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. 2019 Apr;8(2):557–566. doi: 10.21037/tcr.2019.03.16

Figure 3.

Figure 3

The IL6 signaling pathway is activated in bladder CSCs. (A,B) Western blots were performed to detect p-STAT3 and STAT3 expression in the adherent cells (A) and spheres (B) treated with PBS, human recombinant IL-6 (10 ng/mL) or human recombinant IL6 plus tocilizumab (5 µg/mL). (C) Primary and secondary spheroid assays were performed in the spheres treated with PBS, human recombinant IL6 (50 ng/mL) or human recombinant IL6 plus tocilizumab (5 µg/mL). The total numbers of spheroids from 5,000 cells were counted. (D) The expression of stem-like associated genes (CD133, CD44, ABCG2, Oct-4 and Nanog) was assessed by qRT-PCR in the RT4 or J82 cells treated with PBS, human recombinant IL6 or human recombinant IL6 (50 ng/mL) plus tocilizumab (5 µg/mL). (E,F) The percentage of CD44 or CD133 in the RT4 or J82 cells treated with PBS, human recombinant IL6 or human recombinant IL6 (50 ng/mL) plus tocilizumab (5 µg/mL), as assessed by flow cytometry. Representative results from three independent experiments are shown, and all the values are expressed as the means ± SD, *P<0.05, **P<0.01 and ***P<0.001. CSC, cancer stem cell; qRT-PCR, real-time quantitative reverse transcription polymerase chain reaction; IL6, interleukin 6.