Figure 1.

Analysis of cancer stem cell (CSC) properties in SCC4 and SCC9 cell lines. (A) Representative images of the gates used to sort the CD44^high and CD44^low fractions. (B) Cell morphology of CD44^high and CD44^low fractions, demonstrating the presence of holoclones in CD44^high cells. (C) Colony formation ability of CD44^high and CD44^low fractions. Graphs representing the (D) number of colonies and (E) number of spheres in CD44^high and CD44^low cells. (F) RT-qPCR for CD44, NANOG, CD133, POU5F1 (OCT4) and BMI1 transcripts. Fold change values represent the relative gene expression in CD44^high cells in relation to CD44^low cells. ^*P<0.05, ^**P<0.01 and ^***P<0.001 (t-tests with the Welch correction). Data for SCC9 cells was gathered as previously described by de Andrade et al (51). Microscope images: Magnification, ×100 (scale bars, 40 μm).