Figure 5.

Brachyury silencing reduces the sphere-forming ability of ACCS-M GFP cells. ACCS cells and ACCS derivatives transfected with shRNAs (−, untreated control; con, control shRNA; Bra, Brachyury shRNA; Sox2, SOX2 shRNA) were cultured, and sphere-forming ability was quantified as described in the legend for Figure 1. Sphere diameters were measured (A), and spheres with a diameter >100 μm were counted. Sphere numbers were standardized as sphere number/10⁴ cells originally cultured (B) in each sphere period. Experiments were performed in triplicate, and the values were averaged. Bars indicate the standard deviation. Data significance was analyzed by Student’s t-test. *P < 0.05, **P < 0.01.