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. Author manuscript; available in PMC: 2021 Aug 14.
Published in final edited form as: Cell Rep. 2021 Jun 22;35(12):109272. doi: 10.1016/j.celrep.2021.109272

Figure 3. Riok3 deficiency selectively upregulates the production of RIG-I/MDA5 activation-induced anti-viral cytokines in PMs and MEFs.

Figure 3.

(A–C) qRT-PCR analysis of IFN-β (A), TNF-α (B), and IL-6 (C) in Riok3+/+ and Riok3−/− peritoneal macrophages transfected with 5 μg/mL short-length poly(I:C) for indicated hours.

(D–F) qRT-PCR analysis of IFN-β (D), TNF-α (E), and IL-6 (F) in Riok3+/+ and Riok3−/− peritoneal macrophages transfected with 5 μg/mL long-length poly(I:C) for indicated hours.

(G–I) ELISA of IFN-β (G), TNF-α (H), and IL-6 (I) in supernatants of Riok3+/+ and Riok3−/− peritoneal macrophages transfected for 12 h with 5 μg/mL short-length poly(I:C), 5 μg/mL long-length poly(I:C), 1 μg/ml poly(dA:dT), 5 μg/mL 3′3′-cGAMP, and 5 μg/mL CpG DNA, respectively.

(J) Immunoblot analysis of phosphorylated or total proteins in lysates of Riok3+/+ and Riok3−/− peritoneal macrophages transfected for indicated times with two forms of poly(I:C). Phosphorylation of TBK1 Ser172, IRF3 Ser396, and p65 Ser536 was detected.

(K–M) ELISA of IFN-β (K), TNF-α (L), and IL-6 (M) in supernatants of Riok3-deficient and control MEFs transfected with 5 μg/mL long-length poly(I:C) for 12 h. 2 μg/mL shRiok3 or control plasmids was transfected into MEFs for 40 h to knock down Riok3 expression before poly(I:C) transfection.

(N) qRT-PCR analysis of anti-viral ISGs in Riok3 knockdown or control MEFs transfected with 5 μg/mL long-length poly(I:C) for 6 h.

Data are shown as mean ± SEM. Results are representative of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001.