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Validation of CXCL9 + and SPP1 + macrophage expression and polarization characteristics in lung cancer tissues. (A) Representative mIF staining for CXCL9+ TAMs and SPP1+ TAMs in lung tissue sections from the normal and LUAD tumor groups. DAPI (blue), <t>CD68</t> (red), CXCL9 (yellow), SPP1 (green) are shown, along with individual and merged channels. (n = 3 per group). Scale bar, 20 μm. (B) Representative mIF staining of CXCL9+ TAMs (yellow) and SPP1+ TAMs (green) in lung tissue sections from the normal and LUAD tumor groups (n = 3 per group). Scale bar, 20 μm. (C) qRT-PCR validation of macrophage polarization. The M1 group showed high expression of Cxcl9 and iNOS, while the M2 group showed high expression of Spp1 and Arg1. Asterisks indicate statistically significant differences (*p < 0.05).
Mouse Anti Human Cd68 Monoclonal Antibody, supplied by Servicebio Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
Mouse Anti Human Cd68, supplied by Bio-Rad, 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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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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( A ) Representative immunohistochemistry staining showing clear <t>CD68</t> expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.
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Image Search Results


Validation of CXCL9 + and SPP1 + macrophage expression and polarization characteristics in lung cancer tissues. (A) Representative mIF staining for CXCL9+ TAMs and SPP1+ TAMs in lung tissue sections from the normal and LUAD tumor groups. DAPI (blue), CD68 (red), CXCL9 (yellow), SPP1 (green) are shown, along with individual and merged channels. (n = 3 per group). Scale bar, 20 μm. (B) Representative mIF staining of CXCL9+ TAMs (yellow) and SPP1+ TAMs (green) in lung tissue sections from the normal and LUAD tumor groups (n = 3 per group). Scale bar, 20 μm. (C) qRT-PCR validation of macrophage polarization. The M1 group showed high expression of Cxcl9 and iNOS, while the M2 group showed high expression of Spp1 and Arg1. Asterisks indicate statistically significant differences (*p < 0.05).

Journal: Frontiers in Immunology

Article Title: The CXCL9/SPP1 polarity axis in tumor-associated macrophages: immunoregulatory and prognostic significance in non-small cell lung cancer

doi: 10.3389/fimmu.2026.1763652

Figure Lengend Snippet: Validation of CXCL9 + and SPP1 + macrophage expression and polarization characteristics in lung cancer tissues. (A) Representative mIF staining for CXCL9+ TAMs and SPP1+ TAMs in lung tissue sections from the normal and LUAD tumor groups. DAPI (blue), CD68 (red), CXCL9 (yellow), SPP1 (green) are shown, along with individual and merged channels. (n = 3 per group). Scale bar, 20 μm. (B) Representative mIF staining of CXCL9+ TAMs (yellow) and SPP1+ TAMs (green) in lung tissue sections from the normal and LUAD tumor groups (n = 3 per group). Scale bar, 20 μm. (C) qRT-PCR validation of macrophage polarization. The M1 group showed high expression of Cxcl9 and iNOS, while the M2 group showed high expression of Spp1 and Arg1. Asterisks indicate statistically significant differences (*p < 0.05).

Article Snippet: Following another stripping step, the third round was performed using mouse anti-human CD68 monoclonal antibody (1:50000, Servicebio, Cat# GB153150 ) with HRP-conjugated goat anti-mouse secondary antibody (ready-to-use, Servicebio, Cat# G1301), and signal was developed using iF555-Tyramide (1:500, Servicebio, Cat# G1233).

Techniques: Biomarker Discovery, Expressing, Staining, Quantitative RT-PCR

( A ) Representative immunohistochemistry staining showing clear CD68 expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.

Journal: bioRxiv

Article Title: Monocytes Strongly Induce (MYO)Fibroblast Contraction in a New 3D Skin Model to Understand the Inflammation-Fibrosis Axis in Systemic Sclerosis

doi: 10.64898/2026.02.12.705496

Figure Lengend Snippet: ( A ) Representative immunohistochemistry staining showing clear CD68 expression in the monocyte-containing hydrogels (right). Black arrows indicate a “surrounding layer” formed by the CD68 + cells covering the fibroblasts. ( B ) Flow cytometry analyses showing the percentage of total macrophages (CD45 + CD68 + ) isolated from the hydrogels after enzymatic digestion and subsequently CD45 + magnetic selection. Cells (CD45 + CD68 + ) were classified into M1-like (CD163 − ) or M2-like (CD163 + ) macrophages. Comparisons were performed using Student’s t-test (*p<0.05). Each shape represents the average of 3 technical replicates of 1 PBMC donor, n = 3.

Article Snippet: Sections were then incubated with primary rabbit –anti-human alpha-smooth muscle actin (α-SMA) (1:200) (ab5694, Abcam, UK), – fibroblast activation protein (FAP) (1:100) (ab207178, Abcam), – Phospho-SMAD2 (1:100) (#3108L, CellSignaling, USA), – Phospho-Stat3 (1:50) (#9145, CellSignaling), or mouse anti-human – CD68 (#MCA1815, BioRad, USA) (1:200) antibodies, for 1 hour at room temperature.

Techniques: Immunohistochemistry, Staining, Expressing, Flow Cytometry, Isolation, Selection