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. 2020 Aug 25;11:4249. doi: 10.1038/s41467-020-17996-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Efficacy study in subcutaneous PANC-1 xenograft model. Tumor bearing mice (n = 4) received 5 i.v. injections of co-delivery NP (210 mg particles/kg/injection), as well as single drug-laden nanoparticle or free drugs at an equivalent PAL dose of 10 mg/kg/injection and an HCQ dose of 33 mg/kg/injection. Mice treated with saline were also studied as control. The tumor size was monitored for 21 days from the first injection and plotted versus time. Co-delivery NP led to the most potent tumor inhibition compared to all the controls, e.g., co-delivery NP versus Free PAL + HCQ (p = 7 × 10⁻⁵). Representative images of tumor harvested on day 21 further demonstrated the significantly enhanced antitumor efficacy by co-delivery NP. b Confirmative biodistribution study in orthotopic PANC-1 xenograft model. Mice (n = 3) received single i.v. injection of DyLight680-labeled co-delivery NP (210 mg particles/kg) and sacrificed 48 h post injection. The orthotopic tumor site was localized by bioluminescence-based imaging. Ex vivo IVIS imaging were performed to determine the NIR signals from major organs and tumor to identify the distribution of nanoparticles. c Confocal microscopy confirmed the abundance of DyLight680-labeled co-delivery NP (red) in the section of orthotopic tumor stained with anti-CD31 antibody (green) and DAPI (blue). Scale bar: 100 μm. d Efficacy study in orthotopic PANC-1 model. Mice (n = 4) received the same dosage regimen as in a. Live-animal IVIS imaging was performed to monitor the orthotopic tumor growth for 30 days from the first injection and the representative bioluminescence is shown. The bioluminescence intensity was quantified at the ROI by the IVIS Living Image software v4.5 and plotted versus time. Co-delivery NP outperformed all the control groups, e.g., co-delivery NP versus Free PAL + HCQ (p = 0.025). e Immunohistochemical staining of tumor sections harvested on day 30 revealed significant inhibition of pRb and Ki67. Scale bar: 100 μm. Data were obtained from one experiment without repetition. Data represent mean ± SD; statistical difference was evaluated by one-way ANOVA followed by Tukey’s post hoc test (*p < 0.05). Source data are provided as a Source data file.