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. 2018 May 28;9:1191. doi: 10.3389/fimmu.2018.01191

Figure 2.

Figure 2

Immunoblotting for factor D (FD) in sera from 10- to 14-week-old MRL/lpr and C57BL/6 mice. Serum FD were purified by immunoprecipitation with a rabbit anti-mouse FD antibody, deglycosylated with N-glycosylase F, separated by SDS-PAGE, and blotted onto a PVDF membrane. Purified pro-FD (26.1 kDa) and FD (25.5 kDa) were detected with a horseradish peroxidase-conjugated rabbit anti-mouse FD antibody. Consistent with our previous report on C57BL/6 mice (8), serum FD in Masp1/3−/− MRL/lpr mice was in an inactive form (pro-FD) when tested for serum samples from three independent Masp1/3−/− MRL/lpr mice (#1–3). In contrast, serum FD in WT MRL/lpr mice was in an active form as well as in WT C57BL/6 mice.