Supplementary Figure 1: IC261 inhibits a virus-induced type I interferon response. | Nature Immunology

Supplementary Figure 1: IC261 inhibits a virus-induced type I interferon response.

From: The kinase CK1ɛ controls the antiviral immune response by phosphorylating the signaling adaptor TRAF3

Supplementary Figure 1

(a) HEK293T cells were cultured in 384 wells and transiently transfected with 50 ng of the IFN-β promoter-Luc construct along with 5 ng TK-Luc construct for 12 hours. Various compounds were then added to culture media for 12 hours and luciferase activity was detected. Sample with a low TK activity (smaller than half of control) was considered to be cytotoxic and was ruled out. (b-i) Q-PCR analysis of DMSO- or IC261 (10 μM)-treated primary peritoneal macrophages stimulated by VSV (12h) and poly(I:C) (3h). (j-l) Q-PCR analysis of DMSO- or IC261 (10 μM)-treated primary peritoneal macrophages stimulated by VSV. (m) IB analysis of p52 in lysates of DMSO- or IC261-treated primary peritoneal macrophages infected with VSV. (n) Primary peritoneal macrophages cells were pretreated with DMSO or IC261 (10 μM) for 60 minutes, and then infected with VSV (MOI=0.01). The mRNA level of VSV was examined by Q-PCR analysis. (o) HeLa cells were pretreated with DMSO or IC261 (10 μM) for 60 minutes, and then infected with HSV (MOI=0.001). The mRNA level of HSV was examined by Q-PCR analysis. NS, not significant (P > 0.05); *P < 0.05, **P < 0.01 and ***P < 0.001 (unpaired t-test (a-h)). Data are from three independent experiments with biological duplicates in each (b-o; mean and s.e.m. of n = 3). Data are representative of three independent experiments (m).

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