All flow cytometry used appropriate isotype controls. Gating and compensation were aided by the performing fluorescence minus one controls. Cells isolated from biopsies or blood were stained with blue-fluorescent reactive dye (Life Technologies), CD19-BV785, CD27-APC (BD Biosciences) or PE or BV421, CD10-BV605 or APC, CD38-PerCp-ef710 (eBioscience), CD24-PE/Cy7 or BV605, IgD-APC/Cy7, IgM-V450 (BD Biosciences), IgG-PE/Cy7 and IgA-FITC (Miltenyi Biotec), in 2% FCS staining buffer with 1 mM EDTA for 15 min at 4 °C before analysis on the BD LSRFortessa (BD Biosciences). For high-throughput sequencing analysis, cells were stained with LIVE/DEAD Fixable Aqua (Life Technologies) or DAPI, CD19-BV785, CD27-FITC or APC, IgD-APC/Cy7, CD38-PerCp-ef710 (eBiosciences), CD10-BV605 in 2% FCS staining buffer with 1 mM EDTA for 15 min at 4 °C before sorted on the BD FACSAria (BD Biosciences). Four subsets from PBMC and three subsets from paired biopsy mononuclear cells from Donors 1 to 4. The numbers of cells used to generate sequences for each sample from Donors 1 to 4 are shown in Supplementary Fig. 14d.
Zhao Y., Uduman M., Siu J.H., Tull T.J., Sanderson J.D., Wu Y.C., Zhou J.Q., Petrov N., Ellis R., Todd K., Chavele K.M., Guesdon W., Vossenkamper A., Jassem W., D’Cruz D.P., Fear D.J., John S., Scheel-Toellner D., Hopkins C., Moreno E., Woodman N.L., Ciccarelli F., Heck S., Kleinstein S.H., Bemark M, & Spencer J. (2018). Spatiotemporal segregation of human marginal zone and memory B cell populations in lymphoid tissue. Nature Communications, 9, 3857.
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