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. Author manuscript; available in PMC: 2019 Jul 28.
Published in final edited form as: Nat Med. 2019 Jan 28;25(3):496–506. doi: 10.1038/s41591-018-0336-8

Extended Figure 3. ApoE does not inhibit cleavage of C2 or C4 by C1s.

Extended Figure 3

(a) C1q binds immobilized malondialdehyde-modified LDL (MDA-LDL) and oxLDL but not native LDL or gelatin. (b) ApoE isoforms in NHS were added to MDA-LDL-coated microtiter plates and C4b deposition was determined by specific antisera. (c,d) IgM, MDA-LDL, and Aβ fibrils but not soluble Aβ activate complement and cause C3b deposition. BSA, gelatin as negative controls; (e, f) ApoE3 was incubated with either (e) C2 or (f) C4 in the presence of C1s. C2 and C4 were cleaved to their active forms C2a (α′30) and C4b (α′83) via C1s as revealed by the cleavage products in Western blot analyses; (g) ApoE3 has no cofactor activity for factor I in the cleavage of C4b to inactive iC4b. ApoE3 was incubated together with factor I, C4BP and C4b, and cleavage products were detected by Western blot analysis as indicated (α′25 and α′13). Full scanned blot images in e,f,g are available from source data figures. Data in a-d represent means ± SEM of three independent experiments. Two-tailed Student's t-test. Data in e,f,g are representative from 3 independent experiments.