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. 2021 Apr 21;476(9):3241–3252. doi: 10.1007/s11010-021-04157-7

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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. 5 — MDA5 and RIG-I regulate pro-inflammatory cytokines in human CF after intracellular PolyIC stimulation. (a-b) Knockdown of MDA5 and RIG-I with siRNA in human CF stimulated for 2, 4, and 8 h with 1 ng/ml of intracellularly transfected PolyIC. SiMock was used as a control. Transcription of IL-1β, IL-6, IL-8, but not of TNF-α, after stimulation with cytosolic PolyIC, was MDA5- and RIG-I-dependent (a). MDA5 and RIG-I knock-down efficiency was also determined by real-time RT-PCR (b). (c) Knockdown of TLR3, MDA5, and RIG-I with siRNA in human CF stimulated for 24 h with 1 ng/ml of intracellularly transfected PolyIC. SiMock was used as a control. IL-6 and IL-8, but not IL-1β and TNF-α, all measured at the protein level in cell-free supernatants, showed reduced levels after siRNA-induced knockdown. n.d., not detected. Means ± s.d. and values measured from one out of two to three independent experiments performed in duplicates are shown. Two-way ANOVA with Bonferroni post hoc testing, ***p < 0.001 for siMock vs. siMDA5, ⁺⁺⁺p < 0.001 for siMock vs. siRIG-I (a); ***p < 0.001 for siMock vs. siMDA5 (left panel) or siMock vs. siRIG-I (right panel) (b); *p < 0.05, **p < 0.01 for siMock vs. siMDA5 and siMock vs. siRIG-I, ^#p < 0.05 for siTLR3 vs. siMDA5 and siTLR3 and siRIG-I (c)