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. 2020 Dec 5;11(12):1035. doi: 10.1038/s41419-020-03241-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Schematic for IL-6 conditioning of MB cells. The chemosensitive Med8A-S cells were cultured for 4 weeks in media supplemented with 2 ng/mL IL-6. Following the conditioning stint, the cells were cultured a further 2 weeks without exogenous IL-6 to yield Med8A-IL6+ variant used for subsequent assays. b Western blot analysis of Med8A-S and Med8A-S-IL6+ cells for pY705 and total STAT3 levels under basal conditions, or transiently stimulated with 5 ng/mL IL-6 for 10 mins. c Analysis of Med8A-S, Med8A-R, and Med8A-S-IL6+ cells using flow cytometry for IL6R protein expression (Left panel), and QPCR for IL6R mRNA expression (Right panel). Error bars represent mean ± SD (n = 3); **p < 0.01, ***<0.001, two tailed unpaired t-test. Cell viability assay to assess the sensitivity of (d) Med8A-S and Med8A-S-IL6+ cells, (e) Med8A-R, Med8A-R-IL6+, IL6R^−/−, and IL6R^−/−-IL6+ cells, and, (f) Med8A-R, STAT3^−/− and STAT3^−/−-IL6+ cells to vincristine. As plotted is the mean ± SD of an experiment performed in triplicates, representative of at least 3 independent experiments. ***p < 0.001, two-way ANOVA with Tukey’s multiple comparison test.