Figure 3
Figure 3. Antibodies to DEC205 and MR are accumulated more efficiently by DCs than anti-CD40. (A) BDCA1+ DCs or (B) MoDCs were continuously incubated with 1 μg/mL fluorescently labeled anti-MR, anti-CD40, or anti-DEC205 for the indicated times, washed, counterstained, and fixed (top panels). Middle and bottom panels show data from the top panel, together with matched samples treated with leupeptin and NH4Cl (inh) during the continuous incubation. MuIso is the isotype for anti-CD40, and huIso is the isotype for anti-MR and anti-DEC205. Graphs depict the mean ± SEM normalized MFI from ≥ 3 independent donors. Normalized MFI indicates amount of accumulation; MFI was calculated by removing the contribution of surface fluorescence (4°C control) and normalizing the MFI for the number of fluorophores per antibody. Differences in antibody accumulation in the absence or presence of inhibitors were assessed with the paired t test, and statistically significant differences are depicted, *P < .05.

Antibodies to DEC205 and MR are accumulated more efficiently by DCs than anti-CD40. (A) BDCA1+ DCs or (B) MoDCs were continuously incubated with 1 μg/mL fluorescently labeled anti-MR, anti-CD40, or anti-DEC205 for the indicated times, washed, counterstained, and fixed (top panels). Middle and bottom panels show data from the top panel, together with matched samples treated with leupeptin and NH4Cl (inh) during the continuous incubation. MuIso is the isotype for anti-CD40, and huIso is the isotype for anti-MR and anti-DEC205. Graphs depict the mean ± SEM normalized MFI from ≥ 3 independent donors. Normalized MFI indicates amount of accumulation; MFI was calculated by removing the contribution of surface fluorescence (4°C control) and normalizing the MFI for the number of fluorophores per antibody. Differences in antibody accumulation in the absence or presence of inhibitors were assessed with the paired t test, and statistically significant differences are depicted, *P < .05.

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