Fig. 1 NDR2 facilitates RNA virus–induced IFN-β, IL-6, and TNF-α production via a kinase activity–independent mechanism in macrophages. NDR2 facilitates.

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Fig. 1 NDR2 facilitates RNA virus–induced IFN-β, IL-6, and TNF-α production via a kinase activity–independent mechanism in macrophages. NDR2 facilitates RNA virus–induced IFN-β, IL-6, and TNF-α production via a kinase activity–independent mechanism in macrophages. (A and B) IFN-β, IL-6, and TNF-α expression in NDR2f/f and Lysm+NDR2f/f PMs infected with VSV in the indicated times was detected by enzyme-linked immunosorbent assay (ELISA) (A) and real-time polymerase chain reaction (PCR) analysis (B). (C) Real-time PCR analysis of IFN-β, IL-6, and TNF-α transcripts in NDR2f/f and Lysm+NDR2f/f PMs treated with phosphate-buffered saline (PBS) or with the infection of H1N1, SeV, RSV, EMCV, HSV-1, or MHV68 for 8 hours. (D) Fluorescence-activated cell sorting (FACS) analysis of enhanced green fluorescent protein (eGFP) fluorescence intensity in NDR2f/f and Lysm+NDR2f/f PMs infected with VSV-eGFP. (E) VSV-eGFP titers by TCID50 assay in supernatants of NDR2f/f and Lysm+NDR2f/f PMs infected with VSV-eGFP for 6 hours. (F) NDR2f/f and Lysm+NDR2f/f PMs were transfected with empty vector (mock), NDR2, or its kinase-inactive mutants overexpressing plasmids for 36 hours and then infected with VSV, followed by real-time PCR analysis of IFN-β, IL-6, and TNF-α expression. AA, K282A/T442A. (G) FACS analysis of RAW264.7 cells stably overexpressing NDR2 or its kinase-inactive mutants infected with VSV-eGFP. Data are means ± SD and are representative of three independent experiments. Student’s t test was used for statistical calculation. ns, no significance. *P < 0.05, **P < 0.01, and ***P < 0.001. Zhiyong Liu et al. Sci Adv 2019;5:eaav0163 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).