Fig. 3

Gene and protein expression change during the in vitro culturing of ASCs. a Heatmap represents the hierarchical clustering of the 2395 genes expressed differentially (− 2× > FC > 2× and FDR < 0.05) in the RNA-seq experiment. The investigated cell types are indicated at the bottom of the chart. Both groups consist of biological replicate pairs (a, b) with highly similar pattern, in contrast, vASC pair demonstrates strikingly different pattern compared to ASC.B6 pair. b Functional enrichment analysis done by IPA for the identification of biological functions and diseases that were most profoundly represented by the differentially expressed genes. Only carcinogenesis associated categories are presented, all of which are highly affected. Right-tailed Fisher’s exact test was used to calculate a p-value. False discovery rates (FDR) were generated based on the Benjamini-Hochberg corrected p-values. c Klf4 protein expression was analyzed with Western blotting in total cell lysates of vASCs harvested at P2 or P6 and ASC.B6 cells. The Klf4 signals were normalized to β-actin, and divided with the value of vASCs at P2. d Nestin protein expression was analyzed with Western blotting in total cell lysates of vASCs harvested at P3 or P7 and ASC.B6 cells. The Nestin signals were normalized to β-actin, and divided with the value of vASCs at P3. The pictures show a representative of at least 3 independent experiments