Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 1 | M2 macrophages (CD163+) infiltration increased with liver fibrosis progression. (A) The degree of liver fibrosis in different patients was assessed by hematoxylin-eosin staining according to the Metavir score system. F1, fibrosis around the portal vein; F2, fibrous interval around the portal vein; F3, a large number of (Continued)

Article Snippet: The slices were probed with primary antibody targeted against human CD68, CD163 (Zsbio, China), and Frontiers in Medicine | www.frontiersin.org 2 February 2021 | Volume 8 | Article 627927 CCL2 (Sigma Aldrich, St. Louis, MO, USA), and stained with either diaminobenzidine or 3-amino-9-ethylcarbazole using the Envision System (Dako Cytomation, Glostrup, Denmark).

Techniques: Staining

Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 2 | Either the co-culture condition with aHSCs or their supernatant could independently induce the M2 macrophage differentiation. After 5 days culture, compared to the control group (M0 NC), the co-aHSC group highly expressed M2 phenotype specific proteins: CD163 (29.5 ± 6.1% vs. 2.7 ± 1.1%, P < 0.001) and CD206 (28.0 ± 4.2% vs. 2.4 ± 1.2%, P < 0.001). The aHSC supernatant group independently up-regulated the expression of CD163 and CD206 on macrophages as compared to the M0 NC group (26.1 ± 2.8% vs. 2.7 ± 1.1%, P < 0.001 and 25.8 ± 3.8% vs. 2.4 ± 1.2%, P < 0.001). These experiments were repeated at least three times.

Article Snippet: The slices were probed with primary antibody targeted against human CD68, CD163 (Zsbio, China), and Frontiers in Medicine | www.frontiersin.org 2 February 2021 | Volume 8 | Article 627927 CCL2 (Sigma Aldrich, St. Louis, MO, USA), and stained with either diaminobenzidine or 3-amino-9-ethylcarbazole using the Envision System (Dako Cytomation, Glostrup, Denmark).

Techniques: Co-Culture Assay, Control, Expressing

Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 3 | The high level of CCL2 in aHSCs was associated with CD163+ macrophage infiltration and increased with liver fibrosis progression. (A) The primary aHSCs are typically fusiform and express the activation marker α-SMA, together with a high expression of CCL2 protein. (B) The aHSCs secrete high levels of CCL2 (Continued)

Article Snippet: The slices were probed with primary antibody targeted against human CD68, CD163 (Zsbio, China), and Frontiers in Medicine | www.frontiersin.org 2 February 2021 | Volume 8 | Article 627927 CCL2 (Sigma Aldrich, St. Louis, MO, USA), and stained with either diaminobenzidine or 3-amino-9-ethylcarbazole using the Envision System (Dako Cytomation, Glostrup, Denmark).

Techniques: Activation Assay, Marker, Expressing

Journal: Frontiers in medicine

Article Title: Activated Hepatic Stellate Cells Induce Infiltration and Formation of CD163 + Macrophages via CCL2/CCR2 Pathway.

doi: 10.3389/fmed.2021.627927

Figure Lengend Snippet: FIGURE 5 | TGF-β-stimulated LX2 up-regulated the expression of CD163 and CD206 on macrophages under co-culture or supernatant treatment condition. (A) The representative flow cytometry figure of macrophage phenotypic change when exposed to co-culture with LX2 or TGF-β stimulated LX2 (aLX2) or their supernatant (Continued)

Article Snippet: The slices were probed with primary antibody targeted against human CD68, CD163 (Zsbio, China), and Frontiers in Medicine | www.frontiersin.org 2 February 2021 | Volume 8 | Article 627927 CCL2 (Sigma Aldrich, St. Louis, MO, USA), and stained with either diaminobenzidine or 3-amino-9-ethylcarbazole using the Envision System (Dako Cytomation, Glostrup, Denmark).

Techniques: Expressing, Co-Culture Assay, Cytometry

Journal: Stem Cell Research & Therapy

Article Title: Human uncultured adipose-derived stromal vascular fraction shows therapeutic potential against osteoarthritis in immunodeficient rats via direct effects of transplanted M2 macrophages

doi: 10.1186/s13287-024-03946-3

Figure Lengend Snippet: A Immunohistochemical analysis of F4/80 in synovial tissue showing representative samples at 4 and 8 weeks after DMM. The sections were counterstained with hematoxylin. Scale bar = 100 μm. B F4/80 scores in each group (n = 5/group). C Immunohistochemical analysis of CD86 in synovial tissue showing representative samples at 4 and 8 weeks after DMM. The sections were counterstained with hematoxylin. Scale bar = 100 μm. D Immunohistochemical analysis of CD163 in synovial tissue showing representative samples at 4 and 8 weeks after DMM. The sections were counterstained with hematoxylin. Scale bar = 100 μm. ( E ) CD86/CD163 expression ratio (M1/M2 ratio) in each group (n = 5/group). SVF, stromal vascular fraction; ADSC, adipose-derived stromal cell; DMM, destabilization of the medial meniscus; MΦ, macrophages

Article Snippet: The samples were incubated overnight at 4 °C with a conjugated primary antibody against F4/80 (1:100; orb466169-FITC, Biorbyt), conjugated primary antibody against CD163 (1:200; BS-2527R-A594, Bioss), and primary anti-mouse antibody against hNA (1:500; ab191181, Abcam).

Techniques: Immunohistochemical staining, Expressing, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Human uncultured adipose-derived stromal vascular fraction shows therapeutic potential against osteoarthritis in immunodeficient rats via direct effects of transplanted M2 macrophages

doi: 10.1186/s13287-024-03946-3

Figure Lengend Snippet: A Immunofluorescence analysis of F4/80, CD163, and hNA in synovial tissue of the rats at 4 weeks after DMM showing representative triple immunostaining of F4/80 (green), CD163 (red), and hNA (violet) in each group. Scale bar = 20 μm. B Comparison of the ratio of CD163- and hNA-positive cells in the F4/80-positive cells via immunofluorescence staining (n = 5/group). SVF, stromal vascular fraction; ADSC, adipose-derived stromal cell; hNA, human nuclear antigen

Article Snippet: The samples were incubated overnight at 4 °C with a conjugated primary antibody against F4/80 (1:100; orb466169-FITC, Biorbyt), conjugated primary antibody against CD163 (1:200; BS-2527R-A594, Bioss), and primary anti-mouse antibody against hNA (1:500; ab191181, Abcam).

Techniques: Immunofluorescence, Triple Immunostaining, Comparison, Staining, Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Human uncultured adipose-derived stromal vascular fraction shows therapeutic potential against osteoarthritis in immunodeficient rats via direct effects of transplanted M2 macrophages

doi: 10.1186/s13287-024-03946-3

Figure Lengend Snippet: A Flow cytometry analysis revealing the proportion of CD163-positive cells in the SVF (n = 5). B Setup of the separated pellet co-culture system. Groups including ADSC, M2Φ, and SVF were established, with each administered cell type in membrane plates and OA chondrocytes in 15 mL tubes. A control group was established with no cells in membrane plates and only OA chondrocytes in 15-mL tubes. C Gross photographs and safranin-O staining of the resulting pellets in each group. D Comparison of pellet size among each group (n = 5/group). E Analysis of TGF-β, IL-10, and MMP-13 levels in the supernatant following coculture with ADSC, M2Φ, and SVF and the chondrocyte (n = 5/group). SVF, stromal vascular fraction; M2Φ, M2 macrophages; ADSC, adipose-derived stromal cell; TGF-β, transforming growth factor-β; IL-10, interleukin-10; MMP-13, matrix metalloproteinase 13

Article Snippet: The samples were incubated overnight at 4 °C with a conjugated primary antibody against F4/80 (1:100; orb466169-FITC, Biorbyt), conjugated primary antibody against CD163 (1:200; BS-2527R-A594, Bioss), and primary anti-mouse antibody against hNA (1:500; ab191181, Abcam).

Techniques: Flow Cytometry, Co-Culture Assay, Membrane, Control, Staining, Comparison, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Schematic of the proposed mechanism by which interleukin (IL)-18 amplifies macrophage (Mφ) M2 polarization and its morphological alteration, leading to excessive angiogenesis. IL-18 amplifies IL-10-induced increases in the production of osteopontin (OPN) and thrombin as soluble mediators derived from Mφ, yielding the generation of thrombin-cleaved form of OPN (Thr-OPN). Subsequently, Thr-OPN binds to integrins α4/α9 receptors on Mφ, which in turn augments M2 polarization of Mφ with higher expression of CD163 and its morphological alteration. Furthermore, CD163 may be responsible for mediating the direct cell–cell interaction between these Mφs and endothelial cells, ultimately resulting in the excessive angiogenesis.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Derivative Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Interleukin (IL)-18 amplifies macrophage (Mφ) M2 polarization and angiogenic capacity. (A) Representative FACS density plots for the expression of CD86 and CD163 in each Mφ subset. Upper, Mφ (–); middle; Mφ [tumor necrosis factor (TNF)-α]; lower, Mφ (IL-10). The numbers in each quartile of the plots are percentages of each cell population. (B) Relative mean fluorescence intensities (MFIs) of CD54, CD86, CD163, and CD206 in each Mφ subset were measured by FACS analysis, n = 3 (*** p < 0.001, * p < 0.05 vs. untreated, ## p < 0.01 vs. IL-10 alone). (C) Relative MFI of IL-18Rβ in each Mφ subset was determined by FACS analysis, n = 4 (*** p < 0.001 vs. untreated, ### p < 0.001 vs. IL-10 alone). (D,E) The total areas and lengths of tube-like structures measured by the Matrigel tube formation assay where b.End5 was cocultured with each Mφ subset, n = 6 (*** p < 0.001, ** p < 0.01, * p < 0.05 vs. untreated, ## p < 0.01, # p < 0.05 vs. IL-10 alone). (F) Representative pictures of tube-like structures visualized by calcein acetoxymethylester staining. Scale bar represents 100 µm. All data are presented as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Expressing, Fluorescence, Tube Formation Assay, Staining

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Osteopontin (OPN) drives enhancement in macrophage (Mφ) M2 polarization and angiogenic capacity. (A) Representative images of protein expression profiles obtained by comprehensive protein array in each Mφ subset. Red arrowheads indicate OPN. (B) The mRNA expression level of Spp1 relative to glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) was analyzed by real-time reverse transcription polymerase chain reaction in each Mφ subset and was normalized to Mφ (–), n = 6 [*** p < 0.001 vs. untreated, # p < 0.05 vs. interleukin (IL)-10 alone]. (C) The protein expression level of OPN relative to GAPDH was measured by western blotting and was normalized to Mφ (–), n = 10. Lower panels are typical images of each protein (*** p < 0.001 vs. untreated, # p < 0.05 vs. IL-10 alone). (D) Representative confocal laser scanning immunofluorescence overlay images of OPN (red) and DAPI (blue) in each Mφ subset. Scale bar represents 20 µm. Images in the right row are magnified regions from white or yellow rectangles in the panels of corresponding groups. Scale bar represents 10 µm. (E) Relative mean fluorescence intensity (MFI) of CD163 was measured by FACS analysis in each Mφ subset. An anti-OPN antibody (Ab) and its isotype-matched control Ab were used at 3 µg/mL, n = 4 (*** p < 0.001 vs. untreated, ## p < 0.01, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). (F) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 was cocultured with each Mφ subset, n = 12 (*** p < 0.001, ** p < 0.01, * p < 0.05 vs. untreated, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Expressing, Protein Array, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunofluorescence, Fluorescence, Tube Formation Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Thrombin contributes to macrophage (Mφ) M2 polarization and angiogenic capacity through proteolytic modification for osteopontin (OPN). (A) The mRNA expression level of Prothrombin relative to glyceraldehyde-3-phosphate dehydrogenase ( Gapdh ) was analyzed by reverse transcription polymerase chain reaction in each Mφ subset and were normalized to Mφ (–), n = 7 [*** p < 0.001, ** p < 0.01 vs. untreated, ## p < 0.01 vs. interleukin (IL)-10 alone]. (B,C) The protein expression levels of (B) thrombin or (C) OPN N-Half relative to GAPDH were measured by western blotting in each Mφ subset and were normalized to Mφ (–). Lower panels are typical images of each protein. (B) n = 8 (*** p < 0.001, * p < 0.05 vs. untreated, # p < 0.05 vs. IL-10 alone). (C) n = 16 (*** p < 0.001, * p < 0.05 vs. untreated, ## p < 0.01, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). (D) Representative confocal laser scanning immunofluorescence images of OPN (red), thrombin (green), and their merge with DAPI (blue) in each Mφ subset. Scale bar represents 20 µm. Higher magnification images are from the white rectangle region in merged panel of Mφ (IL-10 + IL-18). Scale bar represents 10 µm. (E) Relative mean fluorescence intensity (MFI) of CD163 was measured by FACS analysis in each Mφ subset. Hirudin, a specific thrombin inhibitor, was used at 1 µg/mL, n = 3 (*** p < 0.001 vs. untreated, ### p < 0.001 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). (F) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 were cocultured with each Mφ subset. Hirudin was used at 1 µg/mL, n = 6 (*** p < 0.001, ** p < 0.01 vs. untreated, ## p < 0.01, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Modification, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunofluorescence, Fluorescence, Tube Formation Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: Intergins α4/α9 are responsible for the action of osteopontin (OPN) in the augmentation of macrophage (Mφ) M2 polarization and angiogenic capacity. (A) Relative mean fluorescence intensities (MFIs) of integrins α4/α9 were measured by FACS analysis in each Mφ subset. Anti-integrin α4 or α9 antibodies (Abs) and each isotype-matched control Ab were used at 10 µg/mL, n = 4 [*** p < 0.001 vs. untreated, ### p < 0.001 vs. interleukin (IL)-10 alone]. (B) Representative confocal laser scanning immunofluorescence overlay images of integrin α4 (green) and DAPI (blue), as well as those of integrin α9 (red) and DAPI (blue) in Mφ (–) and Mφ (IL-10 + IL-18). Scale bar represents 20 µm. (C) Relative MFI of CD163 was measured by FACS analysis in each Mφ subset. Anti-integrin α4 or α9 Abs and each isotype-matched control Ab were used at 10 µg/mL, n = 4 (*** p < 0.001 vs. untreated, ### p < 0.001 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). (D) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 was cocultured with each Mφ subset. Anti-integrin α4 or α9 Abs and each isotype-matched control Ab were used at 10 µg/mL, n = 6 (*** p < 0.001, ** p < 0.01, * p < 0.05 vs. untreated, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). Integrin α4 = ITGA4; Integrin α9 = ITGA9. All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Fluorescence, Immunofluorescence, Tube Formation Assay

Journal: Frontiers in Immunology

Article Title: Interleukin-18 Amplifies Macrophage Polarization and Morphological Alteration, Leading to Excessive Angiogenesis

doi: 10.3389/fimmu.2018.00334

Figure Lengend Snippet: CD163 is a critical factor for determining the angiogenic capacity of macrophage (Mφ). (A) Upper; representative confocal laser scanning immunofluorescence overlay images of CD163 (green) and DAPI (blue) in Mφ (–) and Mφ [interleukin (IL)-10 + IL-18]. Scale bar represents 20 µm. Lower; Three-dimensional images of each upper panel. Higher magnification image in the panel of Mφ (IL-10 + IL-18) is from white rectangle region. Scale bar represents 10 µm. White arrowheads indicate CD163 highly expressed and localized at pseudopodia. (B) The total areas and lengths of tube-like structures were determined by the Matrigel tube formation assay where b.End5 were cocultured with each Mφ subset. An anti-CD163 antibody (Ab) and its isotype-matched control Ab were used at 4 µg/mL, n = 3 (*** p < 0.001, * p < 0.05 vs. untreated, ### p < 0.001, ## p < 0.01, # p < 0.05 vs. IL-10 alone, ††† p < 0.001 vs. IL-10 + IL-18). (C) Representative images (upper) and corresponding three-dimensional images (lower) of tube-like structures as well as (D) total areas and lengths of tubular structure where b.End5 (green) and Mφs (IL-10 + IL-18) (red) were cocultured on Matrigel for 16 h with an anti-CD163 Ab or its isotype-matched control Ab. Scale bar represents 100 µm, n = 8 (*** p < 0.001 vs. untreated, ### p < 0.001 vs. IL-10 + IL-18). All data are expressed as means ± SEM and were analyzed by a one-way ANOVA followed by Tukey’s test.

Article Snippet: Cells were then stained with anti-mouse Abs against phycoerythrin (PE)-conjugated CD 54 (60 ng; BioLegend, 116108), allophycocyanin (APC)-conjugated CD86 (6.0 ng; Miltenyi Biotec, 130-102-558), fluorescein isothiocyanate (FITC)-conjugated CD68 (25 ng; Bio-Rad Laboratories, MCA1957F), FITC-conjugated CD163 (200 ng; Bioss, bs-2527R-FITC), FITC-conjugated CD206 (200 ng; Bio-Rad Laboratories, MCA2235F), APC-conjugated M-CSF1 receptor (M-CSF1R) (60 ng; BioLegend, 135510), Alexa Fluor 647-conjugated IL-18Rβ (80 ng; Bioss, bs-2616R-A647), FITC-conjugated integrin α4 (130 ng; Thermo Fisher Scientific, 11-0492-82), or PE-conjugated integrin α9 (0.50 µL; Thermo Fisher Scientific, PA5-46896) for 30 min at 4°C.

Techniques: Immunofluorescence, Tube Formation Assay

Journal: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: The surface maker for the antibody used in this study.

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: Expressing, Marker

Journal: Materials Today Bio

Article Title: Vascular tissue reconstruction by monocyte subpopulations on small-diameter acellular grafts via integrin activation

doi: 10.1016/j.mtbio.2023.100847

Figure Lengend Snippet: Surface marker analysis of captured cells and porcine monocyte. The captured cells were isolated from the 3 h transplantation P-graft. a , Expression levels of CD31, CD34, CD105 and Flk-1 in CC-3H were indicated. b , Two-dimensional expression patterns of CD163/CD14 and CD16/CD14 in the captured cells were plotted. c , Two-dimensional expression patterns of CD163/CD14 and CD16/CD14 in the porcine monocyte were plotted. The MoN and MoP populations were expressed CD14 Low /CD16 Low /CD163 + and CD14 + /CD16 + /CD163 + , respectively.

Article Snippet: The cells were stained with PE-conjugated anti-CD31 antibody (MCA1746PET, Bio-Rad Laboratories, Montreal, Quebec), anti-CD34 antibody (bs-0646R-PE, Bioss Antibody Inc.), anti-CD105 antibody (bs-0579R-PE, Bioss Antibody Inc.), and anti-Flk-1 antibody (bs-0565R-PE, Bioss Antibody Inc), anti-CD163 antibody (bs-2527R-PE, Bioss Antibody Inc.), anti-CD14 antibody (MCA1218F, Bio-Rad Laboratories, Inc., Hercules, CA), and anti-CD16 antibody (MCA1971PE, Bio-Rad Laboratories, Inc.).

Techniques: Marker, Isolation, Transplantation Assay, Expressing

Journal: Brain, behavior, and immunity

Article Title: Intermittent cytomegalovirus infection alters neurobiological metabolism and induces cognitive deficits in mice

doi: 10.1016/j.bbi.2023.12.033

Figure Lengend Snippet: Brain cell characterization flow panel.

Article Snippet: CD163 , AF680 , Bioss , bs-2527R-A680 , 1:400.

Techniques:

Journal: The Journal of steroid biochemistry and molecular biology

Article Title: Deletion of JNK2 prevents vitamin-D-deficiency-induced hypertension and atherosclerosis in mice

doi: 10.1016/j.jsbmb.2017.09.014

Figure Lengend Snippet: JNK2−/− LDLR−/− and LDLR−/− mice were fed vitamin D-sufficient or deficient high fat diet for 10 weeks (n=4M/4F mice per group). Peritoneal macrophages harvested after HFD were assessed for (A) flow cytometry of membrane expression of M1 macrophage markers (CCR7 and CD86) and M2 markers (MR and CD163) and (B–D) mRNA expression of macrophage cytokines. Data are expressed as mean ± SEM, and horizontal lines demonstrate comparison between individual groups where *p≤0.05, **p≤0.01, ***p≤0.001.

Article Snippet: Macrophage cell surface marker analysis was performed using a FACStar Plus with PE-conjugated anti-CCR7 and anti-CD86 (E-Bioscience) for M1 macrophage membrane protein expression and FITC-conjugated anti-CD163 (Bioss USA) and anti-MR (R&D Systems) for M2 macrophage membrane protein expression in unstimulated peritoneal macrophages collected from HFD-fed mice [ 28 , 39 ].

Techniques: Flow Cytometry, Expressing