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Fig. 1. In HUVECs, the ectodomain-shedding of TNFRs involves metalloprotease activity and is increased by proteasome inhibition. Ro 32-7315 (5 µM) was added or not to HUVECs 15 minutes before the cells were incubated for 4 hours with MG-262 (600 nM) or lactacystin (40 mM) as indicated. (A) TNFR1 and TNFR2 that had accumulated in the culture media was measured by ELISA. (B) Cell surface expression of TNFR1 and TNFR2 was analyzed by flow cytometry. Cont, control; Lac, lactacystin; Ro, Ro 32-7315. Flowcytometry experiments were performed in duplicate and repeated three times. Significance of difference between treated and control cells: *, P<0.05 and **, P<0.001.





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