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Cellular uptake and <t>tumor</t> <t>spheroid</t> penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and <t>3D</t> reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).
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Cellular uptake and <t>tumor</t> <t>spheroid</t> penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and <t>3D</t> reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).
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Cellular uptake and <t>tumor</t> <t>spheroid</t> penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and <t>3D</t> reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).
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Angio-Proteomie quick coating solution
Cellular uptake and <t>tumor</t> <t>spheroid</t> penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and <t>3D</t> reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).
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Angio-Proteomie matrigel coating solution
Cellular uptake and <t>tumor</t> <t>spheroid</t> penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and <t>3D</t> reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).
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Cellular uptake and tumor spheroid penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and 3D reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).

Journal: Bioactive Materials

Article Title: LRP-1/CD44-targeted regorafenib nano-delivery system leveraging anti-angiogenesis and synergistic cytotoxicity against peritoneal metastasis of colorectal cancer

doi: 10.1016/j.bioactmat.2025.12.015

Figure Lengend Snippet: Cellular uptake and tumor spheroid penetration. (A) Confocal microscopy images of CT26-luc cells after 2 h of incubation with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups pre-treated with free LF or HA. Scale bar: 50 μm. (B) Flow cytometry histograms of (i) CT26-luc cells, (ii) human umbilical vein endothelial cells (HUVECs), and (iii) M2-polarized bone marrow-derived macrophages (M2-BMDMs) treated with C6@LF NPs, C6@LFHA NPs, and competitive inhibition groups. (C) Lysosomal co-localization of C6@LFHA NPs in HUVECs after 8 h incubation. Quantitative analysis showed a Pearson's correlation coefficient of 0.34 ± 0.026. (D) Penetration of C6@LFHA NPs into CT26-luc tumor spheroids, as shown by Z-stack slices (Scale bar: 20 μm), maximum intensity projection, and 3D reconstruction (generated using ImageJ). Scale bar: 200 μm. All data are presented as mean ± SD (n = 3).

Article Snippet: For tumor spheroid penetration studies, CT26-luc multicellular tumor spheroids (MCTS) were prepared using a 3D cell culture coating kit (Beyotime).

Techniques: Confocal Microscopy, Incubation, Inhibition, Flow Cytometry, Derivative Assay, Generated