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. 2021 Oct 23;12(11):988. doi: 10.1038/s41419-021-04268-5

Fig. 3. IL-6R blockade with Tocilizumab suppresses Cisplatin-induced stemness in vitro.

Fig. 3

A Western blot analyses of Bmi-1, IL-6R, gp130, pSTAT3, and STAT3 in UM-SCC-1, UM-SCC-22A, and UM-SCC-22B cells treated with vehicle, rhIL-6 (20 ng/ml), Cisplatin (1 µM), and/or Tocilizumab (0.1 µM). B Western blot analysis of UM-SCC-1, UM-SCC-22A, and UM-SCC-22B cells treated with rhIL-6 (20 ng/ml) combined with vehicle, Cisplatin (1 µM), and/or Tocilizumab (0.1 µM) for 24 h, followed by additional rhIL-6 (20 ng/ml) for 24 h. C Western blottings to evaluate the effect of 1 µM Cisplatin and/or 0.1 µM Tocilizumab on UM-SCC-22A cells sorted for ALDH/CD44. D Stem cell marker protein array analysis of UM-SCC-1, UM-SCC-22A, and UM-SCC-22B cells treated with Cisplatin (1 µM) and Tocilizumab (0.1 µM), as compared to vehicle controls. E Western blot analysis of stem cell markers (Oct4 and Nanog) in UM-SCC-1 and UM-SCC-22B cells treated with rhIL-6 (20 ng/ml), Cisplatin (1 µM), and/or Tocilizumab (0.1 µM). F Representative images of immunofluorescence staining (Bmi-1 and ALDH1) of UM-SCC-22A cells plated in chamber slides and treated with Cisplatin (0–1 µM) and/or Tocilizumab (0–0.1 µM). G Graph quantifying mean cellular fluorescence of ALDH1 expression normalized to DAPI in UM-SCC-22A cells treated with Cisplatin (1 µm) and/or Tocilizumab (0.1 µM). H Graph quantifying mean cellular fluorescence of Bmi-1 normalized to DAPI stain in UM-SCC-22A cells treated with Cisplatin (1 µM) and/or Tocilizumab (0.1 µM).