Fig. 4: Oligomerization of CD163 is efficient for mediating internalization of Hb-Hp.

a SEC of CD163TriMut in the presence of 0 and 2.5 mM calcium. b SV-AUC of CD163TriMut in the presence of 2.5 mM calcium. The molecular weight of the peak in SV-AUC is ~139 kDa. c SV-AUC of CD163TriMut mixed with Hb-Hp1 at a molar ratio of 1:2. The frictional ratio is 1.51812. The molecular weights of the peaks in SV-AUC are ~138 and ~251 kDa. d Pull down analyses of Hb-Hp binding activity of recombinant CD163TriMut in the presence of 2.5 mM calcium, with each experiment repeated at least twice. e Western blot analyses of CD163 expression in HEK293T cells and HEK293T cells transfected with plasmids overexpressing full-length CD163. This experiment has been repeated twice. Uncropped blots in Supplementary Fig. 13. f Flow cytometry and imaging analyses of HEK293T cells binding to and internalizing Hb-Hp. g Flow cytometry and imaging analyses of HEK293T cells overexpressing CD163, showing binding to and internalizing Hb-Hp. h Flow cytometry and imaging analyses of HEK293T cells overexpressing CD163TriMut, showing binding to and internalizing Hb-Hp. Each flow cytometry experiment was repeated three times individually (marked with black dots on the bars). Data was plotted as the mean with SD. Statistical analysis is based on the two-tailed unpaired t-test (****, p  <  0.0001; **, p  =  0.0068). Source data are provided as a Source Data file.