. 3

Validation of the effects of ICAM-1, RAB5C, and OLFML3 on RV infection. a, b Time-dependent viral replication of RV-B14 in mock and knockout cells, as determined by viral loads in medium supernatant (a) and cell lysate (b). Cells are infected with RV-B14 at an MOI of 2. Viral RNA in cell lysates (b) is normalized to RPLP0 expression. Significant difference between test groups and non-targeting sgRNA group is determined using two-way ANOVA with Dunnett’s multiple comparisons test. c, d Rescued susceptibility of knockout cells to the infection of RV-B14 (c) and RV-A16 (d) by overexpression of RAB5C and OLFML3 respectively. e, f Rescued replication of RV-B14 (e) and RV-A16 (f) in knockout cells by overexpression of RAB5C and OLFML3 respectively, as determined by viral loads in medium supernatant. g, h Rescued replication of RV-B14 (g) and RV-A16 (h) in knockout cells by overexpression of RAB5C and OLFML3 respectively, as determined by viral loads in cell lysates. Viral RNA in cell lysates is normalized to RPLP0 expression. i Representative images of CPEs induced by clinical RV strain. Scale bar, 100 μm. j Cell viability of mock and knockout cells upon challenge of clinically isolated RV strain. k Phylogenetic analyses of clinical RV strain using MEGA X [47], with VP4 gene as the reference. l, m The effects of gene knockout on the replication of clinical RV strain, as determined by RT-qPCR quantification of viral loads in medium supernatant (l) or cell lysates (m). For m, viral RNA in cell lysates is normalized to RPLP0 expression. For c–j and l, m, analyses are performed at 24 h post infection of RV at an MOI of 2. Significant difference between test groups and non-targeting sgRNA group is determined using two-tailed Student’s t test and the P values are shown. Significant difference between knockout and overexpression rescue groups is determined using two-tailed Student’s t test and the P values are shown above the lines