Fig. 9: The impact of paternal per- and polyfluoroalkyl substances (PFAS) exposure on embryonic gene expression in 4-cell embryos.

Volcano plots illustrate the gene expression profiles in 4-cell embryos fertilized by control spermatozoa or sperm from male mice exposed to A low and B high PFAS. Genes with p ≤ 0.05 and fold-changes ±1.5 were considered to have differential gene expression (DEGs) and are denoted by either pink or purple dots to identify those as being significantly up- and down-regulated. Alternatively, green dots were used to denote those DEGs with expression that changed consistently across both exposure regimens. Interrogation by ingenuity pathway analysis (IPA) software was used to identify the diseases predicted to be affected by embryonic DEGs (p ≤ 0.05) in the C low and D high PFAS groups. The displayed heatmaps are based on a p < 0.05 and a Z-score scale where a score of ≥2 is considered “activated” and a score ≤ −2 is considered “inhibited”. Tables showing the genes targeted by altered sperm microRNAs (miRNAs) in the E low and F high PFAS groups. Created in BioRender. Nixon, B. (2025) https://BioRender.com/eq3tqoz (Agreement number: QT28OPEW0P).