rs504393  (Tocris)

Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Immune regulation and endogenous bone regeneration mechanism investigation. A) Network diagram showing the number of interactions between six subclusters. B) KEGG enrichment analysis of the upregulated DEGs in DIBS group compared to the HA group. C) Circular visualization of related pathway–gene enrichment analysis. D) Heatmap of key gene regulation in specific pathways. E) qRT-PCR validation for key gene expression in specific pathways. F) The interaction networks showing the correlation of representative immunomodulatory genes (CCL2, CCL20, Sfrp1, and Stat3, etc.) with angiogenesis/osteogenesis and macrophage regulation gene sets. G) Flow cytometry analysis and quantification of CCR2 F4/80 macrophage in peripheral blood. H) Immunofluorescence staining analysis of macrophage polarization inside scaffolds (one week after intramuscular implantation). I) Macrophage proliferation assay in a CCR2-dependent manner. J and K) Macrophage polarization assay in a CCR2-dependent manner. Data are represented as means ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Gene Expression, Flow Cytometry, Immunofluorescence, Staining, Proliferation Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration

doi: 10.1016/j.bioactmat.2026.04.002

Figure Lengend Snippet: Revascularization and osteogenesis are reinforced by M2 macrophage activation via the CCL2/CCR2 pathway. A and B) HUVECs and BMSCs proliferation assay under M2 macrophage activation. Created with BioRender.com . C) Migration assay and quantification of HUVECs. D) Tube formation assay and quantification of HUVECs. E and F) Early and later osteogenic differentiation of BMSC influenced by macrophage-induced microenvironment. Data are represented as means ± SD, ∗ p < 0.05 (vs Control), ∗∗ p < 0.01 (vs Control), ∗∗∗ p < 0.001 (vs Control), ∗∗∗∗ p < 0.0001 (vs Control); $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.

Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).

Techniques: Activation Assay, Proliferation Assay, Migration, Tube Formation Assay, Control

Journal: Advanced Science

Article Title: CCN1 Enhances Tumor Immunosuppression through Collagen‐Mediated Chemokine Secretion in Pancreatic Cancer

doi: 10.1002/advs.202500589

Figure Lengend Snippet: Collagens modulate chemokine expression in PDAC cells under IFNγ stimulation. A) Tumor growth of sgCtrl and sgCcn1 KPC cells inoculated subcutaneously in C57B/L6 mice. Tumor‐bearing mice were treated with CCR2 inhibitor RS504393 and CXCR2 inhibitor SB225002. n = 6 mice per group. B) Representative images of tumors (left) and quantification of tumor weight (right) in mice inoculated with sgCtrl or sgCcn1 KPC cells. Tumor‐bearing mice were treated with the CCR2 inhibitor RS504393 and the CXCR2 inhibitor SB225002. C) Flow cytometric analysis of MDSC proportions in subcutaneous sgCtrl and sgCcn1 KPC tumors. D) mRNA levels of collagens, including Col4a1, Cola4a2, Col5a1, Col6a1, Col6a2, Col6a3, and Col8a1, in KPC cells after their total knockdown. E) mRNA levels of chemokines, including Ccl2, Ccl20, Cxcl1, Cxcl3, Cxcl5, Csf1, Csf2, and Csf 3 , in KPC cells after their total knockdown. F–H) mRNA levels of chemokines, including Ccl2, Ccl7, Ccl20, Cxcl1, Cxcl3, Cxcl5, Csf1, Csf2, and Csf3, in KPC cells after Col5a1 knockdown (F), Col6a3 knockdown (G), and Col8a1 knockdown (H). I) mRNA levels of collagens, including Col4a1, Cola4a2, Col5a1, Col6a1, Col6a2, Col6a3, and Col8a1, in sgCtrl KPC cells after treatment with 100 ng mL −1 IFNγ. J) mRNA levels of Col5a1, Col6a3, and Col8a1 measured in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ. K–M) mRNA levels of chemokines including the Ccls (K), Cxcls (L), and Csfs (M), in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ. N) mRNA level of Il6 in siCol5a1, siCol6a3, and siCol8a1 KPC cells after treatment with 100 ng mL −1 IFNγ.

Article Snippet: CCR2 inhibitor RS504393 (S9738, Selleck), CXCR2 inhibitor SB225002 (S7651, Selleck), and gemcitabine (S1714, Selleck) were purchased from Selleckchem.

Techniques: Expressing, Knockdown

Journal: Advanced Science

Article Title: CCN1 Enhances Tumor Immunosuppression through Collagen‐Mediated Chemokine Secretion in Pancreatic Cancer

doi: 10.1002/advs.202500589

Figure Lengend Snippet: Ccn1 regulates the TME in PDAC. A) Schematic representation of macrophage isolation from mouse bone marrow. B) Macrophage invasion evaluated by the chemotactic effect of sgCtrl and sgCcn1 KPC cells in a Transwell invasion assay. C) Macrophage polarization assessed by flow cytometry in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. D) Macrophage polarization assessed by flow cytometry in macrophages cocultured with sgCtrl and sgCcn1 KPC cells after treatment with CCR2 inhibitor RS504393 and CXCR2 inhibitor SB225002. E) Survival of sgCtrl and sgCcn1 KPC cells cocultured with macrophages, assessed by CCK‐8 assay. F) Cell death of sgCtrl and sgCcn1 KPC cells cocultured with macrophages, assessed by flow cytometry. G) Protein expression levels of TNFα in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. H) mRNA expression levels of TNFα in macrophages cocultured with sgCtrl and sgCcn1 KPC cells. I) Protein expression level of c‐Caspase 3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 TNFα. J) Protein expression level of c‐Caspase 3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 TNFα, mouse CCN1 protein, and cilengitide (αVβ3 and αVβ5 integrin inhibitor). K) Nucleus localization of CCN1 in KPC cells with CCN1 (red) and DAPI (blue) fluorescence. L) Protein expression level of p‐STAT3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ. M) mRNA levels of Ccl7, Cxcl3, and Csf3 in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ. N) mRNA level of Il6 measured in sgCtrl and sgCcn1 KPC cells after treatment with 100 ng mL −1 INFγ.

Article Snippet: CCR2 inhibitor RS504393 (S9738, Selleck), CXCR2 inhibitor SB225002 (S7651, Selleck), and gemcitabine (S1714, Selleck) were purchased from Selleckchem.

Techniques: Isolation, Transwell Invasion Assay, Flow Cytometry, CCK-8 Assay, Expressing, Fluorescence