Fig. 1.

ISO inhibition of BC stem-like properties through inhibiting CD44 was also important for changes in invasivity. a, b T24T cells were treated with ISO at indicated concentrations for 7 days to determine impact on sphere formation ability. The number of spheroid-forming cells in each image was counted as described in “Materials and methods”. Bars shown are mean ± SD from three independent experiments. *Significant difference relative to controls (p < 0.05). c, d T24T cells were treated with ISO at indicated concentrations or for indicated times, and then evaluated for stemness marker protein expression using Western blots. e Over-expression of CD44 constructs were stably transfected into T24T cells; the transfectants were then treated with 20 μM ISO for 12 h, and then underwent Western Blot analyses to determine CD44 expression levels. f Over-expression of CD44 could not reverse ISO inhibition of sphere formation. *Significant difference between indicted cells treated with or without 20 μM ISO (p < 0.05). g, h Invasivity of T24T(vector) and T24T(CD44) cells treated with or without 20 µM ISO for 24 h was evaluated using a BD BioCoat TM Matrigel TM Invasion Chamber. After incubation for 24 h, cells were fixed and stained as described in “Materials and methods”. *Significant difference (p < 0.05)