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Figure Lengend Snippet: Characterization of mouse tumor model using bioluminescence imaging, flow cytometry and histology. (A) Representative BLI images show an orthotopic 4T1 tumor in mice and its progression overtime. After surgical resection of the primary tumor 14 days after inoculation, metastatic outgrowth and recurrence is observed in the thoracic and abdominal regions. Quantification of the BLI signal from (B) the primary tumor and (C) thoracic region is shown, respectively (n=5). The 4T1 cancer cells (GFP reporter) and CD45+ leukocytes were quantified using flow cytometry in the (D) primary tumor, (E) liver and (F) lungs. (G) Histological evaluation depicts the progression of metastatic disease in the liver and lungs (20x magnification; scale bar = 100 μm; blue: nuclear stain; green:4T1 cancer cells). Mean ± SEM are plotted with statistics by one-/two-way ANOVA with a post hoc Tukey or Sidak’s test. **P<0.01; ***P<0.001; ****P<0.0001.

Article Snippet: The 4T1 model was established in BALB/c mice (Jackson Laboratories).

Techniques: Imaging, Flow Cytometry, Staining

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Figure Lengend Snippet: Quantification of nanoparticle uptake by APCs across different TMEs. (A) Timelines show animal modeling, nanoparticle administration and flow cytometry analysis. (B) Flow cytometry analysis of nanoparticle uptake by 4T1 cancer cells and CD45 + leukocytes in primary tumor. Flow cytometry analysis of nanoparticle uptake by CD11c + dendritic cells and F4/80 + macrophages in (C) primary tumor, (D) lungs at a stage of early metastasis, (E) liver at a stage of early metastasis, (F) lungs at a stage of late metastasis, and (G) liver at a stage of late metastasis (n=5 mice per group). Box and whisker plots (5–95 percentile) with statistics by one-/two-way ANOVA with a post hoc Tukey or Sidak’s test. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.

Article Snippet: The 4T1 model was established in BALB/c mice (Jackson Laboratories).

Techniques: Flow Cytometry, Whisker Assay

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Figure Lengend Snippet: Response of good responders to a tumor rechallenge. The treatment scheme of animals bearing 4T1 tumors included surgical resection of the primary tumor, anti-PD1 and untargeted immunostimulatory NP before and after surgery, indicated as mPEG-NP(x2), or only before surgery indicated as mPEG-NP(x1). The good responders from the two groups were rechallenged 42 days after the initial inoculation with 1×10 5 4T1 cells on their right flank. (A) Caliper measurement of the flank tumor size. (B) Kaplan-Meier survival analysis. N=4 mice per condition. Box and whisker plots (5–95 percentile) both with statistics by one-/two-way ANOVA with a post hoc Tukey or Sidak’s test. **P<0.01.

Article Snippet: The 4T1 model was established in BALB/c mice (Jackson Laboratories).

Techniques: Whisker Assay