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. 2018 Aug 7;9:1813. doi: 10.3389/fimmu.2018.01813

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Copyright © 2018 Mohlin, Gros, Mercier, Gris and Blom.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Expression of C3 variants. (A) C3 variants, together with wild type (WT) and negative control (mock = empty vector), were transiently transfected into adherent human embryonic kidney (HEK) 293 cells. After four independent transfections, the C3 concentration was measured both in the conditioned cell media (supernatant) and the cell lysate using ELISA. The results are shown as mean + SD and the statistical significance of the differences between WT and mutants was determined using two-way ANOVA with Dunnett’s multiple comparisons test, *p < 0.05, ***p < 0.001. The C3 alterations identified only in patients are marked in gray. (B,C) After transient transfections, the same volume of each cell supernatant was also subjected to SDS-PAGE under both (B) non-reducing and (C) reducing conditions, followed by Western blotting. C3 was detected using a polyclonal anti-C3 antibody.