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We combine color-coding and sorting on a given attribute to reinforce the perception of grouped leaves of similar values.
On specified days of co-culture, cells were recovered from the culture dishes for analysis and sorting on a flow cytometer.
Flow cytometric analysis was performed on a FACSCalibur or LSRFortessa and sorting on a FACSAria I (both Becton Dickinson, BD, Franklin Lakes, NJ) and data were analyzed with FlowJo Tree Starr, Inc., Ashland, OR).
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Cells were analyzed and sorted on a FACS ARIA II (BD Biosciences).
After transfection, cells were stained for MHC I surface expression levels using PE-conjugated anti-MHC I mAb (W6/32, AbD Serotec) and sorted on a FACSAriaII (BD Biosciences).
For cell sorting by flow cytometry, samples were analysed and sorted on a BD FACSVantage SE (BD Biosciences).
EPLMs were stained and sorted on a FACSAria flow cytometer (Becton Dickinson) as previously described (Balciunaite et al., 2005).
Cells were analyzed on a three laser LSRII flow cytometer using DiVa software and sorted on a three laser FACSAria (BD Biosciences, Mountain View, CA).
Lymph node cells were stained with DAPI and antibodies to CD4 and CD8 and sorted on a MoFlo cell sorter (Beckman Coulter).
Lineage-depleted (Lin−) FL cells were stained with c-Kit-PE and Sca1-FITC (eBioscience) and sorted on a FACS DiVa (BD Biosciences) and analyzed with Cell Quest software (BD Biosciences).
For in vitro experiments, conventional (CD4+CD25−6.5+) or regulatory (CD4+CD25+6.5+) T cells specific for HA were stained with anti-CD4, anti-CD25 and 6.5 antibodies and sorted on a Facs Aria (BD Biosciences).
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