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. 2020 Jul 14;161(1):66–79. doi: 10.1111/imm.13229

Figure 1.

Figure 1

IgG anti‐αGal activates the classical pathway on pig red blood cells (pRBCs). Complement deposition on fixed pRBCs measured by flow cytometry using detection antibodies against C4c (which recognize C4b and iC4b fragments) (a) or C3c (which recognize C3b and iC3b fragments) (b). The pRBCs were incubated for 2 hr at 37° in 1% hypogammaglobulinaemia human serum (HHS) as the complement source supplemented with IgG anti‐αGal at 20 mg/l and inhibitors as indicated. Inhibitors were C1qNb75 or KRA152 nanobody. C1qNb75 targets the globular heads of C1q, thereby blocking C1q docking to immunoglobulin and inhibiting the classical pathway of complement. KRA152 possess irrelevant specificity and was included as a control. MFI, median fluorescence intensity. The columns represent the mean of four separate experiments (each represented by a dot) and error bars are 95% confidence intervals.