Figure 3

Validation of microarray results confirms that JunD has both activatory and repressive roles in controlling gene expression linked with oxidative stress. Validation of differentially expressed genes were carried out using four biological replicates with three technical amplification replicates per siRNA type for the unstimulated (A) and eight hour LPS stimulated (B) data sets. Relative gene expression was normalised to Hprt and used to generate fold change values. *P<0.05; **P<0.01;***P<0.001 using a two-tailed unpaired t-test to detect statistically significant differences between the siRNA groups. Error bars represent standard error of the mean. Confirmation of the differential expression of Jund between WKY and WKY.LCrgn2 BMDMs (C) and key JunD targets; Arg1 (D), Cdo1 (E) and Mt2a (F), influenced by Jund siRNA knockdown with >2 fold change in expression that were also differentially expressed between WKY and WKY.LCrgn2 BMDMs over a timecourse of LPS stimulation. Samples from the WKY and WKY.LCrgn2 strains were amplified using a set of four biological replicates with three technical replicates per sample. ***P<0.001 statistically significantly different to WKY using a two way ANOVA to compare the overall timecourse with Bonferonni’s post-tests to compare individual time points.