cd86 Search Results


cd86 antibody  (Miltenyi Biotec)
95
Miltenyi Biotec cd86 antibody
A) Workflow for assessing the presence of cellular proteins (CD63, ICAM1 and <t>CD86)</t> and virus-encoded therapeutic payloads (IL-12, CD40L and OVA peptide SIINFEKL presented on MHCI) on EVs secreted by mouse dendritic cells (DC2.4) and isolated by ultracentrifugation (UC) or size exclusion chromatography (SEC; EVs-rich fractions (EVs SEC, fractions 1-4) and soluble proteins-rich fractions (Prots SEC, fractions 5-10)) following 100nm filtration. Payloads were quantified by electrochemiluminescence immunoassay (B-G) on EVs isolated from non-infected cells (mock) or from cells infected with empty or armed MVA virus (encoding IL12, CD40L and OVA peptide SIINFEKL payloads). B) Quantification of CD63 on EVs. EVs mock Vs EVs empty virus p=0,0007; EVs mock Vs EVs armed virus UC p= 0,0001; EVs mock Vs EVs armed virus SEC p< 0,0001; EVs mock Vs Soluble proteins armed virus SEC p< 0,0001; One-way Anova; N=5 independent experiments. C) Quantification of ICAM1 on EVs. EVs mock Vs EVs empty virus: p=0,0276; EVs mock Vs EVs armed virus UC p= 0,0012; EVs mock Vs EVs armed virus SEC p= 0,0425; EVs mock Vs Soluble proteins armed virus SEC p= 0,83; One-way Anova; N=4 independent experiments. D) Quantification of CD86 on EVs. EVs mock Vs EVs empty virus: p=0,19; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs EVs armed virus SEC p= 0,73; EVs mock Vs Soluble proteins armed virus SEC p= 0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of CD40L on EVs. EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p<0,0001; EVs mock Vs EVs armed virus SEC p= 0,011; EVs mock Vs Soluble proteins armed virus SEC p>0,99; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,0125; One-way Anova; N=4 independent experiments. F) Quantification of MHCI bound OVA peptide (SIINFEKL). EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p=0,023; EVs mock Vs EVs armed virus SEC p= 0,0341; EVs mock Vs Soluble proteins armed virus SEC p>0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of IL-12. EVs mock Vs EVs empty virus p=0,045; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs Soluble proteins armed virus SEC p=0,0004; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,045; Kruskal-Wallis test. N=5 independent experiments. * p<0,05; ** p<0,01; *** p<0,005; **** p<0,001; ***** p<0,0001; ns: non-significant.
Cd86 Antibody, supplied by Miltenyi Biotec, 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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gene exp cd86 mm00444543 m1  (Thermo Fisher)
97
Thermo Fisher gene exp cd86 mm00444543 m1
Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and <t>CD86</t> mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.
Gene Exp Cd86 Mm00444543 M1, supplied by Thermo Fisher, 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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cd86  (Proteintech)
96
Proteintech cd86
Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and <t>CD86</t> mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.
Cd86, supplied by Proteintech, 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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cd86 cd206  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc cd86 cd206
Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and <t>CD86</t> mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.
Cd86 Cd206, supplied by Cell Signaling Technology Inc, 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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cd86 gl 1 pe  (Cytek Biosciences)
94
Cytek Biosciences cd86 gl 1 pe
Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and <t>CD86</t> mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.
Cd86 Gl 1 Pe, supplied by Cytek Biosciences, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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primary antibodies against cd86  (Proteintech)
95
Proteintech primary antibodies against cd86
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Primary Antibodies Against Cd86, supplied by Proteintech, 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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anti acly ptm 20018  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc anti acly ptm 20018
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Anti Acly Ptm 20018, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fitc anti mouse cd86  (Proteintech)
93
Proteintech fitc anti mouse cd86
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Fitc Anti Mouse Cd86, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti cd68  (Proteintech)
93
Proteintech anti cd68
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Anti Cd68, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti cd86 antibody  (Boster Bio)
99
Boster Bio anti cd86 antibody
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Anti Cd86 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti cd86  (Bio-Rad)
93
Bio-Rad anti cd86
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Anti Cd86, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rat anti cd86 po3  (Bio-Rad)
93
Bio-Rad rat anti cd86 po3
Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of <t>CD86</t> and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.
Rat Anti Cd86 Po3, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


A) Workflow for assessing the presence of cellular proteins (CD63, ICAM1 and CD86) and virus-encoded therapeutic payloads (IL-12, CD40L and OVA peptide SIINFEKL presented on MHCI) on EVs secreted by mouse dendritic cells (DC2.4) and isolated by ultracentrifugation (UC) or size exclusion chromatography (SEC; EVs-rich fractions (EVs SEC, fractions 1-4) and soluble proteins-rich fractions (Prots SEC, fractions 5-10)) following 100nm filtration. Payloads were quantified by electrochemiluminescence immunoassay (B-G) on EVs isolated from non-infected cells (mock) or from cells infected with empty or armed MVA virus (encoding IL12, CD40L and OVA peptide SIINFEKL payloads). B) Quantification of CD63 on EVs. EVs mock Vs EVs empty virus p=0,0007; EVs mock Vs EVs armed virus UC p= 0,0001; EVs mock Vs EVs armed virus SEC p< 0,0001; EVs mock Vs Soluble proteins armed virus SEC p< 0,0001; One-way Anova; N=5 independent experiments. C) Quantification of ICAM1 on EVs. EVs mock Vs EVs empty virus: p=0,0276; EVs mock Vs EVs armed virus UC p= 0,0012; EVs mock Vs EVs armed virus SEC p= 0,0425; EVs mock Vs Soluble proteins armed virus SEC p= 0,83; One-way Anova; N=4 independent experiments. D) Quantification of CD86 on EVs. EVs mock Vs EVs empty virus: p=0,19; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs EVs armed virus SEC p= 0,73; EVs mock Vs Soluble proteins armed virus SEC p= 0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of CD40L on EVs. EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p<0,0001; EVs mock Vs EVs armed virus SEC p= 0,011; EVs mock Vs Soluble proteins armed virus SEC p>0,99; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,0125; One-way Anova; N=4 independent experiments. F) Quantification of MHCI bound OVA peptide (SIINFEKL). EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p=0,023; EVs mock Vs EVs armed virus SEC p= 0,0341; EVs mock Vs Soluble proteins armed virus SEC p>0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of IL-12. EVs mock Vs EVs empty virus p=0,045; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs Soluble proteins armed virus SEC p=0,0004; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,045; Kruskal-Wallis test. N=5 independent experiments. * p<0,05; ** p<0,01; *** p<0,005; **** p<0,001; ***** p<0,0001; ns: non-significant.

Journal: bioRxiv

Article Title: Therapeutic poxviruses induce the secretion of immunostimulating and anti-tumoral extracellular vesicles

doi: 10.1101/2025.09.19.677320

Figure Lengend Snippet: A) Workflow for assessing the presence of cellular proteins (CD63, ICAM1 and CD86) and virus-encoded therapeutic payloads (IL-12, CD40L and OVA peptide SIINFEKL presented on MHCI) on EVs secreted by mouse dendritic cells (DC2.4) and isolated by ultracentrifugation (UC) or size exclusion chromatography (SEC; EVs-rich fractions (EVs SEC, fractions 1-4) and soluble proteins-rich fractions (Prots SEC, fractions 5-10)) following 100nm filtration. Payloads were quantified by electrochemiluminescence immunoassay (B-G) on EVs isolated from non-infected cells (mock) or from cells infected with empty or armed MVA virus (encoding IL12, CD40L and OVA peptide SIINFEKL payloads). B) Quantification of CD63 on EVs. EVs mock Vs EVs empty virus p=0,0007; EVs mock Vs EVs armed virus UC p= 0,0001; EVs mock Vs EVs armed virus SEC p< 0,0001; EVs mock Vs Soluble proteins armed virus SEC p< 0,0001; One-way Anova; N=5 independent experiments. C) Quantification of ICAM1 on EVs. EVs mock Vs EVs empty virus: p=0,0276; EVs mock Vs EVs armed virus UC p= 0,0012; EVs mock Vs EVs armed virus SEC p= 0,0425; EVs mock Vs Soluble proteins armed virus SEC p= 0,83; One-way Anova; N=4 independent experiments. D) Quantification of CD86 on EVs. EVs mock Vs EVs empty virus: p=0,19; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs EVs armed virus SEC p= 0,73; EVs mock Vs Soluble proteins armed virus SEC p= 0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of CD40L on EVs. EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p<0,0001; EVs mock Vs EVs armed virus SEC p= 0,011; EVs mock Vs Soluble proteins armed virus SEC p>0,99; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,0125; One-way Anova; N=4 independent experiments. F) Quantification of MHCI bound OVA peptide (SIINFEKL). EVs mock Vs EVs empty virus: p>0,99; EVs mock Vs EVs armed virus UC p=0,023; EVs mock Vs EVs armed virus SEC p= 0,0341; EVs mock Vs Soluble proteins armed virus SEC p>0,99; Kruskal-Wallis; N=4 independent experiments. E) Quantification of IL-12. EVs mock Vs EVs empty virus p=0,045; EVs mock Vs EVs armed virus UC p>0,99; EVs mock Vs Soluble proteins armed virus SEC p=0,0004; EVs armed virus SEC Vs Soluble proteins armed virus SEC p=0,045; Kruskal-Wallis test. N=5 independent experiments. * p<0,05; ** p<0,01; *** p<0,005; **** p<0,001; ***** p<0,0001; ns: non-significant.

Article Snippet: The following detection antibodies were used: CD63 Antibody, anti-mouse Biotin (clone REA563, Miltenyi Biotec); CD54 (ICAM-1) Antibody, anti-mouse, Biotin (clone YN1/1.7.4, #130-104-213, Miltenyi Biotec); CD86 Antibody, anti-mouse, Biotin (clone PO3.3, #130-101-944, Miltenyi Biotec); CD154 (CD40L) Antibody, anti-mouse, Biotin (clone MR1, #130-101-900, Miltenyi Biotec); H-2Kb/SIINFEKL Antibody, anti-mouse Biotin (clone 25-D1.16, Miltenyi Biotec).

Techniques: Virus, Isolation, Size-exclusion Chromatography, Filtration, Electrochemiluminescence, Infection

Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and CD86 mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.

Journal: Frontiers in Pharmacology

Article Title: The TLR4-Active Morphine Metabolite Morphine-3-Glucuronide Does Not Elicit Macrophage Classical Activation In Vitro

doi: 10.3389/fphar.2016.00441

Figure Lengend Snippet: Effect of M3G co-administered with IFN-γ on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were treated with 1 ng/ml IFN-γ alone or together with M3G (1, 5, 10, or 20 μM) for 12 h. Expression of iNOS, TNF-α, IL-6, and CD86 mRNA was determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Mean ± SEM is shown, n = 4 independent experiments. ∗ p < 0.05, M3G + IFN-γ vs. IFN-γ alone, one-way ANOVA analysis with Dunnett’s multiple comparisons. Results are shown as mean ± SEM, n = 4–7 independent experiments.

Article Snippet: Amplification and quantification of cDNA was assessed, using TaqMan TM Fast Universal PCR Master Mix (Life Technologies, VIC, Australia) with AmpliTaq Gold TM DNA Polymerase and TaqMan TM Gene Expression Assays including human primers : IL-6 (Hs00985639-m1), IL-12 (Hs01011518-m1), IL-23 (Hs00900828-g1), TNFα (Hs01113624-g1), CXCL10 (Hs01124252-g1), and CXCL11(Hs04187682-g1) or mouse primers: iNOS (Mm00440502-m1), IL-6 (Mm00446190-m1), TNFα (Mm00443258-m1), CD86 (Mm00444543-m1) in a StepOnePlus 7500 real time PCR system (Applied Biosystems, Carlsbad, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Control

Effect of M3G on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were exposed to 0.001 or 0.01 ng/ml LPS, or M3G at the indicated concentrations (1, 5, 10, and 20 μM). The mRNA levels of iNOS, CD86, IL-6, or TNF-α were determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Results are shown as mean ± SEM, n = 4–7 independent experiments. ∗∗ p < 0.01, LPS vs. control cells.

Journal: Frontiers in Pharmacology

Article Title: The TLR4-Active Morphine Metabolite Morphine-3-Glucuronide Does Not Elicit Macrophage Classical Activation In Vitro

doi: 10.3389/fphar.2016.00441

Figure Lengend Snippet: Effect of M3G on expression of M1 markers in RAW264.7 cells. RAW264.7 cells (2 × 10 5 cells/well) were exposed to 0.001 or 0.01 ng/ml LPS, or M3G at the indicated concentrations (1, 5, 10, and 20 μM). The mRNA levels of iNOS, CD86, IL-6, or TNF-α were determined by qRT-PCR. Results are shown relative to control (untreated) RAW264.7 cells. Results are shown as mean ± SEM, n = 4–7 independent experiments. ∗∗ p < 0.01, LPS vs. control cells.

Article Snippet: Amplification and quantification of cDNA was assessed, using TaqMan TM Fast Universal PCR Master Mix (Life Technologies, VIC, Australia) with AmpliTaq Gold TM DNA Polymerase and TaqMan TM Gene Expression Assays including human primers : IL-6 (Hs00985639-m1), IL-12 (Hs01011518-m1), IL-23 (Hs00900828-g1), TNFα (Hs01113624-g1), CXCL10 (Hs01124252-g1), and CXCL11(Hs04187682-g1) or mouse primers: iNOS (Mm00440502-m1), IL-6 (Mm00446190-m1), TNFα (Mm00443258-m1), CD86 (Mm00444543-m1) in a StepOnePlus 7500 real time PCR system (Applied Biosystems, Carlsbad, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Control

Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of CD86 and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.

Journal: Molecules (Basel, Switzerland)

Article Title: Inosine Prevents Colorectal Cancer Progression by Inducing M1 Phenotypic Polarization of Macrophages.

doi: 10.3390/molecules30010123

Figure Lengend Snippet: Figure 1. Inosine promoted macrophage polarization toward the M1 phenotype. (A) RAW264.7 cells were treated with inosine (0–40 mM) for 24 h; then, an MTT assay was performed to observe cell viability. RAW264.7 cells were administrated with inosine (1.25, 2.5, and 5 mM) in the absence or presence of LPS+IFN-γ or IL-4. (B,C) The proportions of CD86 and CD206-positive cells were determined by a flow cytometer. (D,E) The expression levels of CD86 and CD206 mRNA were detected by RT-qPCR. (F–H) Levels of CD86 and iNOS were measured by WB. Compared to the control group: * p < 0.05, ** p < 0.01, *** p < 0.001; Compared with the LPS+IFN-γ induced M1 group: # p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the IL-4 induced M2 group: ∆∆∆p < 0.001.

Article Snippet: Primary antibodies against CD86 (1:1000, 26903-1-AP, Proteintech, Wuhan, China), iNOS (1:300, 22226-1-AP, Proteintech, Wuhan, China), β-actin (1:2000, GB15003-100, Servicebio, Wuhan, China) and secondary antibody against HRP Goat Anti-Rabbit IgG (1:5000, AS014, ABclonal, Wuhan, China) were used, respectively.

Techniques: MTT Assay, Flow Cytometry, Expressing, Quantitative RT-PCR, Control

Figure 4. Effects of inosine on immune factors in the CT26 tumor microenvironment. (A) Effect of inosine on Ki-67 expression in tumor tissues (Scale: 100 µm; 400× and 200×). (B) Statistics of Ki-67 protein positive expression in tumor tissues (n = 3). (C) Fluorescence co-localization fluorogram of M1- type macrophage marker F4/80 + CD86 (scale: 100 µm; 200×). (D) F4/80 + CD86 expression statistics in tumor tissues. (E) M2 type macrophage marker F4/80 + CD206 fluorescence co-localization fluorogram (scale: 100 µm; 200×). (F) F4/80 + CD206 expression statistics in tumor tissues. Note: 5-Fu are 5-Fu (12 mg/kg) groups. IS-L and IS-H are inosine low and high-dose (5 mg/kg and 50 mg/kg) groups, respectively. Yellow arrows represent positive positions. Compared with the model group: ## p < 0.01; Compared with the 5-Fu group: △p < 0.05.

Journal: Molecules (Basel, Switzerland)

Article Title: Inosine Prevents Colorectal Cancer Progression by Inducing M1 Phenotypic Polarization of Macrophages.

doi: 10.3390/molecules30010123

Figure Lengend Snippet: Figure 4. Effects of inosine on immune factors in the CT26 tumor microenvironment. (A) Effect of inosine on Ki-67 expression in tumor tissues (Scale: 100 µm; 400× and 200×). (B) Statistics of Ki-67 protein positive expression in tumor tissues (n = 3). (C) Fluorescence co-localization fluorogram of M1- type macrophage marker F4/80 + CD86 (scale: 100 µm; 200×). (D) F4/80 + CD86 expression statistics in tumor tissues. (E) M2 type macrophage marker F4/80 + CD206 fluorescence co-localization fluorogram (scale: 100 µm; 200×). (F) F4/80 + CD206 expression statistics in tumor tissues. Note: 5-Fu are 5-Fu (12 mg/kg) groups. IS-L and IS-H are inosine low and high-dose (5 mg/kg and 50 mg/kg) groups, respectively. Yellow arrows represent positive positions. Compared with the model group: ## p < 0.01; Compared with the 5-Fu group: △p < 0.05.

Article Snippet: Primary antibodies against CD86 (1:1000, 26903-1-AP, Proteintech, Wuhan, China), iNOS (1:300, 22226-1-AP, Proteintech, Wuhan, China), β-actin (1:2000, GB15003-100, Servicebio, Wuhan, China) and secondary antibody against HRP Goat Anti-Rabbit IgG (1:5000, AS014, ABclonal, Wuhan, China) were used, respectively.

Techniques: Expressing, Fluorescence, Marker