Fig. 4

RT-qPCR analysis of stemness genes. When HT-29 and Caco2 cells were exposed to 5-FU, the expression of genes that regulate stemness was found to be higher than in the parental cell population. A) Compared to the spheroids, the isolated Caco2 subpopulation exposed to 5-FU showed a significant increase in C-MYC and OCT4 expression (p-value < 0.0001). B) With the exception of NANOG, the expression levels of stemness genes in the isolated HT-29 subpopulation cells exposed to 5-FU did not differ significantly from those in the spheroids. Interestingly, spheroid cells showed a more marked upregulation of NANOG (p-value < 0.0001) in comparison to the isolated HT-29 subpopulation cells exposed to 5-FU. The baseline gene expression in parental cells is represented by a dotted line. Data are presented as mean ± SD from three independent experiments as **** = p-value < 0.0001