Latex beads, carboxylate-modified polystyrene, and fluorescent red (L3030, diameter 2 μm; Sigma) were coated for 1 h at 37°C with mouse IgG (Jackson ImmunoResearch). After rinsing and reconstituting in RPMI 1640 complete medium, beads were added to BMDCs at a ratio of 10:1 (beads:BMDCs) and incubated with BMDCs for indicated time periods at 37°C. BMDCs were collected, washed in cold PBS to remove nonadherent beads, and fixed in 4% (w/v) polyoxymethylene. Cells were subjected to flow cytometry analysis.
Endocytosis and Phagocytosis Assays
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Corresponding Organization : Peking University
Other organizations : University of California, San Francisco, National Institutes of Health, National Cancer Institute, Shanghai Public Health Clinical Center, Guangdong Medical College, Fudan University, ShenZhen People’s Hospital
Variable analysis
- Incubation time of BMDCs with beads
- Concentration of Alexa Fluor 647 (AF647)–conjugated OVA
- Uptake of AF647-OVA by splenic cDCs
- Uptake of beads by BMDCs
- Cell types used (splenic cDCs and BMDCs)
- Culture medium (RPMI 1640 complete medium)
- Temperature (37°C)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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