FIGURE 4.

Aldefluor-positive cells show increased tumorsphere formation and GnT-V expression. A, Aldefluor-positive and -negative were sorted and “stemness”-associated genes were detected by qRT-PCR from LS180 cells (A) and HT-29 cells (B), respectively. ALDH, aldehyde dehydrogenase. C, sorted Aldefluor-positive and -negative HT-29 cells (4000 cells/well) were grown in stem cell culture media in suspension for 7–10 days, and the number of tumorspheres was counted in 5–6 random fields. Bar, 50 μm. D, GnT-V mRNA levels (left) and N-linked β(1,6) branching (right) were determined by qRT-PCR and L-PHA staining, respectively, in both Aldefluor-positive and -negative LS180 cells. For each transcript, the values was normalized to control (GAPDH) and expressed as the mean ± S.D. of three independent experiments. Images are representative of two independent experiments. Bar, 50 μm.