Journal: The American Journal of Pathology
Article Title: The Critical Role of TAK1 in Accentuated Epithelial to Mesenchymal Transition in Obliterative Bronchiolitis after Lung Transplantation
doi: 10.1016/j.ajpath.2012.02.022
Figure Lengend Snippet: TNF-α accentuates TGF-β1–driven EMT via TNFR1. A: Flow cytometry analysis demonstrates that PBECs express TNFR1 on their cell surface, but express little to no TNFR2. B: Stimulation of PBECs (n = 6) with soluble TNF-α (TNFsol), membrane-bound TNF-α (TNFcys), TNFR1-specific mutant (Cys-TNF32W/86T) membrane-bound TNF (TNFcysR1), or TNFR2-specific mutant (Cys-TNF143N/145R) membrane-bound TNF (TNFcysR2) (all 20 ng/mL) for 72 hours alone has no effect on the expression of E-cadherin, vimentin, or fibronectin. Stimulation, however, with TNFsol, TNFcys, or TNFcysR1, but not TNFcysR2, increases pro-MMP-9 secretion. TGF-β1 (10 ng/mL) down-regulates the expression of E-cadherin, increases the expression of vimentin and fibronectin, and increases the secretion of pro-MMP-9. Co-stimulation of the cells with TGF-β1 and TNFsol, TNFcys, or TNFcysR1, but not TNFcysR2, accentuates the changes in EMT marker expression compared to TGF-β1 alone. C: Co-stimulation of PBECs with TGF-β1 and TNFsol, TNFcys, or TNFcysR1, but not TNFcysR2, accentuates changes in cell morphology and EMT marker expression compared to TGF-β1 alone. FL1-H, height of fluorescence intensity; FSC-H, height of forward scatter; SSC-H, height of side scatter.
Article Snippet: Antibodies TNFR1 (HM2020; Hycult, Uden, The Netherlands) and TNFR2 (HM2007; Hycult) 21 primary antibodies were used to detect receptor expression by flow cytometry.
Techniques: Flow Cytometry, Mutagenesis, Expressing, Marker, Fluorescence