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Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) <t>Enzyme-linked</t> <t>immunosorbent</t> <t>assay</t> <t>(ELISA)</t> quantification <t>of</t> <t>TGF-β1</t> and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).
Mouse Tgf β1 Elisa Kit, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) <t>Enzyme-linked</t> <t>immunosorbent</t> <t>assay</t> <t>(ELISA)</t> quantification <t>of</t> <t>TGF-β1</t> and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 <t>and</t> <t>TGF-β</t> in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.
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Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

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: Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: In Vitro, Flow Cytometry, Cell Tracking Assay, Labeling, Co-culture Assay, Control, Enzyme-linked Immunosorbent Assay

Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

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: Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: Quantitative RT-PCR, Analysis, Marker, Gene Expression, Derivative Assay, Isolation, RNA Extraction, Sequencing, Functional Assay, Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

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:

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: Cell-Signaling, Recombinant, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Software

Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 and TGF-β in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.

Journal: Frontiers in Immunology

Article Title: Shared immune perturbations in hepatocellular carcinoma-depression comorbidity and synergistic immunotherapeutic effects of fluoxetine combined with anti-PD-1

doi: 10.3389/fimmu.2026.1909503

Figure Lengend Snippet: Immune cell infiltration landscape and correlation between hub genes and M2 macrophage-mediated immunosuppression in HCC-depression comorbidity. (A) Box plots of immune cell infiltration proportions comparing normal vs tumor, control vs depression cohorts. (B) Representative flow cytometry dot plots for peripheral CD206 + M2 macrophages from each treatment group. (C) Correlation heatmap of GLCE/RBCK1/BTN3A3 expression with multiple immune cell subsets. (D) ELISA quantification of immunosuppressive cytokines IL-10 and TGF-β in mouse serum compared with the model group, * p < 0.05 or ** p < 0.01. n=3.

Article Snippet: Commercial mouse IL-10 (Cat. No. E-EL-M0046) and TGF-β (Cat. No. E-EL-M0051) ELISA kits were purchased from Elabscience (Wuhan, China).

Techniques: Immune Cell, Control, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay