Journal: bioRxiv
Article Title: Phenotypic dynamics and behaviors of neutrophils during barrier challenge
doi: 10.64898/2026.01.06.697946
Figure Lengend Snippet: A) Histogram representations of CD101 expression in neutrophils from 4h, 12h, 24h, and 48h infections. B) Histogram representations of CD101 expression in neutrophils from i.v.+ versus i.v.- compartments of h, 12h, 24h, and 48h infections. C) Schematic representation of EdU experimental design. D) Quantification of flow cytometry data depicting EdU positivity as a proportion of various neutrophil subsets. From left to right, CD101-lo bone marrow neutrophils (blue), CD101-hi bone marrow neutrophils (pink), blood neutrophils (purple), and CD101-hi (orange) or CD101-lo (green) skin infection-associated neutrophils. N = 4-5 mice per data point. E) Schematic depiction of adoptive transfer scheme, wherein blood neutrophils of CD45.1 congenic mice were isolated and transferred into infection sites of CD45.2 recipient mice, and analyzed 4h later for phenotypic traits. I) Histogram representations of flow cytometry data depicting CD101 expression in CD45.1 donor neutrophils (top, teal) and CD45.1 neutrophils recovered 4h after adoptive transfer (lower, red), each red histogram representing cells from an individual recipient mouse. J) Quantification of the proportion of endogenous CD45.2 (gray) and donor-derived CD45.1 (red) neutrophils falling into the CD101-lo gate. K) Schematic representation of analysis to compare CD101-hi to CD101-lo i.v.- neutrophils from either bone marrow or skin infections. L-M) Volcano plot representation of phenotypic markers and how they differ between CD101-lo and CD101-hi populations in bone marrow (L) and skin infections (M). Delta gMFI values represent averages over N = 5 mice. N) Venn-diagram representing markers that are decreased (blue) or increased (red) in expression within the CD101-lo gate, compared to the CD101-hi gate, from each tissue. O-P) Pair-wise comparisons of PD-L1 (O) or ICAM-1 (P) expression in CD101-hi versus CD101-lo subpopulations of neutrophils from bone marrow (left) and skin infections (middle), and representative flow cytometry data depicting PD-L1 expression in infection associated neutrophils (right). Error bars = SEM, statistics = t-test for P values shown in volcano plots.
Article Snippet: Spectral flow cytometry data was collected on a Cytek Aurora (5-laser) nad spillover/compensation was adjusted in FlowJo before export to omiq.ai for further analysis.
Techniques: Expressing, Flow Cytometry, Infection, Adoptive Transfer Assay, Isolation, Derivative Assay