Figure 4.

Determining the effect of SIX1 on sphere formation and self-renewal ability. (A) These cells were passaged 2 times to determine the effect of SIX1 on the self-renewal capacity following the sphere formation assay. It was subsequently observed that the shSIX1-SNU398 cells were unable to self-renew. (B) RT-qPCR was used to analyze the expression levels of SIX1, ZEB2, VIM, KLF4, and THY1 genes in sphere-forming cells. (C) CD90 cell population in shSIX1-SNU398 and control-SNU398 cells was analyzed using flow cytometry. The CD90⁺ subpopulation was decreased in shSIX1-SNU398 cells compared with the control-SNU398 cells. The differences between groups were analyzed using a Student’s t-test. SIX1, SIX1, Sine oculis homeoprotein 1. ^** P < .01, ^*** P < .0001. Magnification, ×5; scale bars, 200-μm.