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mouse monoclonal antibody against human cd163  (Bio-Rad)


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    Bio-Rad mouse monoclonal antibody against human cd163
    Upadacitinib promotes monocyte-derived macrophages with an anti-inflammatory gene expression and functional profile A Schematic representation of the experiments. Monocytes were exposed to 10–100 nM Upadacitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Right, immunoblot analysis of pSTAT5, STAT5, pERK, ERK and pp38 by monocytes treated for 1 h to 100 nM Upadacitinib (Upa) and exposed to GM-CSF for the indicated time points. B Number of annotated genes whose expression is regulated in GM-MØ after 7d of Upadacitinib treatment (adj p < 0.05). C Volcano plot of RNAseq results showing the 100 nM Upadacitinib-induced gene expression changes in GM-MØ. D PCA analysis of GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ.Three independent donors are identified as I, II and III. E GSEA on the ranked comparison of the GM-MØ versus 10Upa-GM-MØ and GM-MØ versus 100Upa-GM-MØ transcriptomes, using the genes significantly modulated by GM-CSF (GM-MØ-specific markers) and M-CSF (M-MØ-specific markers) as data set. Normalized Enrichment Score (NES) and False Discovery Rate (FDRq) are indicated. F Relative expression of the indicated genes as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . G Production of activin A, IL-10 and LGMN by GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 8 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test; F = 92.63 for Activin A, F = 17.85 for IL-10, F = 33.39 for LGMN). H Immunoblot analysis of <t>CD163</t> and FOLR2 (down) by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and monocytes differentiated with M-CSF (M-MØ). In panels A-G, vinculin or GAPDH protein levels were determined as protein loading controls and a representative experiment of two independent donors is shown. I Phagocytic activity in GM-MØ, 100Upa-GM-MØ and M-MØ . Mean ± SEM of 5 independent donors are shown (* p < 0.05, one-way ANOVA with Tukey’s post hoc test, F = 13.74). J Production of TNFα, IL-6 and IL-10 by GM-MØ and 100Upa-GM-MØ challenged with LPS for 24 h, as determined by ELISA. Mean ± SEM of 7–8 independent donors are shown (* p < 0.05, ** p < 0.01, paired t-test)
    Mouse Monoclonal Antibody Against Human Cd163, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+monoclonal+antibody+against+human+cd163/Mouse+anti+Human+CD163/pmc10963568-76-31-39
    Average 94 stars, based on 237 article reviews
    mouse monoclonal antibody against human cd163 - by Bioz Stars, 2026-09
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    1) Product Images from "Macrophage re-programming by JAK inhibitors relies on MAFB"

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB

    Journal: Cellular and Molecular Life Sciences

    doi: 10.1007/s00018-024-05196-1

    Upadacitinib promotes monocyte-derived macrophages with an anti-inflammatory gene expression and functional profile A Schematic representation of the experiments. Monocytes were exposed to 10–100 nM Upadacitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Right, immunoblot analysis of pSTAT5, STAT5, pERK, ERK and pp38 by monocytes treated for 1 h to 100 nM Upadacitinib (Upa) and exposed to GM-CSF for the indicated time points. B Number of annotated genes whose expression is regulated in GM-MØ after 7d of Upadacitinib treatment (adj p < 0.05). C Volcano plot of RNAseq results showing the 100 nM Upadacitinib-induced gene expression changes in GM-MØ. D PCA analysis of GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ.Three independent donors are identified as I, II and III. E GSEA on the ranked comparison of the GM-MØ versus 10Upa-GM-MØ and GM-MØ versus 100Upa-GM-MØ transcriptomes, using the genes significantly modulated by GM-CSF (GM-MØ-specific markers) and M-CSF (M-MØ-specific markers) as data set. Normalized Enrichment Score (NES) and False Discovery Rate (FDRq) are indicated. F Relative expression of the indicated genes as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . G Production of activin A, IL-10 and LGMN by GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 8 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test; F = 92.63 for Activin A, F = 17.85 for IL-10, F = 33.39 for LGMN). H Immunoblot analysis of CD163 and FOLR2 (down) by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and monocytes differentiated with M-CSF (M-MØ). In panels A-G, vinculin or GAPDH protein levels were determined as protein loading controls and a representative experiment of two independent donors is shown. I Phagocytic activity in GM-MØ, 100Upa-GM-MØ and M-MØ . Mean ± SEM of 5 independent donors are shown (* p < 0.05, one-way ANOVA with Tukey’s post hoc test, F = 13.74). J Production of TNFα, IL-6 and IL-10 by GM-MØ and 100Upa-GM-MØ challenged with LPS for 24 h, as determined by ELISA. Mean ± SEM of 7–8 independent donors are shown (* p < 0.05, ** p < 0.01, paired t-test)
    Figure Legend Snippet: Upadacitinib promotes monocyte-derived macrophages with an anti-inflammatory gene expression and functional profile A Schematic representation of the experiments. Monocytes were exposed to 10–100 nM Upadacitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Right, immunoblot analysis of pSTAT5, STAT5, pERK, ERK and pp38 by monocytes treated for 1 h to 100 nM Upadacitinib (Upa) and exposed to GM-CSF for the indicated time points. B Number of annotated genes whose expression is regulated in GM-MØ after 7d of Upadacitinib treatment (adj p < 0.05). C Volcano plot of RNAseq results showing the 100 nM Upadacitinib-induced gene expression changes in GM-MØ. D PCA analysis of GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ.Three independent donors are identified as I, II and III. E GSEA on the ranked comparison of the GM-MØ versus 10Upa-GM-MØ and GM-MØ versus 100Upa-GM-MØ transcriptomes, using the genes significantly modulated by GM-CSF (GM-MØ-specific markers) and M-CSF (M-MØ-specific markers) as data set. Normalized Enrichment Score (NES) and False Discovery Rate (FDRq) are indicated. F Relative expression of the indicated genes as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . G Production of activin A, IL-10 and LGMN by GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 8 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test; F = 92.63 for Activin A, F = 17.85 for IL-10, F = 33.39 for LGMN). H Immunoblot analysis of CD163 and FOLR2 (down) by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and monocytes differentiated with M-CSF (M-MØ). In panels A-G, vinculin or GAPDH protein levels were determined as protein loading controls and a representative experiment of two independent donors is shown. I Phagocytic activity in GM-MØ, 100Upa-GM-MØ and M-MØ . Mean ± SEM of 5 independent donors are shown (* p < 0.05, one-way ANOVA with Tukey’s post hoc test, F = 13.74). J Production of TNFα, IL-6 and IL-10 by GM-MØ and 100Upa-GM-MØ challenged with LPS for 24 h, as determined by ELISA. Mean ± SEM of 7–8 independent donors are shown (* p < 0.05, ** p < 0.01, paired t-test)

    Techniques Used: Derivative Assay, Gene Expression, Functional Assay, Western Blot, Expressing, Comparison, RNA Sequencing, Activity Assay, Enzyme-linked Immunosorbent Assay

    JAKi increases the expression of MAFB transcription factor in macrophages A Discriminant regulon expression analysis (DoRothEA) of 100Upa-GM-MØ compared with GM-MØ. Top 25 transcription factors are shown. B GSEA of genes downregulated by siRNA MAFB and by siRNA MAF on macrophages (GSE155719) on the ranked comparison of the transcriptomes of 100Upa-GM-MØ and GM-MØ transcriptomes. NES and FDRq value are indicated. C Relative expression of MAFB and MAF as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . D Immunoblot analysis of MAFB and pGSK3S9 by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and M-MØ. E Immunoblot analysis of MAFB and CD163 along the monocyte to macrophage differentiation in the presence of 100 nM Upadacitinib (Upa). In panels (D-E), GAPDH protein levels were determined as protein loading control and a representative experiment of two ( E ) and four ( D ) independent donors is shown. F Schematic representation of the experiments: short term-Upadacitinib treatment to mature macropahges (GM-MØ). Immunoblot analysis of MAFB and pGSK3S9 ( G ) and production of activin A ( H ) by GM-MØ exposed to 10–100 nM Upadacitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( G ) a representative experiment of three independent donors is shown. In ( H ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 12.28). I Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes exposed to DMSO (−), Upadacitinib (Upa, 100 nM), STAT5 phosphorylation specific inhibitor (ST5i, 50 µM) or MEK1/2 inhibitor (UO, 2,5 µM). Right, quantification of MAFB expression. J Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes transfected with either siCNT or MAFB-specific siRNA (siMAFB) and exposed to DMSO (−) or 100 nM Upadacitinib (+). Right, quantification of MAFB expression. In panels I-J, mean ± SEM of the relative MAFB protein levels in the macrophage subtypes from four independent donors are shown (* p < 0.05, ** p < 0.01). K Relative mRNA expression of the indicated MAFB-dependent genes in siCNT GM-MØ, siMAFB GM-MØ, siCNT Upa-GM-MØ and siMAFB Upa-GM-MØ (day 2). Mean ± SEM of four independent experiments are shown (* p < 0.05; ** p < 0.01; *** p < 0.001, F = 3.7 for IL10 , F = 6.8 for CMKLR1 , F = 4.4 for LGMN , F = 56.89 for CD163 , F = 9.24 for FOLR2 , F = 8.34 for MS4A6A )
    Figure Legend Snippet: JAKi increases the expression of MAFB transcription factor in macrophages A Discriminant regulon expression analysis (DoRothEA) of 100Upa-GM-MØ compared with GM-MØ. Top 25 transcription factors are shown. B GSEA of genes downregulated by siRNA MAFB and by siRNA MAF on macrophages (GSE155719) on the ranked comparison of the transcriptomes of 100Upa-GM-MØ and GM-MØ transcriptomes. NES and FDRq value are indicated. C Relative expression of MAFB and MAF as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . D Immunoblot analysis of MAFB and pGSK3S9 by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and M-MØ. E Immunoblot analysis of MAFB and CD163 along the monocyte to macrophage differentiation in the presence of 100 nM Upadacitinib (Upa). In panels (D-E), GAPDH protein levels were determined as protein loading control and a representative experiment of two ( E ) and four ( D ) independent donors is shown. F Schematic representation of the experiments: short term-Upadacitinib treatment to mature macropahges (GM-MØ). Immunoblot analysis of MAFB and pGSK3S9 ( G ) and production of activin A ( H ) by GM-MØ exposed to 10–100 nM Upadacitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( G ) a representative experiment of three independent donors is shown. In ( H ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 12.28). I Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes exposed to DMSO (−), Upadacitinib (Upa, 100 nM), STAT5 phosphorylation specific inhibitor (ST5i, 50 µM) or MEK1/2 inhibitor (UO, 2,5 µM). Right, quantification of MAFB expression. J Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes transfected with either siCNT or MAFB-specific siRNA (siMAFB) and exposed to DMSO (−) or 100 nM Upadacitinib (+). Right, quantification of MAFB expression. In panels I-J, mean ± SEM of the relative MAFB protein levels in the macrophage subtypes from four independent donors are shown (* p < 0.05, ** p < 0.01). K Relative mRNA expression of the indicated MAFB-dependent genes in siCNT GM-MØ, siMAFB GM-MØ, siCNT Upa-GM-MØ and siMAFB Upa-GM-MØ (day 2). Mean ± SEM of four independent experiments are shown (* p < 0.05; ** p < 0.01; *** p < 0.001, F = 3.7 for IL10 , F = 6.8 for CMKLR1 , F = 4.4 for LGMN , F = 56.89 for CD163 , F = 9.24 for FOLR2 , F = 8.34 for MS4A6A )

    Techniques Used: Expressing, Comparison, RNA Sequencing, Western Blot, Control, Generated, Phospho-proteomics, Transfection

    Macrophage re-programming by other JAK inhibitors A Monocytes were exposed to 10 nM Baricitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ and 10Bari-GM-MØ. GSEA on the ranked comparison of the GM-MØ versus 10Bari-GM-MØ transcriptomes, using the genes preferentially expressed by GM-CSF (GM-MØ-specific) and M-CSF (M-MØ-specific) (GSE188278) and RA-specific clusters of synovial tissue macrophages (E-MTAB-8322) as data set. NES and FDRq value are indicated (FDRq < 0.01, dark filled circle; FDRq > 0.250, empty circle). B Production of activin A, IL-10 and LGMN by GM-MØ and 10Bari-GM-MØ. Mean ± SEM of 8–9 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test). C – D Immunoblot analysis of FOLR2 ( C ) MAFB, CD163 and pGSK3S9 ( D ) by GM-MØ, 10Bari-GM-MØ, 100Bari-GM-MØ and monocytes differentiated with M-CSF (M-MØ). Vinculin and GAPDH protein levels were determined as protein loading control. A representative experiment of two ( C ) and four ( D ) independent donors is shown. E Schematic representation of the experiments: short-term Baricitinib treatment to GM-MØ. Immunoblot analysis of MAFB and pGSK3S9 ( F ) and production of activin A ( G ) by GM-MØ exposed to 10–100 nM Baricitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( F ) a representative experiment of three independent donors is shown. In ( G ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 7.26). H Immunoblot analysis of MAFB and CD163 in GM-MØ (day 2) generated from monocytes exposed to DMSO (−) or 100 nM Tofacitinib (Tofa), Baricitinib (Bari), Upadacitinib (Upa), Peficitinib (Pefi), Filgotinib (Filgo) or Deucravacitinib (Deucra). A representative experiment of two independent donors is shown
    Figure Legend Snippet: Macrophage re-programming by other JAK inhibitors A Monocytes were exposed to 10 nM Baricitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ and 10Bari-GM-MØ. GSEA on the ranked comparison of the GM-MØ versus 10Bari-GM-MØ transcriptomes, using the genes preferentially expressed by GM-CSF (GM-MØ-specific) and M-CSF (M-MØ-specific) (GSE188278) and RA-specific clusters of synovial tissue macrophages (E-MTAB-8322) as data set. NES and FDRq value are indicated (FDRq < 0.01, dark filled circle; FDRq > 0.250, empty circle). B Production of activin A, IL-10 and LGMN by GM-MØ and 10Bari-GM-MØ. Mean ± SEM of 8–9 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test). C – D Immunoblot analysis of FOLR2 ( C ) MAFB, CD163 and pGSK3S9 ( D ) by GM-MØ, 10Bari-GM-MØ, 100Bari-GM-MØ and monocytes differentiated with M-CSF (M-MØ). Vinculin and GAPDH protein levels were determined as protein loading control. A representative experiment of two ( C ) and four ( D ) independent donors is shown. E Schematic representation of the experiments: short-term Baricitinib treatment to GM-MØ. Immunoblot analysis of MAFB and pGSK3S9 ( F ) and production of activin A ( G ) by GM-MØ exposed to 10–100 nM Baricitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( F ) a representative experiment of three independent donors is shown. In ( G ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 7.26). H Immunoblot analysis of MAFB and CD163 in GM-MØ (day 2) generated from monocytes exposed to DMSO (−) or 100 nM Tofacitinib (Tofa), Baricitinib (Bari), Upadacitinib (Upa), Peficitinib (Pefi), Filgotinib (Filgo) or Deucravacitinib (Deucra). A representative experiment of two independent donors is shown

    Techniques Used: Comparison, Western Blot, Control, Generated

    Related Articles

    Derivative Assay:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Gene Expression:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Functional Assay:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Western Blot:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Expressing:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Comparison:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    RNA Sequencing:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Activity Assay:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Control:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Generated:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Phospho-proteomics:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).

    Transfection:

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB
    Article Snippet: For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.For folate receptor beta (FOLR2), cell lysates were subjected to SDS-PAGE under non-reduced conditions.. Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).. Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).Protein loading was normalized using an antibody against GAPDH (6C5, Santa Cruz Biotechnology, 1/2000) or against human vinculin (clone VIN-11-5, Sigma-Aldrich, 1/3000).



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    Upadacitinib promotes monocyte-derived macrophages with an anti-inflammatory gene expression and functional profile A Schematic representation of the experiments. Monocytes were exposed to 10–100 nM Upadacitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Right, immunoblot analysis of pSTAT5, STAT5, pERK, ERK and pp38 by monocytes treated for 1 h to 100 nM Upadacitinib (Upa) and exposed to GM-CSF for the indicated time points. B Number of annotated genes whose expression is regulated in GM-MØ after 7d of Upadacitinib treatment (adj p < 0.05). C Volcano plot of RNAseq results showing the 100 nM Upadacitinib-induced gene expression changes in GM-MØ. D PCA analysis of GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ.Three independent donors are identified as I, II and III. E GSEA on the ranked comparison of the GM-MØ versus 10Upa-GM-MØ and GM-MØ versus 100Upa-GM-MØ transcriptomes, using the genes significantly modulated by GM-CSF (GM-MØ-specific markers) and M-CSF (M-MØ-specific markers) as data set. Normalized Enrichment Score (NES) and False Discovery Rate (FDRq) are indicated. F Relative expression of the indicated genes as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . G Production of activin A, IL-10 and LGMN by GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 8 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test; F = 92.63 for Activin A, F = 17.85 for IL-10, F = 33.39 for LGMN). H Immunoblot analysis of CD163 and FOLR2 (down) by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and monocytes differentiated with M-CSF (M-MØ). In panels A-G, vinculin or GAPDH protein levels were determined as protein loading controls and a representative experiment of two independent donors is shown. I Phagocytic activity in GM-MØ, 100Upa-GM-MØ and M-MØ . Mean ± SEM of 5 independent donors are shown (* p < 0.05, one-way ANOVA with Tukey’s post hoc test, F = 13.74). J Production of TNFα, IL-6 and IL-10 by GM-MØ and 100Upa-GM-MØ challenged with LPS for 24 h, as determined by ELISA. Mean ± SEM of 7–8 independent donors are shown (* p < 0.05, ** p < 0.01, paired t-test)

    Journal: Cellular and Molecular Life Sciences

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB

    doi: 10.1007/s00018-024-05196-1

    Figure Lengend Snippet: Upadacitinib promotes monocyte-derived macrophages with an anti-inflammatory gene expression and functional profile A Schematic representation of the experiments. Monocytes were exposed to 10–100 nM Upadacitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Right, immunoblot analysis of pSTAT5, STAT5, pERK, ERK and pp38 by monocytes treated for 1 h to 100 nM Upadacitinib (Upa) and exposed to GM-CSF for the indicated time points. B Number of annotated genes whose expression is regulated in GM-MØ after 7d of Upadacitinib treatment (adj p < 0.05). C Volcano plot of RNAseq results showing the 100 nM Upadacitinib-induced gene expression changes in GM-MØ. D PCA analysis of GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ.Three independent donors are identified as I, II and III. E GSEA on the ranked comparison of the GM-MØ versus 10Upa-GM-MØ and GM-MØ versus 100Upa-GM-MØ transcriptomes, using the genes significantly modulated by GM-CSF (GM-MØ-specific markers) and M-CSF (M-MØ-specific markers) as data set. Normalized Enrichment Score (NES) and False Discovery Rate (FDRq) are indicated. F Relative expression of the indicated genes as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . G Production of activin A, IL-10 and LGMN by GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 8 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test; F = 92.63 for Activin A, F = 17.85 for IL-10, F = 33.39 for LGMN). H Immunoblot analysis of CD163 and FOLR2 (down) by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and monocytes differentiated with M-CSF (M-MØ). In panels A-G, vinculin or GAPDH protein levels were determined as protein loading controls and a representative experiment of two independent donors is shown. I Phagocytic activity in GM-MØ, 100Upa-GM-MØ and M-MØ . Mean ± SEM of 5 independent donors are shown (* p < 0.05, one-way ANOVA with Tukey’s post hoc test, F = 13.74). J Production of TNFα, IL-6 and IL-10 by GM-MØ and 100Upa-GM-MØ challenged with LPS for 24 h, as determined by ELISA. Mean ± SEM of 7–8 independent donors are shown (* p < 0.05, ** p < 0.01, paired t-test)

    Article Snippet: Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).

    Techniques: Derivative Assay, Gene Expression, Functional Assay, Western Blot, Expressing, Comparison, RNA Sequencing, Activity Assay, Enzyme-linked Immunosorbent Assay

    JAKi increases the expression of MAFB transcription factor in macrophages A Discriminant regulon expression analysis (DoRothEA) of 100Upa-GM-MØ compared with GM-MØ. Top 25 transcription factors are shown. B GSEA of genes downregulated by siRNA MAFB and by siRNA MAF on macrophages (GSE155719) on the ranked comparison of the transcriptomes of 100Upa-GM-MØ and GM-MØ transcriptomes. NES and FDRq value are indicated. C Relative expression of MAFB and MAF as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . D Immunoblot analysis of MAFB and pGSK3S9 by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and M-MØ. E Immunoblot analysis of MAFB and CD163 along the monocyte to macrophage differentiation in the presence of 100 nM Upadacitinib (Upa). In panels (D-E), GAPDH protein levels were determined as protein loading control and a representative experiment of two ( E ) and four ( D ) independent donors is shown. F Schematic representation of the experiments: short term-Upadacitinib treatment to mature macropahges (GM-MØ). Immunoblot analysis of MAFB and pGSK3S9 ( G ) and production of activin A ( H ) by GM-MØ exposed to 10–100 nM Upadacitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( G ) a representative experiment of three independent donors is shown. In ( H ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 12.28). I Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes exposed to DMSO (−), Upadacitinib (Upa, 100 nM), STAT5 phosphorylation specific inhibitor (ST5i, 50 µM) or MEK1/2 inhibitor (UO, 2,5 µM). Right, quantification of MAFB expression. J Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes transfected with either siCNT or MAFB-specific siRNA (siMAFB) and exposed to DMSO (−) or 100 nM Upadacitinib (+). Right, quantification of MAFB expression. In panels I-J, mean ± SEM of the relative MAFB protein levels in the macrophage subtypes from four independent donors are shown (* p < 0.05, ** p < 0.01). K Relative mRNA expression of the indicated MAFB-dependent genes in siCNT GM-MØ, siMAFB GM-MØ, siCNT Upa-GM-MØ and siMAFB Upa-GM-MØ (day 2). Mean ± SEM of four independent experiments are shown (* p < 0.05; ** p < 0.01; *** p < 0.001, F = 3.7 for IL10 , F = 6.8 for CMKLR1 , F = 4.4 for LGMN , F = 56.89 for CD163 , F = 9.24 for FOLR2 , F = 8.34 for MS4A6A )

    Journal: Cellular and Molecular Life Sciences

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB

    doi: 10.1007/s00018-024-05196-1

    Figure Lengend Snippet: JAKi increases the expression of MAFB transcription factor in macrophages A Discriminant regulon expression analysis (DoRothEA) of 100Upa-GM-MØ compared with GM-MØ. Top 25 transcription factors are shown. B GSEA of genes downregulated by siRNA MAFB and by siRNA MAF on macrophages (GSE155719) on the ranked comparison of the transcriptomes of 100Upa-GM-MØ and GM-MØ transcriptomes. NES and FDRq value are indicated. C Relative expression of MAFB and MAF as determined by RNA-sequencing on GM-MØ, 10Upa-GM-MØ and 100Upa-GM-MØ. Mean ± SEM of 4 independent donors are shown, with the indication of the P adj . D Immunoblot analysis of MAFB and pGSK3S9 by GM-MØ, 10Upa-GM-MØ, 100Upa-GM-MØ and M-MØ. E Immunoblot analysis of MAFB and CD163 along the monocyte to macrophage differentiation in the presence of 100 nM Upadacitinib (Upa). In panels (D-E), GAPDH protein levels were determined as protein loading control and a representative experiment of two ( E ) and four ( D ) independent donors is shown. F Schematic representation of the experiments: short term-Upadacitinib treatment to mature macropahges (GM-MØ). Immunoblot analysis of MAFB and pGSK3S9 ( G ) and production of activin A ( H ) by GM-MØ exposed to 10–100 nM Upadacitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( G ) a representative experiment of three independent donors is shown. In ( H ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 12.28). I Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes exposed to DMSO (−), Upadacitinib (Upa, 100 nM), STAT5 phosphorylation specific inhibitor (ST5i, 50 µM) or MEK1/2 inhibitor (UO, 2,5 µM). Right, quantification of MAFB expression. J Immunoblot analysis of MAFB in two independent preparations of differentiating GM-MØ (day 2) generated from monocytes transfected with either siCNT or MAFB-specific siRNA (siMAFB) and exposed to DMSO (−) or 100 nM Upadacitinib (+). Right, quantification of MAFB expression. In panels I-J, mean ± SEM of the relative MAFB protein levels in the macrophage subtypes from four independent donors are shown (* p < 0.05, ** p < 0.01). K Relative mRNA expression of the indicated MAFB-dependent genes in siCNT GM-MØ, siMAFB GM-MØ, siCNT Upa-GM-MØ and siMAFB Upa-GM-MØ (day 2). Mean ± SEM of four independent experiments are shown (* p < 0.05; ** p < 0.01; *** p < 0.001, F = 3.7 for IL10 , F = 6.8 for CMKLR1 , F = 4.4 for LGMN , F = 56.89 for CD163 , F = 9.24 for FOLR2 , F = 8.34 for MS4A6A )

    Article Snippet: Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).

    Techniques: Expressing, Comparison, RNA Sequencing, Western Blot, Control, Generated, Phospho-proteomics, Transfection

    Macrophage re-programming by other JAK inhibitors A Monocytes were exposed to 10 nM Baricitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ and 10Bari-GM-MØ. GSEA on the ranked comparison of the GM-MØ versus 10Bari-GM-MØ transcriptomes, using the genes preferentially expressed by GM-CSF (GM-MØ-specific) and M-CSF (M-MØ-specific) (GSE188278) and RA-specific clusters of synovial tissue macrophages (E-MTAB-8322) as data set. NES and FDRq value are indicated (FDRq < 0.01, dark filled circle; FDRq > 0.250, empty circle). B Production of activin A, IL-10 and LGMN by GM-MØ and 10Bari-GM-MØ. Mean ± SEM of 8–9 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test). C – D Immunoblot analysis of FOLR2 ( C ) MAFB, CD163 and pGSK3S9 ( D ) by GM-MØ, 10Bari-GM-MØ, 100Bari-GM-MØ and monocytes differentiated with M-CSF (M-MØ). Vinculin and GAPDH protein levels were determined as protein loading control. A representative experiment of two ( C ) and four ( D ) independent donors is shown. E Schematic representation of the experiments: short-term Baricitinib treatment to GM-MØ. Immunoblot analysis of MAFB and pGSK3S9 ( F ) and production of activin A ( G ) by GM-MØ exposed to 10–100 nM Baricitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( F ) a representative experiment of three independent donors is shown. In ( G ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 7.26). H Immunoblot analysis of MAFB and CD163 in GM-MØ (day 2) generated from monocytes exposed to DMSO (−) or 100 nM Tofacitinib (Tofa), Baricitinib (Bari), Upadacitinib (Upa), Peficitinib (Pefi), Filgotinib (Filgo) or Deucravacitinib (Deucra). A representative experiment of two independent donors is shown

    Journal: Cellular and Molecular Life Sciences

    Article Title: Macrophage re-programming by JAK inhibitors relies on MAFB

    doi: 10.1007/s00018-024-05196-1

    Figure Lengend Snippet: Macrophage re-programming by other JAK inhibitors A Monocytes were exposed to 10 nM Baricitinib daily during macrophage differentiation process with GM-CSF and the RNA levels were determined at day 7 on GM-MØ and 10Bari-GM-MØ. GSEA on the ranked comparison of the GM-MØ versus 10Bari-GM-MØ transcriptomes, using the genes preferentially expressed by GM-CSF (GM-MØ-specific) and M-CSF (M-MØ-specific) (GSE188278) and RA-specific clusters of synovial tissue macrophages (E-MTAB-8322) as data set. NES and FDRq value are indicated (FDRq < 0.01, dark filled circle; FDRq > 0.250, empty circle). B Production of activin A, IL-10 and LGMN by GM-MØ and 10Bari-GM-MØ. Mean ± SEM of 8–9 independent donors are shown (* p < 0.05, ** p < 0.01, one-way ANOVA with Tukey´s post hoc test). C – D Immunoblot analysis of FOLR2 ( C ) MAFB, CD163 and pGSK3S9 ( D ) by GM-MØ, 10Bari-GM-MØ, 100Bari-GM-MØ and monocytes differentiated with M-CSF (M-MØ). Vinculin and GAPDH protein levels were determined as protein loading control. A representative experiment of two ( C ) and four ( D ) independent donors is shown. E Schematic representation of the experiments: short-term Baricitinib treatment to GM-MØ. Immunoblot analysis of MAFB and pGSK3S9 ( F ) and production of activin A ( G ) by GM-MØ exposed to 10–100 nM Baricitinib for the last 48 h. GAPDH protein levels were determined as protein loading control. In ( F ) a representative experiment of three independent donors is shown. In ( G ) mean ± SEM of 5 independent donors are shown (* p < 0.05, F = 7.26). H Immunoblot analysis of MAFB and CD163 in GM-MØ (day 2) generated from monocytes exposed to DMSO (−) or 100 nM Tofacitinib (Tofa), Baricitinib (Bari), Upadacitinib (Upa), Peficitinib (Pefi), Filgotinib (Filgo) or Deucravacitinib (Deucra). A representative experiment of two independent donors is shown

    Article Snippet: Protein detection was carried out using rabbit antibodies against pp38 and pERK (clones D3F9 and D13.14.4E, Cell Signaling, 1/1000), MAFB (HPA005653, Santa Cruz, 1/1000), pGSK3β (clone D85E12, Cell Signaling, 1/1000) and mouse monoclonal antibody against human CD163 (clone EDHu-1, Bio-Rad, 1/1000), pSTAT5 (clone 8-5-2, Millipore, 1/1000), FOLR2 (FRβ, kindly provided by Dr. Takami Matsuyama [ ], dilution 1/800).

    Techniques: Comparison, Western Blot, Control, Generated

    Immunofluorescence staining of inflammatory cells in pediatric kidneys. A CD68 + macrophages. B CD163 + cells. C Merge of CD68 + and CD163 + cells representing M2c-like macrophages. D CD68 + macrophages. E CD 206 + cells. F Merge CD68 + and CD206 + cells, representing M2a-like macrophages. G Renal T lymphocytes (CD3 + cells). H Renal B lymphocytes (CD20 + cells). I B and T Lymphocyte overlap (CD3 + and CD20 + cells. Scale bar represents 50 µm

    Journal: Arthritis Research & Therapy

    Article Title: Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

    doi: 10.1186/s13075-024-03281-1

    Figure Lengend Snippet: Immunofluorescence staining of inflammatory cells in pediatric kidneys. A CD68 + macrophages. B CD163 + cells. C Merge of CD68 + and CD163 + cells representing M2c-like macrophages. D CD68 + macrophages. E CD 206 + cells. F Merge CD68 + and CD206 + cells, representing M2a-like macrophages. G Renal T lymphocytes (CD3 + cells). H Renal B lymphocytes (CD20 + cells). I B and T Lymphocyte overlap (CD3 + and CD20 + cells. Scale bar represents 50 µm

    Article Snippet: After blocking with normal goat serum and 1% blotto sections were incubated overnight at 4 °C using the following antibodies diluted in 1% BSA in 50 mM Tris(hydroxymethyl) aminomethan pH 7.6: iNOS, a rabbit polyclonal antibody against human iNOS (Abcam plc, Cambridge, UK); CD68, a mouse monoclonal IgG3 antibody against human CD68 (Dako Deutschland GmbH, Hamburg, Germany); CD163, a mouse monoclonal IgG1 antibody against human CD163 (Novocastra, Leica Biosystems Newcastle Ltd; Newcastle, UK); CD206, a mouse monoclonal IgG1 antibody against human CD206 (Abnova, Jhongli City, Taiwan); CD3, a monoclonal rat antibody against human CD3 (Bio-Rad AbD Serotec GmbH, Puchheim, Germany); CD20, a monoclonal mouse IgG2a antibody against human CD20 and MPO, a polyclonal rabbit antibody against myeloperoxidase (Abcam plc, Cambridge, UK).

    Techniques: Immunofluorescence, Staining

    Distribution of macrophage subsets in different ISN/RPS classes. A CD68 + macrophages in pediatric LN patient groups representing the total macrophages. B CD68 + CD206 − cells (M1-like macrophages) in pediatric LN patients. C CD68 + CD163 − cells (M1-like macrophages). D CD68 + as total macrophages in adult LN patients with a significant difference between groups II, V, and IV. E CD68 + CD206 − cells (M1-like macrophages) with a significant difference between groups IV and V. F CD 68 + CD163 − cells ((M1-like macrophages) ( E and F in adult LN patients). G – I Showing results for pediatric LN patients, G CD68 + CD206 + cells (M2a-like macrophages), H CD68 + CD163 + cells (M2c-like macrophages), I Ration of CD68 + CD206 − vs CD68 + CD206 + cells (M1-like macrophages and M2a-like macrophages) with a significant difference between groups II and IV. J – L representing adult LN results. J CD68 + CD206 + cells (M2a-like macrophages), K CD68 + CD163 + cells (M2c-like macrophages), with a significant difference between groups II, V, and IV. L Ration of CD68 + CD206 − vs CD68 + CD206 + cells (M1-like macrophages and M2a-like macrophages). (* p < 0.05; ** p < 0.01)

    Journal: Arthritis Research & Therapy

    Article Title: Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

    doi: 10.1186/s13075-024-03281-1

    Figure Lengend Snippet: Distribution of macrophage subsets in different ISN/RPS classes. A CD68 + macrophages in pediatric LN patient groups representing the total macrophages. B CD68 + CD206 − cells (M1-like macrophages) in pediatric LN patients. C CD68 + CD163 − cells (M1-like macrophages). D CD68 + as total macrophages in adult LN patients with a significant difference between groups II, V, and IV. E CD68 + CD206 − cells (M1-like macrophages) with a significant difference between groups IV and V. F CD 68 + CD163 − cells ((M1-like macrophages) ( E and F in adult LN patients). G – I Showing results for pediatric LN patients, G CD68 + CD206 + cells (M2a-like macrophages), H CD68 + CD163 + cells (M2c-like macrophages), I Ration of CD68 + CD206 − vs CD68 + CD206 + cells (M1-like macrophages and M2a-like macrophages) with a significant difference between groups II and IV. J – L representing adult LN results. J CD68 + CD206 + cells (M2a-like macrophages), K CD68 + CD163 + cells (M2c-like macrophages), with a significant difference between groups II, V, and IV. L Ration of CD68 + CD206 − vs CD68 + CD206 + cells (M1-like macrophages and M2a-like macrophages). (* p < 0.05; ** p < 0.01)

    Article Snippet: After blocking with normal goat serum and 1% blotto sections were incubated overnight at 4 °C using the following antibodies diluted in 1% BSA in 50 mM Tris(hydroxymethyl) aminomethan pH 7.6: iNOS, a rabbit polyclonal antibody against human iNOS (Abcam plc, Cambridge, UK); CD68, a mouse monoclonal IgG3 antibody against human CD68 (Dako Deutschland GmbH, Hamburg, Germany); CD163, a mouse monoclonal IgG1 antibody against human CD163 (Novocastra, Leica Biosystems Newcastle Ltd; Newcastle, UK); CD206, a mouse monoclonal IgG1 antibody against human CD206 (Abnova, Jhongli City, Taiwan); CD3, a monoclonal rat antibody against human CD3 (Bio-Rad AbD Serotec GmbH, Puchheim, Germany); CD20, a monoclonal mouse IgG2a antibody against human CD20 and MPO, a polyclonal rabbit antibody against myeloperoxidase (Abcam plc, Cambridge, UK).

    Techniques:

    Distribution of macrophage-subtypes in pediatric LN ISN/RPN class IV in glomeruli with and without (w/o) crescents. A Representative multiple immunofluorescence staining for CD68 + and CD163 + cells using biopsies from patients with ISN/RPN class IV. B Multiple immunofluorescence staining for CD68 + , CD206 + CD20 + , and CD3 + cells using the same glomerulus imaged in ( A ) in another section. C Pie chart showing the distribution of different inflammatory cells in glomeruli from pediatric LN biopsies with ISN/RPN class IV. D MPO + cells /glomerular cross-section (GCS). E CD3 + T-cells / GCS. F Glomerular CD20 + B-cells. G Glomerular total CD68 + macrophages per GCS (first antibody panel). H Glomerular CD68 + CD206 − M1-like macrophages/ GCS. I Glomerular CD68 + CD206 + M2a-like macrophages. J Ratio of glomerular CD68 + CD206 − /CD68 + CD206 + macrophages. K glomerular total CD68 + macrophages per GCS (second antibody panel). H Glomerular CD68 + CD163 − M1-like macrophages/ GCS. I Glomerular CD68 + CD163 + M2c-like macrophages. J Ratio of glomerular CD68 + CD163 − /CD68 + CD163 + macrophages. (* p < 0,05). Scale bar represents 50 µm

    Journal: Arthritis Research & Therapy

    Article Title: Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

    doi: 10.1186/s13075-024-03281-1

    Figure Lengend Snippet: Distribution of macrophage-subtypes in pediatric LN ISN/RPN class IV in glomeruli with and without (w/o) crescents. A Representative multiple immunofluorescence staining for CD68 + and CD163 + cells using biopsies from patients with ISN/RPN class IV. B Multiple immunofluorescence staining for CD68 + , CD206 + CD20 + , and CD3 + cells using the same glomerulus imaged in ( A ) in another section. C Pie chart showing the distribution of different inflammatory cells in glomeruli from pediatric LN biopsies with ISN/RPN class IV. D MPO + cells /glomerular cross-section (GCS). E CD3 + T-cells / GCS. F Glomerular CD20 + B-cells. G Glomerular total CD68 + macrophages per GCS (first antibody panel). H Glomerular CD68 + CD206 − M1-like macrophages/ GCS. I Glomerular CD68 + CD206 + M2a-like macrophages. J Ratio of glomerular CD68 + CD206 − /CD68 + CD206 + macrophages. K glomerular total CD68 + macrophages per GCS (second antibody panel). H Glomerular CD68 + CD163 − M1-like macrophages/ GCS. I Glomerular CD68 + CD163 + M2c-like macrophages. J Ratio of glomerular CD68 + CD163 − /CD68 + CD163 + macrophages. (* p < 0,05). Scale bar represents 50 µm

    Article Snippet: After blocking with normal goat serum and 1% blotto sections were incubated overnight at 4 °C using the following antibodies diluted in 1% BSA in 50 mM Tris(hydroxymethyl) aminomethan pH 7.6: iNOS, a rabbit polyclonal antibody against human iNOS (Abcam plc, Cambridge, UK); CD68, a mouse monoclonal IgG3 antibody against human CD68 (Dako Deutschland GmbH, Hamburg, Germany); CD163, a mouse monoclonal IgG1 antibody against human CD163 (Novocastra, Leica Biosystems Newcastle Ltd; Newcastle, UK); CD206, a mouse monoclonal IgG1 antibody against human CD206 (Abnova, Jhongli City, Taiwan); CD3, a monoclonal rat antibody against human CD3 (Bio-Rad AbD Serotec GmbH, Puchheim, Germany); CD20, a monoclonal mouse IgG2a antibody against human CD20 and MPO, a polyclonal rabbit antibody against myeloperoxidase (Abcam plc, Cambridge, UK).

    Techniques: Immunofluorescence, Staining

    Distribution of macrophage-subtypes in pediatric LN and a pediatric control group consisting of HUS, MPGN, PI-GN and PAUCI patients. A CD68 + total macrophages in the inflammatory kidney in pediatric patients with significantly higher in PAUCI patients compared to the LN and control group. B CD68 + CD206 − M1-like macrophages significantly higher in pediatric PAUCI patients compared to all other study groups. C CD68 + CD163 − M1-like macrophages significantly higher in PAUCI than in LN patients. D CD68 + CD206 + M2a-like macrophages, significantly higher in PAUCI than in LN patients. E CD68 + CD163 + M2c-like macrophages, significantly higher in HUS pediatric patients compared to MPGN, LN patients and our pediatric control group. F Ratio of CD68 + CD206 − and CD68 + CD206 + (M1-like and M2a-like macrophages) in pediatric patients. (* p < 0.05; ** p < 0.01)

    Journal: Arthritis Research & Therapy

    Article Title: Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

    doi: 10.1186/s13075-024-03281-1

    Figure Lengend Snippet: Distribution of macrophage-subtypes in pediatric LN and a pediatric control group consisting of HUS, MPGN, PI-GN and PAUCI patients. A CD68 + total macrophages in the inflammatory kidney in pediatric patients with significantly higher in PAUCI patients compared to the LN and control group. B CD68 + CD206 − M1-like macrophages significantly higher in pediatric PAUCI patients compared to all other study groups. C CD68 + CD163 − M1-like macrophages significantly higher in PAUCI than in LN patients. D CD68 + CD206 + M2a-like macrophages, significantly higher in PAUCI than in LN patients. E CD68 + CD163 + M2c-like macrophages, significantly higher in HUS pediatric patients compared to MPGN, LN patients and our pediatric control group. F Ratio of CD68 + CD206 − and CD68 + CD206 + (M1-like and M2a-like macrophages) in pediatric patients. (* p < 0.05; ** p < 0.01)

    Article Snippet: After blocking with normal goat serum and 1% blotto sections were incubated overnight at 4 °C using the following antibodies diluted in 1% BSA in 50 mM Tris(hydroxymethyl) aminomethan pH 7.6: iNOS, a rabbit polyclonal antibody against human iNOS (Abcam plc, Cambridge, UK); CD68, a mouse monoclonal IgG3 antibody against human CD68 (Dako Deutschland GmbH, Hamburg, Germany); CD163, a mouse monoclonal IgG1 antibody against human CD163 (Novocastra, Leica Biosystems Newcastle Ltd; Newcastle, UK); CD206, a mouse monoclonal IgG1 antibody against human CD206 (Abnova, Jhongli City, Taiwan); CD3, a monoclonal rat antibody against human CD3 (Bio-Rad AbD Serotec GmbH, Puchheim, Germany); CD20, a monoclonal mouse IgG2a antibody against human CD20 and MPO, a polyclonal rabbit antibody against myeloperoxidase (Abcam plc, Cambridge, UK).

    Techniques:

    Clustering analysis of the abundance of inflammatory cells in different inflammatory kidney diseases in pediatric patients. Heatmap of the numbers of CD68 + CD163 + , CD68 + CD206 + , CD68 + CD163 − , CD68 + CD206 − , CD3 + , CD20 + , and MPO + cells in the analyzed patients (red: high number; blue: low number). The color-coded bar corresponding to the different rows (patients) indicates the diagnosis

    Journal: Arthritis Research & Therapy

    Article Title: Macrophage subpopulations in pediatric patients with lupus nephritis and other inflammatory diseases affecting the kidney

    doi: 10.1186/s13075-024-03281-1

    Figure Lengend Snippet: Clustering analysis of the abundance of inflammatory cells in different inflammatory kidney diseases in pediatric patients. Heatmap of the numbers of CD68 + CD163 + , CD68 + CD206 + , CD68 + CD163 − , CD68 + CD206 − , CD3 + , CD20 + , and MPO + cells in the analyzed patients (red: high number; blue: low number). The color-coded bar corresponding to the different rows (patients) indicates the diagnosis

    Article Snippet: After blocking with normal goat serum and 1% blotto sections were incubated overnight at 4 °C using the following antibodies diluted in 1% BSA in 50 mM Tris(hydroxymethyl) aminomethan pH 7.6: iNOS, a rabbit polyclonal antibody against human iNOS (Abcam plc, Cambridge, UK); CD68, a mouse monoclonal IgG3 antibody against human CD68 (Dako Deutschland GmbH, Hamburg, Germany); CD163, a mouse monoclonal IgG1 antibody against human CD163 (Novocastra, Leica Biosystems Newcastle Ltd; Newcastle, UK); CD206, a mouse monoclonal IgG1 antibody against human CD206 (Abnova, Jhongli City, Taiwan); CD3, a monoclonal rat antibody against human CD3 (Bio-Rad AbD Serotec GmbH, Puchheim, Germany); CD20, a monoclonal mouse IgG2a antibody against human CD20 and MPO, a polyclonal rabbit antibody against myeloperoxidase (Abcam plc, Cambridge, UK).

    Techniques:

    CD163, ciliary body in horse No. 3. CD163-positive cells are located in the neighborhood of the ciliary epithelium (arrowheads). CE: ciliary epithelium. S: stroma. Bar=100 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Distribution of CD163-positive cell and MHC class II-positive cell in the normal equine uveal tract

    doi: 10.1292/jvms.15-0406

    Figure Lengend Snippet: CD163, ciliary body in horse No. 3. CD163-positive cells are located in the neighborhood of the ciliary epithelium (arrowheads). CE: ciliary epithelium. S: stroma. Bar=100 µ m.

    Article Snippet: Immunohistochemical examinations : Indirect immunofluorescence studies were performed using primary mouse monoclonal antibodies against human CD163 (AM-3K; Trans Genic Inc., Tokyo, Japan; diluted 1:50) and MHC II (HLA-DR; TAL.1B5; Dako, Glostrup, Denmark; diluted 1:50) and a rabbit polyclonal antibody against human CD20 (Thermo Fisher Scientific Inc., Waltham, MA, U.S.A.; diluted 1:200).

    Techniques:

    CD163, ciliary body in horse No. 3. Fusiform and elongate CD163-positive cells lie closely beneath and along the basal side of the pigmented ciliary epithelium (arrowheads). NE: nonpigmented epithelium. PE: pigmented epithelium. S: stroma. Bar=50 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Distribution of CD163-positive cell and MHC class II-positive cell in the normal equine uveal tract

    doi: 10.1292/jvms.15-0406

    Figure Lengend Snippet: CD163, ciliary body in horse No. 3. Fusiform and elongate CD163-positive cells lie closely beneath and along the basal side of the pigmented ciliary epithelium (arrowheads). NE: nonpigmented epithelium. PE: pigmented epithelium. S: stroma. Bar=50 µ m.

    Article Snippet: Immunohistochemical examinations : Indirect immunofluorescence studies were performed using primary mouse monoclonal antibodies against human CD163 (AM-3K; Trans Genic Inc., Tokyo, Japan; diluted 1:50) and MHC II (HLA-DR; TAL.1B5; Dako, Glostrup, Denmark; diluted 1:50) and a rabbit polyclonal antibody against human CD20 (Thermo Fisher Scientific Inc., Waltham, MA, U.S.A.; diluted 1:200).

    Techniques:

    CD163, iris in horse No.1. CD163-positive cells are scattered throughout the iridal stroma. Bar=50 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Distribution of CD163-positive cell and MHC class II-positive cell in the normal equine uveal tract

    doi: 10.1292/jvms.15-0406

    Figure Lengend Snippet: CD163, iris in horse No.1. CD163-positive cells are scattered throughout the iridal stroma. Bar=50 µ m.

    Article Snippet: Immunohistochemical examinations : Indirect immunofluorescence studies were performed using primary mouse monoclonal antibodies against human CD163 (AM-3K; Trans Genic Inc., Tokyo, Japan; diluted 1:50) and MHC II (HLA-DR; TAL.1B5; Dako, Glostrup, Denmark; diluted 1:50) and a rabbit polyclonal antibody against human CD20 (Thermo Fisher Scientific Inc., Waltham, MA, U.S.A.; diluted 1:200).

    Techniques:

    Quantitative analysis of CD163+ cells located on the ciliary body, iris and choroid. There were significant differences between the ciliary body and iris ( P <0.001) and between the ciliary body and choroid ( P =0.0021), respectively. ˟Standard error of the mean. * Statistically significant difference ( P <0.01).

    Journal: The Journal of Veterinary Medical Science

    Article Title: Distribution of CD163-positive cell and MHC class II-positive cell in the normal equine uveal tract

    doi: 10.1292/jvms.15-0406

    Figure Lengend Snippet: Quantitative analysis of CD163+ cells located on the ciliary body, iris and choroid. There were significant differences between the ciliary body and iris ( P <0.001) and between the ciliary body and choroid ( P =0.0021), respectively. ˟Standard error of the mean. * Statistically significant difference ( P <0.01).

    Article Snippet: Immunohistochemical examinations : Indirect immunofluorescence studies were performed using primary mouse monoclonal antibodies against human CD163 (AM-3K; Trans Genic Inc., Tokyo, Japan; diluted 1:50) and MHC II (HLA-DR; TAL.1B5; Dako, Glostrup, Denmark; diluted 1:50) and a rabbit polyclonal antibody against human CD20 (Thermo Fisher Scientific Inc., Waltham, MA, U.S.A.; diluted 1:200).

    Techniques:

    Expression of macrophage-specific antigen CD163 in colorectal cancer. Immunostaining shows (a) normal colon mucosa with no CD163 expression in epithelial cells. (b) Positive CD163 expression in cancer cells is manifested as cytoplasmatic and membrane staining. (c) Tumor with negative CD163 expression in cancer cells. Non-neoplastic cells staining for CD163 demonstrate tumor-associated macrophages

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: Expression of macrophage-specific antigen CD163 in colorectal cancer. Immunostaining shows (a) normal colon mucosa with no CD163 expression in epithelial cells. (b) Positive CD163 expression in cancer cells is manifested as cytoplasmatic and membrane staining. (c) Tumor with negative CD163 expression in cancer cells. Non-neoplastic cells staining for CD163 demonstrate tumor-associated macrophages

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Expressing, Immunostaining, Membrane, Staining

    Infiltration of tumor-associated macrophages (TAM) in colorectal cancer. TAM are stained with macrophage-specific antigen CD163. The sections show immunostaining representing different grades of macrophage infiltration; (a) no/low, (b) moderate, (c) high and (d) massive

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: Infiltration of tumor-associated macrophages (TAM) in colorectal cancer. TAM are stained with macrophage-specific antigen CD163. The sections show immunostaining representing different grades of macrophage infiltration; (a) no/low, (b) moderate, (c) high and (d) massive

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Staining, Immunostaining

    CD163 expression and macrophage infiltration in relation to clinical and biologic data

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: CD163 expression and macrophage infiltration in relation to clinical and biologic data

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Expressing, Marker

    Macrophage infiltration in colorectal tumor stroma in relation to CD163 expression in cancer cells. CD163 expression was significantly more frequent in tumors with high macrophage infiltration (P = 0.018)

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: Macrophage infiltration in colorectal tumor stroma in relation to CD163 expression in cancer cells. CD163 expression was significantly more frequent in tumors with high macrophage infiltration (P = 0.018)

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Expressing

    Kaplan-Meier survival curves for 75 patients with colorectal cancer. Patients with positive CD163 expression (a) and high macrophage infiltration (b) have lower survival time. The survival analysis is based on disease (CRC) specific mortality, and overall comparison is estimated according to Lon Rank (Mantel-Cox) analysis

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: Kaplan-Meier survival curves for 75 patients with colorectal cancer. Patients with positive CD163 expression (a) and high macrophage infiltration (b) have lower survival time. The survival analysis is based on disease (CRC) specific mortality, and overall comparison is estimated according to Lon Rank (Mantel-Cox) analysis

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Expressing, Comparison

    Multivariate analysis of mortality in colorectal cancer in relation to CD163 expression, macrophage infiltration and tumor stage estimated according to Cox proportional hazard analysis

    Journal: Cancer Microenvironment

    Article Title: Macrophage Infiltration in Tumor Stroma is Related to Tumor Cell Expression of CD163 in Colorectal Cancer

    doi: 10.1007/s12307-014-0145-7

    Figure Lengend Snippet: Multivariate analysis of mortality in colorectal cancer in relation to CD163 expression, macrophage infiltration and tumor stage estimated according to Cox proportional hazard analysis

    Article Snippet: Antibodies and Immune-Staining of Tissue Sections Mouse anti-human monoclonal antibody (IgG1) against CD163 (clone 10D6 from Novocastra, England) was used as a macrophage marker.

    Techniques: Expressing