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. 2017 Feb;214(2):459–473. doi: 10.1084/jem.20160592

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© 2017 Lang et al.

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Figure 9. — EMCV-induced MDA5 oligomerization depends on TRIM65. (A and B) Confocal microscopy analysis (A) or qualification of aggregates (B) in HeLa cells stably expressing shRNA against TRIM65 that were transfected with mCherry-MDA5 for 24 h and then stimulated with EMCV-RNA for 3 h. Bar, 20 µm. (C) Immunoblot analysis of MDA5 by SDD-AGE or SDS-PAGE assay in HeLa cells stably expressing shRNA against TRIM65 that were transfected with Flag-MDA5 for 24 h and then stimulated with EMCV-RNA for 3 h. (D) Immunoblot analysis of MDA5 by SDD-AGE or SDS-PAGE assay in WT or Trim65^−/− BMDMs that were primed with LPS for 12 h and then transfected with EMCV-RNA for 3 h. Data are from two independent experiments with biological duplicates in each (mean and SEM of n = 4; B) or are representative of two or three independent experiments (A, C, and D). Statistics were analyzed via an unpaired Student’s t test: ***, P < 0.001.