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. 2023 Feb 11;14:771. doi: 10.1038/s41467-023-36321-6

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© The Author(s) 2023

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. 1 — a mRNA expression profiling by qRT-PCR in bone marrow-derived macrophages (BMDMs) after 1 µM CDNP-R848 or CDNP vehicle treatment Expression of Nos2, Il-6, tnfα, Il-1ß, lcn2 and Il-12 was assessed. (a–c: n = 3 BMDM cultures per group from three mice). b Protein expression profiling by flow cytometry in BMDMs after CDNP-R848 or CDNP vehicle treatment. Quantification of CD80, CD64 and CD38, CD71, CD172, and MerTK. Results are normalized to non-treated cells (NT) BMDMs. c Quantification of ROS production in BMDMs 24 h after CDNP-R848 (1 µM) or CDNP vehicle treatment as measured by flow cytometry after incubation with the probe CellROX Green. Non-treated (NT) cells served as negative control. d Gl261 glioma cells were co-cultured with BMDMs upon CDNP-R848, CDNP vehicle or LPS treatment. Cytotoxicity was quantified by LDH release. Values are corrected for spontaneous effector and target cell LDH release. n = 3 to 4 BMDM cultures per group from three to four mice per time point. e Representative histological image from three biological replicates (n = 3 mice) shows NP accumulation in the TME of Gl261 glioma but not in the adjacent healthy brain. Dashed line indicates glioma border. Micrograph of whole-mounted spleen shows NP uptake in splenic phagocytes. Scale bar is 100 µm. f, g Biodistribution analysis of CDNP-R848-VT680 within myeloid cells 24 h after a single, intravenous dose as assessed by flow cytometry of monocyte-derived myeloid cells (CD45⁺ CD11b^high, Ly6c⁺), tissue-resident myeloid cells (CD45⁺ CD11b^high, F4/80⁺), neutrophil like myeloid cells (CD45⁺ CD11b^high, CD11c^high), NK cells (NK1.1⁺), T-cells (CD3⁺) and microglia (CD45⁺ CD11b^intermediate) in the tumor-bearing hemisphere, contralateral hemisphere and blood. n = 5 CDNP-R848-VT680 mice and 4 PBS-treated animals. Data are representative of two independent experiments and presented as mean ± SEM. Statistical significance was determined by two-way ANOVA with Sidak’s test. n.a.: not applicable. NT: non-treated. FC: fold change.