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Fig. 3 | Cell Communication and Signaling

Fig. 3

From: TNF-α promotes human antibody-mediated complement-dependent cytotoxicity of porcine endothelial cells through downregulating P38-mediated Occludin expression

Fig. 3

Expression of complement-regulatory proteins on PIECs was increased, and the extent of human IgG and IgM binding to PIECs, and C3 or C5b-9 deposition on PIECs was not affected by rhTNF-α. a PIECs were treated with rhTNF-α (20 ng/ml) for 0, 2, 6 or 12 h. The mRNA levels of CD46, CD55, or CD59 were measured by RT-PCR. b-c PIECs were treated with rhTNF-α (20 ng/ml), or medium as a NC for 48 h, and then incubated with human serum for 30 min. The expression of CD46 (b), CD55 (b), C3 deposition (C3 and C3c) (c) or C5b-9 deposition (c) was assessed by flow cytometry. The quantitation data were presented by geometric mean fluorescence intensity (Gmean). d-e PIECs were treated with rhTNF-α (20 ng/ml), or medium as a NC for 48 h, and then incubated with heat-inactivated human serum (HIHS) or heat-inactivatd human serum minus IgG, IgA and IgM (HIHS[−]) for 30 min. Human IgM or IgG binding to PIECs was measured by flow cytometry (d). The extent of human IgM or IgG binding to PIECs was evaluated by Gmean (e). Data are representative of at least three independent experiments (mean ± SEM). *p < 0.05, **p < 0.01 by Student’s t test. N.S. = no significance

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