Figure 6.

ARTN enhanced stem cell-like properties in DLD1 and HCT116 cells. (A) Serial colonosphere formation assay with DLD1 with either stable forced (left) or depleted (right) expression of endogenous ARTN. The colonospheres were passaged successively twice (P1-P3). For each passage of the colonospheres, the initial singular 100 cells were seeded in each well with triplicates and cultured for 7 days before re-seeding. The number of colonospheres (≥100µm in diameter) in each well was counted and presented as the average of the triplicate. The images of the P1 colonospheres were captured under a microscope. Scale bar, 100 µm. (B) Western blotting analysis for the expression of ARTN in the whole cell lysate of monolayer culture or colonospheres of DLD1 cells. (C), ALDEFLUOR (ALDH1 activity) assay on DLD1 and HCT116 cells with stable forced expression of ARTN (left) or depletion of endogenous ARTN (right). *p < 0.05, **p < 0.01.