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. 2021 Jan 2;48(1):425–433. doi: 10.1007/s11033-020-06069-z

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© The Author(s) 2021

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/.

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Fig. 1 — Treatment of human rheumatoid fibroblast-like synoviocytes (RFLS) with polyinosinic-polycytidylic acid (poly I:C) induces the expression of interferon beta (IFN-β), melanoma differentiation-associated gene 5 (MDA5), and CXCL10 in a concentration-dependent manner. RFLS were cultured and treated with 0.08–50 mg/mL poly I:C. (a) RNA was extracted after 4-h incubation. cDNA was synthesized and IFN-β, MDA5, CXCL10, or GAPDH cDNA was amplified using quantitative real-time PCR. Expression of IFN-β, MDA5, and CXCL10 mRNA was normalized with GAPDH. Data for MDA5 was shown as fold increase vs. unstimulated cells. IFN-β and CXCL10 mRNA expression was undetectable in unstimulated cells, and the fold increase for IFN-β and CXCL10 is shown as arbitrary units. (b) Cells were lysed after 16-h incubation, and lysates were subjected to western blot to detect MDA5 and actin protein expression. (c) Conditioned medium was collected after 24-h incubation, and the concentration of CXCL10 protein in the medium was measured using an ELISA kit. The data in (a) and (c) represent means ± standard deviation (n = 3)