Journal: bioRxiv
Article Title: Cancer-driven neutrophil priming couples systemic epithelial regenerative programs with pre-metastatic niche formation
doi: 10.1101/2025.10.20.683483
Figure Lengend Snippet: a) Pathways enriched in mature bone marrow (BM) neutrophils from tumour bearing (TB) versus Naïve mice. Inflammatory and metabolic pathways are coloured in red and blue, respectively; enrichment P values are indicated. b) LC–MS–based metabolic profiling of control or neutrophil-conditioned media. Neutrophils were isolated from lung or spleen of naïve or TB (4T1 Balb/c) mice (n=4 per group) and incubated in Advanced DMEM/F12 medium. Data are shown as mean z-score per metabolite. c-d) Abundance of uridine (c) and uracil (d) in neutrophil-conditioned or control media generated from lung or splenic neutrophils isolated from naïve or TB (4T1 Balb/c) mice, incubated in uridine-containing Plasmax medium (n=4 mice per group), ordinary one-way ANOVA. e) Schematic representation of Uridine phosphorylase 1 (Upp1) enzymatic activity. f) Upp1 mRNA expression in lung and splenic neutrophils from naïve or TB (PyMT-FVB) mice lung (n=5 per group, spleen n=4 per group), two-tailed students t-test. g) (Left) representative image and (Right) quantification of UPP1 immunofluorescence intensity in lung neutrophils (LuNeu) isolated from naïve or tumour bearing (PyMT-FVB) mice. Data are represented a per cell mean intensity (n=4). Welch’s t-test. h) Plasma Uracil abundance from Naïve (Balb/c) IgG, TB (4T1) IgG or TB (4T1) αLy6G mice measured by LC-MS (Naïve IgG and TB IgG n=10, TB αLy6G n=5), ordinary one-way ANOVA. i) Upp1 expression in circulating neutrophils from healthy volunteers (HV) and patients with breast cancer (P). Data are normalized counts from bulk RNA-seq of purified neutrophils (n=14 per group), two-tailed Wilcoxon test. Hormone-receptor status of patients is indicated. j) Plasma uracil concentrations in matched samples from HV and P with breast cancer (n=17 per group), quantified by LC–MS, two-tailed Wilcoxon test.
Article Snippet: Primary antibody Upp1 (Proteintech 14186-1-AP, 1:200) in IF Wash Buffer 1% BSA was incubated overnight at 4 °C prior to washing with PBS.
Techniques: Liquid Chromatography with Mass Spectroscopy, Control, Isolation, Incubation, Generated, Activity Assay, Expressing, Two Tailed Test, Immunofluorescence, Clinical Proteomics, RNA Sequencing, Purification