Figure 2

ATF3 acts as a negative regulator of the antiviral response. (a) Neuro2a cells were treated with either a control non-targeting siRNA or the Atf3-specific siRNA for 48 h and then infected with JEV (MOI 5). Total proteins from the cell lysates harvested at 24 h pi were Western blotted to establish the ATF3 depletion. (b) Total RNA was isolated from the cells at 24 h pi and qRT PCR performed in triplicates to determine the relative transcript levels for various ISGs or members of UPR pathway. Mean values were used to create the heat map demonstrating the gene-expression profiles using the Gene-E software. (c) Neuro2a cells were treated with either a control non-targeting siRNA or the Atf3-specific siRNA for 48 h and then infected with JEV (MOI 5) for different time points. The experiment was done in triplicates on three different occasions. Relative expression of various genes at different times pi was studied by qRT PCR performed in triplicates for each sample, and the mean is plotted. Transcript level in control siRNA-treated cells is shown by the black bar while that in the Atf3 siRNA-treated cells is shown by the gray bar. The Gapdh levels were used for normalization. The statistical significance of the difference between the data was established by Student’s t-test; *p < 0.05; **p < 0.01; ***p < 0.001.