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Beyotime proliferation
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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Beyotime edu assay kit
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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Beyotime ki67 cell proliferation detection kit
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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Beijing Solarbio Science cck 8 reagent
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
Cck 8 Reagent, supplied by Beijing Solarbio Science, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Beijing Solarbio Science cck 8 assay kit
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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Beyotime beyoclick edu cell proliferation assay kit
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
Beyoclick Edu Cell Proliferation Assay Kit, supplied by Beyotime, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Beyotime edu incorporation assay
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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MedChemExpress edu detection kit
IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage <t>proliferation</t> evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).
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IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage proliferation evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage proliferation evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).

Article Snippet: Cell proliferation was observed by laser confocal microscope at 24 h, 48 h, 72 h. Briefly, the proliferation of cells was assessed by BeyoClick EdU-488 Cell Proliferation Kit (Beyotime) according to the manufacturer’s instructions.

Techniques: Migration, Control, Expressing, Analysis, EdU Assay, Fluorescence, Staining, Transwell Migration Assay, Transwell Assay