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. Author manuscript; available in PMC: 2024 Jun 9.
Published in final edited form as: Nat Biomed Eng. 2024 Apr 19;8(5):593–610. doi: 10.1038/s41551-024-01194-7

Fig. 1. Physiochemical characterization and antitumor evaluation of polypeptides in vivo.

Fig. 1.

(a) Schematic illustration and chemical structure of the P1, P2, and P3 polypeptides. (b) Representative fluorescence images of EO771 tumor-bearing mice taken at predetermined times after intravenous injection of IR800CW-tagged P1, P2, or P3 (10 mg/kg), and ex vivo fluorescence images of tumor and major organs harvested at 24 h after administration. (c) Fluorescence images of EO771 tumor tissues excised at 24 h after treatment with IR800CW-tagged P1, P2 or P3. (d) Quantification of total radiation efficiency (near infrared fluorescence signal) of tumor and major organs at 24 h after treatment (n=3, mean±standard deviation, SD); unpaired Student’s t test in comparison with P1. (e) Time course of polypeptide concentrations in blood plasma after intravenous administration of IR800CW-tagged P1, P2, or P3 (10 mg/kg, n=3, mean±SD). Half-life and area under the curve (AUC) values were calculated by GraphPrism. (f, g) Measurement of IR800CW+ fluorescence signals in (f) CD45+ cells (leukocytes), CD45− EpCAM+ (EO771 tumor cells), and (g) immune cell subtypes (macrophages: CD11b+CD11c−F4/80+, dendritic cells [DCs]: CD11c+MHC-II+F4/80−, neutrophils: CD11b+CD11c−MHC-II−Gr1+, T cells: CD3+, NK cells: CD3−NKp46+NK1.1+) by flow cytometry of tumors 24 after treatment with IR800CW-tagged P1, P2, or P3 (10 mg/kg). Unpaired two-tailed Student’s t test in comparison with CD45+ for (f) or to P1 for (g). (h) Immunofluorescence images show that IR800CW-tagged P1 accumulates in macrophages and DCs within tumor microenvironments to a greater extent than IR800CW-tagged P2 or P3; scale bar, 15 μm (i) Timeline of treatment with P1, P2, and P3 in EO771 tumor-bearing mice. (j) P1 suppressed tumor growth and extended survival of tumor-bearing mice compared with P2 or P3 (n=5, mean±SD), unpaired Student’s t test in comparison with P1 at day 28 after tumor inoculation for tumor growth curves; log-rank (Mantel-Cox) test for Kaplan-Meier survival curves.