Journal: bioRxiv

Article Title: Senescent fibroblasts interfere with the tumor immune response in humanized models by inducing neutrophil extracellular traps

doi: 10.64898/2026.02.09.704891

Figure Lengend Snippet: A) Schematic illustration of the experimental workflow. NSG-SGM3 mice were injected intravenously (i.v) with 4x10 5 non-senescent or senescent HDF. The next day, 1x10 5 tumor cells (LEC-4T) were injected i.v.. Six days after tumor cell injection, mice were injected i.v with hCD45 + cells (5x10 6 PBMCs and 5x10 6 granulocytes each/mouse). Tumor growth was evaluated weekly until day 36, when mice were sacrificed and their lungs collected for analysis. B) Representative in vivo bioluminescence images of mice injected with luciferase-expressing LEC-4T and HDF stained with NIR790 dye. Also shown is a mouse without HDF. C) Graph showing the integrated density of HDF stained with NIR790 and tracked over time in LEC-4T tumors growing in NSG-SGM3 mice injected (dashed line) or not injected (solid line) with hCD45 + cells. Non-senescent (NS) HDF are shown in red, HDF exposed to IR in blue, and HDF expressing RAS in green. Data are presented as mean ± SEM from 8–9 tumors. D) Representative bioluminescence images of tumor-bearing mice with or without hCD45 + cells. Mice were injected with LEC-4T alone or with non-senescent or senescent HDF (IR or RAS). Images were captured on days 6, 13, and 36. E) Growth curves of LEC-4T tumors injected alone (black) or with non-senescent HDF (red), senescent HDF induced by irradiation (blue), or senescent HDF induced by RAS (green) in mice without hCD45 + cells. Each line represents the mean tumor growth ± SEM for 15 mice in each group over 36 days. Statistical analyses were performed using a mixed-effects model followed by Tukey’s multiple comparison test. **p < 0.01; ***P < 0.001. F) Tumor growth curves for individual mice injected with hCD45 + cells. LEC-4T tumor cells injected alone are shown in black (n = 6), with non-senescent HDF in red (n = 7), with senescent HDF induced by irradiation in blue (n = 8), or with senescent HDF induced by RAS in green (n = 7). Also shown is a pie chart illustrating the mean percentage of mice that achieved complete tumor elimination (CE) after immune reconstitution.

Article Snippet: Non-senescent or senescent HDF cells were stained with the fluorescent dye CellBrite NIR790 Cytoplasmic Membrane Dye (catalogue 30079; Biotium) following the manufacturer’s instructions before being injected into mice.

Techniques: Injection, In Vivo, Luciferase, Expressing, Staining, Irradiation, Comparison