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Blood, Vol. 113, Issue 15, 3640-3648, April 9, 2009
C7 is expressed on endothelial cells as a trap for the assembling terminal complement complex and may exert anti-inflammatory function
Blood Bossi et al.
113: 3640
Supplemental materials for: Bossi et al
Files in this Data Supplement:
- Table S1. Peptides from C7 and vimentin observed in the tryptic digests of the affinity purified protein material (XLS, 876 KB)
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MM, calculated average molecular mass of the unmodified peptide sequence; z, charge state; ProtProb, ProteiProphet probability; Adj. PepProb, adjusted PeptideProphet probability; Init. PepProb., intial PeptideProphet probability; NTT, number of tryptic termini; #, oxidized methionine.
- Figure S1. Glycocalix removal did not reduce mC7 expression (JPG, 55.4 KB)
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HUVECs were treated with the mixture of the heparinase I, II and III (each at 3 × 10−2 Units/ml) for 2 h at 37°C to remove cell surface glycosaminoglycans. (A) The presence of residual cell-bound C7 and CD31, used as control of an integral membrane protein, was revealed by a fluorescence reader Infinite200 (TECAN) using FITC-labeled antibodies.The removal of glycocalix was checked by the failure of FITC-Ulex europaeus agglutinin I (UEA-I) to bind to treated cells. Values are mean ± SD of triplicate determinations of 3 separate experiment. *P <.05. (B). FACS analysis of heparinase-treated (black line) and untreated cells (dotted line) for the expression of mC7. One of 3 different experiments with similar results is shown.
- Figure S2. SDS-resistant polyC9 in mTCC (JPG, 96.6 KB)
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Native C9 (track 1) or EC lysate collected after the assembly of mTCC were analyzed by Western blot with polyclonal anti-C9 (1:100) followed by AP-conjugated anti-goat antibodies. Data are representative of three independent experiments.
- Figure S3. mC7 expression is increased by pro-inflammatory cytokines (JPG, 44.2 KB)
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HUVECs were stimulated with IL-1α (10 U/ml) , IFN-γ (100 U/ml), or TNF-α (100 ng/ml) for 48 h and the presence of mC7 was evaluated by ELISA. Values are mean ± SD of triplicate determinations of 3 separate experiment. *P <.05 versus basal value.
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