Fig. 2

MHC class II gene expression in colonoids after treatment with the three major proinflammatory cytokines. (A) A schematic representation of the flow of cytokine treatments. The colonoids were initially cultured for 2 days in EM, followed by changing the medium from EM to DM. On day 4, colonoids were treated with 30 ng/mL of recombinant canine IFN-γ, TNF-α, or IL-1β in either EM or DM for 24 h. This schematic was created using BioRender.com. (B) The representative image of the canine colonoids at 24 h after treatment with 30 ng/mL of IFN-γ, IL-1β and TNF-α in EM and DM. Scale bar = 50 μm. (C) DLA-DRA expression was quantified using RT-qPCR 24 h after cytokine treatment (30 ng/mL IFN-γ, IL-1β, or TNF-α) in EM or DM. Three biological replicates were used for this analysis, with each cytokine treatment performed in triplicate. Error bars represent the SEM. **p < 0.01