FIGURE 3.

Tofacitinib rescues the permeability increase induced by IFN-γ in human colonic organoids. A, Representative series of optical slices through a human colonoid bathed in growth media containing 0.1 mg/mL FD4. Scale bar = 100 μm. B, Following a rescue protocol, human colonoids cultured for 12 days were treated with IFN-γ (50 ng/mL, administered in the bathing media). Tofacitinib (16.7 μM, administered in the bathing media) was added 6 hours, 24 hours, and 48 hours post-IFN-γ treatment. FD4 flux across colonoid epithelium normalized to surface area of colonoids was measured 72 hours after IFN-γ administration. (*** P < 0.001, ## P < 0.01, n = 4). C, Colonoids were harvested, lysed, and subjected to Western blotting for ZO-1 and occludin. Representative blots and densitometric analysis for ZO-1 and occludin are shown.