Figure 4.

Blockade of IL6R signaling inhibits the invasion, migration and tumorigenicity of bladder CSCs. (A) A total of 2×10⁴ cells were placed into the upper chamber, and for each experiment, conditioned medium from one of the different groups was added to the bottom chamber as a chemoattractant. After a 48-hour incubation at 37 °C, the cells that invaded through the membrane or Matrigel were photographed and counted after staining with 0.1% crystal violet for 15 min. Representative images of the invasion and migration assays in the spheres treated with PBS, human recombinant IL6 (50 ng/mL) or human recombinant IL6 plus tocilizumab (5 µg/mL). (B) Representative subcutaneous tumors from the mice after 6 weeks of transplantation are shown, as evaluated by bioluminescent imaging. (C) Tumor volume was measured once a week when tumors were palpable. Representative results from three independent experiments are shown, and all the values are expressed as the means ± SD, *P<0.05, **P<0.01. CSC, cancer stem cell; IL6, interleukin 6; IL6R, IL6 receptor.