Fig. 2

Division of DEGs into clusters based on expression patterns among treatment groups; validation of RNAseq results. a Heatmap showing relative expression of transcripts based on normalized read counts for all DEGs with log₂ fold changes > 1 or < − 1 and padj < 0.05 among PEG and NC groups (2180 DEGs total). Red = high expression; blue=low expression. The heatmap is split into 4 distinct clusters of expression patterns among treatment groups, based on k-means clustering: cluster 1 (highest expression in PEG), cluster 2 (high expression in both PEG and Unop), cluster 3 (highest expression in Unop), and cluster 4 (highest expression in NC). b Heatmaps showing the top 15 DEGs in each cluster, ranked by padj (PEG vs NC comparison). c Comparison of log₂ fold changes between RNAseq and RT-qPCR for 10 upregulated and 10 downregulated transcripts of interest (comparing PEG vs NC) that are represented in clusters 1, 2, or 4. Data represents the mean ± SEM (n = 3 animals per treatment group). d Correlation of RNAseq and RT-qPCR results from (c), using linear regression and Pearson correlation analyses. Each point indicates a specific transcript