Figure 2

Figure 2

Cellular internalization of mAb and mAb-AMFA in cells and M6PR-dependency. After conjugation with AlexaFluor647^® dye, 0.75 µg.mL^-1 IFX or IFX-AMFA1 were incubated with Jurkat or macrophage-like cells in the presence of 0.188 µg.mL^-1 TNF-α. Internalization of mAb quantified by fluorescence measurement by flow cytometry at different time points (A). Involvement of M6PR in IFX-AMFA1 endocytosis. An AMFA excess was added to saturate M6PR and the fluorescence was studied by flow cytometry in Jurkat cells (B) or by confocal microscopy in macrophage-like cells (C). Quantification of confocal microscopy was performed on an average of 6-7 images consisting each of 100 cells per condition (D). Results are expressed as a percentage ± SD of mAb internalization relatively to IFX (A, B) or as mean of intensity ± SD (D). Student’s T-test: ** p value < 0.01.