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. 2018 Mar 19;293(19):7195–7208. doi: 10.1074/jbc.RA118.002639

Figure 6.

Figure 6.

Levels of complement proteins, C3 and FB, and complement activation products, C3a, C5a, and Ba, in the supernatant of unstimulated and TNF-stimulated BMVECs and GMVECs. Supernatant from unstimulated BMVECs and GMVECs in T-75 flasks was collected and assayed by ELISA for complement components C3 and FB (A) and AP activation products C3a, C5a, and Ba (B and C). The same flasks were then stimulated for 48 h with 10 ng/ml TNF (24 h in complete media and 24 h in serum-free media), and the 24-h supernatant was again collected and assayed for the same proteins. A, n = 4 for BMVECs and n = 6 for GMVECs for C3, and n = 4 for both GMVECs and BMVECs for FB; FB levels in unstimulated BMVECs and GMVECs were below detection limits (bdl) of the assay. B, n = 6 for GMVECs and n = 4 for BMVECs for C3a experiments, n = 6 for both BMVECs and GMVECs for C5a experiments, and n = 4 for both BMVECs and GMVECs for Ba experiments; C3a levels in unstimulated and TNF-stimulated BMVECs were below detection limits of the assay. C, percentage of Ba generated from FB under TNF-stimulating conditions was calculated from Ba levels in B and FB levels in A; n = 4 for both BMVECs and GMVECs. *, p < 0.05; **, p < 0.001.