Fig. 3

Validation of RNA-sequencing results. a Comparison of DE genes using RNA-seq and qPCR results using a log2FC cut-off ± 2. Significance threshold for RNA-seq p-adj values < 0.01 as routinely used. Significance threshold for qPCR p-value < 0.05 as routinely used. Red shaded boxes indicate no significant difference. Yellow shaded boxes represent a significant p-adj value; however, they do not meet the log2FC cut-off. Green shaded boxes indicate there is a significant difference based on both p-adj/p value and log2FC. NA values present typically arise due to count outliers as detected by Cook’s distance. b Immunocytochemistry comparison of the corresponding proteins for 12 significantly DE genes. Images represent longitudinal sections of 3D adult, fetal and ESC seeded constructs following 14 days of culture (n = 3 for each cell type). RNA-sequencing log2FC are listed in the adjoining tables with green text indicating significance based on p-adj value of < 0.01 and red text indicating no significant difference. Scale bar = 40 μm. DAPI staining of the nuclei is shown in blue