Quantification of cell surface molecules.

CD40–B cells were generated by means of tCD40L or sCD40L in the presence of IL-4 and CsA and were coincubated with or without INF-γ (added for 4 days before analysis). DCs were generated by culture of monocytes with GM-CSF and IL-4 and were matured with TNF-α or CD40L/INF-γ. (A) Surface expression of CD58, CD80, MHC class I, and MHC class II was measured by flow cytometry and normalized by means of the Qifikit to obtain the ABSs per cell. (B) The number of ABSs per cell surface area was calculated to normalize for different sizes, cell types, or culture conditions. Medians (lines), ranges (bars), and 95th percentiles (boxes) of 3 to 9 experiments for each culture condition and cell surface molecule are shown.