Figure 9.

TAMPs are differentially expressed and redistributed in response to in vivo TNF treatment. Mice were sacrificed at the indicated times after intraperitoneal injection of 5 μg of TNF. (A) Epithelial cells were isolated, and RNA was harvested for qRT-PCR analysis. MarvelD3 and tricellulin message increased 45 min after TNF treatment and remained elevated after 90 min (*p < 0.01), whereas occludin message content was not changed (p > 0.40). (B) SDS-PAGE immunoblot and densitometric analyses of jejunal epithelial lysates demonstrate increased marvelD3 and tricellulin protein after 90 min of TNF exposure (*p < 0.05). (C) Jejunum was frozen 120 min after TNF injection and immunostained for TAMPs (green), F-actin (red), and nuclei (blue). MarvelD3 and tricellulin protein expression increased and was enriched at the tight junction, lateral membrane, and cytoplasm. Bar, 5 μm. Data are representative of ≥3 experiments, all with similar results.