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. 2019 Jul 24;8(1):1076–1085. doi: 10.1080/22221751.2019.1644142

Figure 2.

Figure 2.

VP2 substitution V135I in mouse-adapted EV-A71 (#812MA) is a genetic determinant of neurotropism (A) Side view of a protomeric structure of EV-A71 (3VBS) along the 2-fold symmetry axis (USCF Chimera). The four capsid proteins VP1-4 are depicted in blue, green, red and purple, respectively. A close up view of the EF loop and amino acid V135 of VP2 (in red) is also shown. (B) Scheme of EV-A71 #812 in vitro adaptation. EV-A71 #812 was used to infect neuroblastoma murine N2a cell line (MOI = 1). Three days p.i., supernatant (diluted 1:5 was used to infect a fresh monolayer of N2a cells). At passage 0, 3 and 8, viral RNA was extracted and VP2 was sequenced. (C) Replication kinetics of EV-A71 #812 and #812MA on N2a cells. At the indicated times p.i., N2a cells were harvested and viral levels were determined by means of end-point titration on RD cells. Error bars represent the mean ± SEM of 3 biological replicates. P values were calculated by two-way ANOVA in GraphPad, *P < .05; ns, P > .05.