Multiparametric Flow Cytometry Analysis

Cell suspensions from mouse liver, BM (one femur and tibia), spleen, thymus, mesenteric lymph nodes, and peritoneal cavity were processed as previously described (47 (link)), and data were acquired on a FACSCanto10c (BD) and analyzed with FlowJo Software (Tree Star).
The following anti-mouse antibodies were used: B220 (RA3-6B2, BioLegend), BAFFR (7H22-E16,BD), BP-1 (6C3, BioLegend), CD3e (145-2C11, eBioscience), CD4 (GK1.5, eBioscience), CD5 (53-7.3, BioLegend), CD8 (53-6.7, BioLegend), CD9 (MZ3, BioLegend), CD11b (M1/70, eBioscience), CD19 (6D5, 1D3, BioLegend), CD21/CD35 (7E9, BioLegend), CD23 (B3B4, BioLegend), CD24 (M1/69, BioLegend), CD25 (PC61, Biolgend), CD43 (S7, BioLegend), CD44 (IM7, BioLegend), CD45.1 (A20, BioLegend), CD45.2 (104, BioLegend), CD69 (H1.2F3, BioLegend), CD80 (16-10A1, BioLegend), CD86 (GL1, BioLegend), CD93 (AA4.1, BioLegend), DAPI (BIOTIUM), Gr-1 (RB6-8C5, eBioscience), IgDa (AMS-9.1, BioLegend), IgD (11-26c, BioLegend), IgMa (DS-1, MA-69, BioLegend), IgM (Il/41, RMM-1, BioLegend), TCRγδ (GL3, BioLegend), Live dead dye (Zombie Aqua Dye, BioLegend), Live dead dye (Zombie NIR, BioLegend), pERK (4B11B69, BD), pPLCγ2 (K86-1161, BD), Lin28b (AP1485C, ABGENT), and anti-rabbit IgG (BioLegend).

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Xu X., Deobagkar-Lele M., Bull K.R., Crockford T.L., Mead A.J., Cribbs A.P., Sims D., Anzilotti C, & Cornall R.J. (2020). An ontogenetic switch drives the positive and negative selection of B cells. Proceedings of the National Academy of Sciences of the United States of America, 117(7), 3718-3727.

Publication 2020

Anti antibodies Anti igg Cd11b Cd25 Cd35 Cd43 Cd44 Dapi Femur Igda Liver cell Lymph nodes Mesenteric Mouse Peritoneal cavity Rabbit Spleen Tcrγδ Thymus Tibia Tree

Corresponding Organization : MRC Human Immunology Unit

Other organizations : MRC Weatherall Institute of Molecular Medicine

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Variable analysis

independent variables

Cell suspensions from mouse liver, BM (one femur and tibia), spleen, thymus, mesenteric lymph nodes, and peritoneal cavity

dependent variables

Data acquired on a FACSCanto10c (BD) and analyzed with FlowJo Software (Tree Star)

control variables

Cells were processed as previously described (47 (link))

controls

No positive or negative controls were explicitly mentioned in the input.

Annotations

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