FIG 6.

EV-D68 infection disturbs the spontaneous neuronal activity of the neuronal network in vitro. Measuring the effects of EV-D68 infection and replication on spontaneous neuronal activity by microelectrode array (MEA) recordings. (A) The experimental setup. Cortical cells were cultured until days in vitro (DIV) 7. Prior to mock- or EV-D68-inoculation (MOI of 1, shown in orange), baseline spontaneous neuronal activity was measured (shown in green). At 1 h postinoculation (hpi) the cortical cultures were carefully washed (shown as blue dotted line). At 2, 4, 8, 24, and 48 hpi the spontaneous neuronal activity was measured (shown in pink). (B) The graphical illustration of the percentage of living cells in the neural network of a triplicate of three independent experiments (left graph). The graphical illustration shows the number of neurons expressing dsRNA over time. 10 fields of 100 cells spread over three independent experiments were analyzed (right graph). (C and D) The graphical illustration of the MEA recordings of 16 wells of each condition. (C) The number of spikes is shown as percentage change relative to the control in time postinoculation (hpi). (D) The number of bursts is shown as percentage change relative to the control in time postinoculation (hpi). All data shown in blue are EV-D68 strains that use a carbohydrate receptor for viral entry; All data shown in red are EV-D68 strains that mainly bind to heparan sulfate for viral entry. Ns indicates not significant; * represents P ≤ 0.05; ** represents P ≤ 0.01; *** represents P ≤ 0.001; **** represents P ≤ 0.0001.