Fig. 7

Orthogonal validation of genes differentially regulated following acrylamide exposure in the mouse seminal vesicles. RNA-sequencing data was validated using quantitative PCR on a selection of 8 genes identified in the mouse seminal vesicle transcriptome (n = 4–6 individual mice per treatment group). Candidate genes were selected based on those differentially regulated and associated with key canonical pathways and upstream regulators, including A C-C motif chemokine (Ccl8), B Complement component 4b (C4b), C Collagen, type V, alpha 1 (Col5a1), D Cathepsin E (Ctse), E Early growth response 1 (Egr1), F Heart development protein with EGF-like domains 1 (Heg1) G Interferon alpha-inducible protein 27 (Ifi27), and H Ribosomal protein, large, P0 (Rplp0). Quantitative PCR data was processed using the delta C(t) method using Peptidyl-prolyl cis-trans isomerase A (Ppia) as the reference gene and then normalised to the control group. Data are presented as a column graphs with mean ± SEM values with corresponding transcriptomics data overlaid onto these data using a line graph. qPCR data were analysed by non-parametric Mann Whitney U test. * p ≤ 0.05 indicates statistical significance when comparing acrylamide to control. I Table of qPCR validated genes showing direct comparison between RNA-sequencing fold-changes and qPCR fold-changes. All graphical components were generated using Prism (version 9.0.0)