Protocol detail

Profiling Listeria-Associated Immune Cells

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Antibodies specific for CD16/CD32 (93), CD45 (30-F11), CD11c (N418), CD11b (M1/70), Ly6G (1A8-Ly6g), B220 (RA3-6B2), MHC-II (M5/114.15.2), CD3 (145-2C11), F4/80 (BM8), CD19 (eBio1D3) from eBioscience; Ly6C (HK1.4) and CD103 (2E7) from BioLegend were used. Data were acquired using an iCyt Synergy and analyzed with FlowJo (Tree Star). The percentage of Listeria-associated (GFP⁺) cells in each population was determined by using cells from mice infected with L. monocytogenes SD2001 as a negative gating control in each experiment as described previously (18 (link),19 (link)).

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Jones G.S., Smith V.C, & D’Orazio S.E. (2017). Listeria monocytogenes replicate in bone marrow-derived CD11c+ cells, but not in dendritic cells isolated from the murine gastrointestinal tract. Journal of immunology (Baltimore, Md. : 1950), 199(11), 3789-3797.

Publication 2017

CD11c (N418), Ly6C (HK1.4) CD103 (2E7) iCyt Synergy

Antibodies Cd103 Cd11b Cells Listeria Mice Tree

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Variable analysis

independent variables

Listeria-associated (GFP⁺) cells

dependent variables

Percentage of Listeria-associated (GFP⁺) cells in each population

control variables

Cells from mice infected with L. monocytogenes SD2001 used as negative gating control in each experiment

controls

Positive control: Cells from mice infected with L. monocytogenes SD2001
Negative control: Cells from mice infected with L. monocytogenes SD2001 used as negative gating control

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