Figure 4. Role of TRIM65 in dsRNA-induced activation of MDA5 signaling. (A–H) qPCR analysis of Ifn-β expression in Mda5−/− (A and G) or Trim65−/− (B–F and H) BMDMs stimulated with cytosolic LMW poly I:C (A and B), HMW poly I:C (A and C), cytosolic 3pRNA (D), extracellular LPS (E), poly I:C (F), and EMCV-RNA or VSV-RNA (G and H) for 6 h. (I) Secretion of IFN-β in Trim65−/− BMDMs stimulated with different doses of EMCV-RNA or VSV-RNA for 18 h. (J) Immunoblot analysis of IRF3 phosphorylation (p-IRF3) in WT or Trim65−/− BMDMs stimulated with cytosolic EMCV-RNA or VSV-RNA for 3 h. Data are from three independent experiments with biological duplicates in each (A–I; mean and SEM of n = 6) or are representative of three independent experiments (J). Statistics were analyzed via an unpaired Student’s t test: ***, P < 0.001.
Figure 4.

Role of TRIM65 in dsRNA-induced activation of MDA5 signaling. (A–H) qPCR analysis of Ifn-β expression in Mda5−/− (A and G) or Trim65−/− (B–F and H) BMDMs stimulated with cytosolic LMW poly I:C (A and B), HMW poly I:C (A and C), cytosolic 3pRNA (D), extracellular LPS (E), poly I:C (F), and EMCV-RNA or VSV-RNA (G and H) for 6 h. (I) Secretion of IFN-β in Trim65−/− BMDMs stimulated with different doses of EMCV-RNA or VSV-RNA for 18 h. (J) Immunoblot analysis of IRF3 phosphorylation (p-IRF3) in WT or Trim65−/− BMDMs stimulated with cytosolic EMCV-RNA or VSV-RNA for 3 h. Data are from three independent experiments with biological duplicates in each (A–I; mean and SEM of n = 6) or are representative of three independent experiments (J). Statistics were analyzed via an unpaired Student’s t test: ***, P < 0.001.

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