Fig. 8

RT-qPCR validation of gene expression changes detected using mRNA-seq. Confirmation of differentially expressed genes from NGS results using quantitative real-time PCR. Significant changes in gene expression of both a pro- and anti-inflammatory cytokines as well as other b effector molecules of the immune response confirmed the findings from NGS. Results are colour-coded according to comparison and levels of expression of each gene of interest was normalised to expression levels of PPIA; D7PP between SCE and HC cows (black), D21PP between SCE and HC (blue), between D7 and D21PP in SCE (red) and between D7 and D21PP in HC (green). Between and within group comparisons are separated by a dotted line. *P < 0.05; **P < 0.01. n = 5–8 samples per time point, bars represent mean ± SEM