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. 2020 Jul 11;39:132. doi: 10.1186/s13046-020-01637-4

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — CPEB3 inhibits IL-6R/STAT3 signaling via direct binding to IL-6R mRNA in CRC cells (a) The effect of the TAMs on the JAK1, pJAK1, pSTAT3, and STAT3 in CRC cells (HCT116-Ctrl/CPEB3 and LoVo-shCtrl/shCPEB3) was analyzed by western blot analysis. (b) The effect of IL-6 on the IL-6ST, IL-6R, STAT3, and pSTAT3 in CRC cells (SW480-Ctrl/CPEB3 and HCT116-Ctrl/CPEB3) was analyzed by western blot analysis. (c) The effect of IL-6 on the IL-6ST, IL-6R, STAT3, and pSTAT3 in CRC cells (LoVo-shCtrl/shCPEB3 and RKO-shCtrl/shCPEB3) was analyzed by western blot analysis. (d) Schematic diagram of IL-6R-3′ -UTR reporter mRNA. (e) Luciferase assays were performed to detect the binding activity of CPEB3 and IL-6R. Relative fold-change in luciferase activity is shown; error bars, SEM. (f) RT-PCR of the RIP products confirmed the direct binding capacity of CPEB3 to the IL-6R-3′ -UTR in SW480 and LoVo cells. qRT-PCR of the RIP products further confirmed the direct binding capacity of CPEB3 to the IL-6-3′-UTR in SW480 and LoVo cells. Input, 5% of total lysate; error bars, SEM. (g) Cell Counting Kit-8 was used to quantify the number of LoVo/RKO-shCPEB3 cells, cultured TAMs supernatants, and TAMs supernatants treated with tocilizumab (5 ng/mL). (h) The invasion of LoVo/RKO-shCPEB3 and LoVo/RKO-shCPEB3 co-cultured TAMs treated with or without tocilizumab (5 ng/mL) was measured by a Transwell assay (200× magnification); error bars, SEM. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001