Fig 3.

TGEV infection induces inflammatory responses by RIG-I in apical-out porcine intestinal organoids. (A) Transcription levels of RIG-I, MDA5, TLR3, TLR7, and TLR8 in apical-out organoids at the indicated times post-TGEV infection were evaluated by RT-qPCR. (B) RIG-I expression in apical-out organoids at the indicated times post-TGEV infection was detected by Western blotting and calculated with ImageJ. (C) Apical-out organoids were treated with 500 µM or 1 mM Cyclo for 48 h, then RIG-I expression was measured by Western blotting and calculated with ImageJ. (D–F) Apical-out organoids were infected with TGEV followed by Cyclo treatment (1 mM) for 48 h. Subsequently, mRNA levels of TNF-α, IL-8, IL-6, and IL-1β (D), TGEV viral load (E), and mRNA levels of IFN-α, IFN-β, and IFN-λ1 (F) were determined by RT-qPCR. Results are presented as mean ± SD of data from three independent experiments *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001, determined by two-tailed Student’s t test.