Journal: Respiratory Research

Article Title: HDAC8 inhibitor attenuates airway responses to antigen stimulus through synchronously suppressing galectin-3 expression and reducing macrophage-2 polarization

doi: 10.1186/s12931-020-1322-5

Figure Lengend Snippet: Macrophage polarization states. Distribution and proportion of CD68 + ( a ), CD86 + ( c ) and CD163 + ( e ) macrophages (brown) were identified using IHC procedures. Intensity for the color response to antibodies was visible in lung tissues from all groups. Population proportions for the stained cells as shown by red arrows were calculated as a fold change of the control. ( b , d , f ). The results were expressed as Mean ± SD ( n = 3). **: a p -value of < 0.01 vs NS, OVA/BUD and OVA/PCI except the number of CD86 + cells in OVA/PCI group. #: P < 0.05 vs NS and ##; P < 0.01 vs either NS or OVA/BUD group

Article Snippet: The sections were incubated with primary antibodies of CD68 at 1:100 (sc-20,060; Santa-Cruz Biotechnology, CA, US), CD86 at 0.5 μg/ml (NBP2–25208; Novus Biologicals, CO, US) and CD163 at 1:500 (ab182422; Abcam, Cambridge, UK) overnight at 4 °C.

Techniques: Staining, Control

Journal: Molecules (Basel, Switzerland)

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

doi: 10.3390/molecules30010123

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

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

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

Journal: Molecules (Basel, Switzerland)

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

doi: 10.3390/molecules30010123

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

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

Techniques: Expressing, Fluorescence, Marker

Journal: Molecular Biomedicine

Article Title: Deleting fibroblast growth factor 2 in macrophages aggravates septic acute lung injury by increasing M1 polarization and inflammatory cytokine secretion

doi: 10.1186/s43556-024-00203-0

Figure Lengend Snippet: Effect of FGF2 deficiency on BMDM apoptosis and polarization. a – c FGF2 deletion increased BMDM apoptosis. a Apoptosis in BMDM deprived of FBS for 24 h was assessed by flow cytometry ( n = 4). b - c Percentage of PI + Annexin V + and PI- Annexin V + BMDM after starvation. d - k FGF2 deletion in BMDM promoted M1 polarization. d - g Flow cytometric analysis of macrophage markers in BMDM treated with LPS or IL4, including CD86, iNOS, CD206, and Arg1 ( n = 3). h - k The levels of CD86, iNOS, CD206 and Arg1 in BMDM after treatment with LPS or IL4. N represents no treatment; * p < 0.05, vs. WT; Ψ p < 0.05, vs. N + WT; Ω p < 0.05, vs. N + FGF2 KO

Article Snippet: The tissue sections were incubated overnight at 4°C with primary antibodies, including the rabbit polyclonal antibody CD206 (1:200, Cat No. A02285-2, Boster, Wuhan, China), rabbit monoclonal antibody CD86 (1:100, Cat No. BM4121, Boster, Wuhan, China), rabbit polyclonal antibody F4/80 (1:100, Cat No. 29414-1-AP, Proteintech, Wuhan, China), anti-mouse NLRP3 (1:200, Cat No.68102-1-Ig, Proteintech, Wuhan, China), Caspase-1 p20 Rabbit pAb (1:400, Cat No.bs-10743R, Bioss, Beijing, China) and ASC/TMS1 Rabbit PolyAb (1:200, Cat No.10500-1-AP, Proteintech, Wuhan, China).

Techniques: Flow Cytometry

Journal: Molecular Biomedicine

Article Title: Deleting fibroblast growth factor 2 in macrophages aggravates septic acute lung injury by increasing M1 polarization and inflammatory cytokine secretion

doi: 10.1186/s43556-024-00203-0

Figure Lengend Snippet: Mice reconstituted with FGF2 KO macrophages and subjected to CLP demonstrate increased M1 polarization in lung tissue. a - f The presence and levels of CD206, CD86, and F4/80 markers on macrophages within lung tissue were identified and quantitatively assessed using immunofluorescence staining. Bar is 20 μm. * p < 0.05, vs. WT; Δ p < 0.05 vs. WT + LPS; # p < 0.05 vs. FGF2 KO

Article Snippet: The tissue sections were incubated overnight at 4°C with primary antibodies, including the rabbit polyclonal antibody CD206 (1:200, Cat No. A02285-2, Boster, Wuhan, China), rabbit monoclonal antibody CD86 (1:100, Cat No. BM4121, Boster, Wuhan, China), rabbit polyclonal antibody F4/80 (1:100, Cat No. 29414-1-AP, Proteintech, Wuhan, China), anti-mouse NLRP3 (1:200, Cat No.68102-1-Ig, Proteintech, Wuhan, China), Caspase-1 p20 Rabbit pAb (1:400, Cat No.bs-10743R, Bioss, Beijing, China) and ASC/TMS1 Rabbit PolyAb (1:200, Cat No.10500-1-AP, Proteintech, Wuhan, China).

Techniques: Immunofluorescence, Staining

Journal:

Article Title: Endotoxin-Induced Gamma Interferon Production: Contributing Cell Types and Key Regulatory Factors

doi: 10.1128/CDLI.9.3.530-543.2002

Figure Lengend Snippet: Effects of B7 inhibitors on LPS-induced IFN-γ production. Isolated splenocytes were incubated with LPS (1 μg/ml) or 108 CFU of heat-killed bacteria per ml in the presence of nonspecific goat IgG (10 μg/ml), CTLA4-Ig (1 μg/ml), anti-CD80 (10 μg/ml), or anti-CD86 (10 μg/ml) for 24 h. The IFN-γ levels in the conditioned media were determined by ELISA (n = 4 to 8 wells/group). In some experiments, anti-CD80 and anti-CD86 were preabsorbed with CD80-Ig or CD86-Ig to assess specificity. ∗, significantly (P < 0.05) less than the levels produced in the goat IgG group.

Article Snippet: CD80-Ig and CD86-Ig fusion proteins were purchased from Ancell Corporation (Bayport, Minn.).

Techniques: Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Produced

Journal:

Article Title: Endotoxin-Induced Gamma Interferon Production: Contributing Cell Types and Key Regulatory Factors

doi: 10.1128/CDLI.9.3.530-543.2002

Figure Lengend Snippet: Expression of splenic B7 proteins (CD80 and CD86) following LPS challenge. Mice were challenged with LPS (4 mg/kg of body weight, intraperitoneally), and their spleens were harvested 8 h later. Saline-injected mice served as controls. The levels of CD80 and CD86 expression on CD14+, CD19+, and CD3+ splenocytes were determined by flow cytometry.

Article Snippet: CD80-Ig and CD86-Ig fusion proteins were purchased from Ancell Corporation (Bayport, Minn.).

Techniques: Expressing, Injection, Flow Cytometry

Journal:

Article Title: Endotoxin-Induced Gamma Interferon Production: Contributing Cell Types and Key Regulatory Factors

doi: 10.1128/CDLI.9.3.530-543.2002

Figure Lengend Snippet: IFN-γ production by isolated splenic T and NK cells in response to IFN-γ-regulating factors. Isolated splenic T and NK cells were incubated with IL-12 (2 ng/ml) alone or with IL-12 plus IL-15 (20 ng/ml), IL-18 (20 ng/ml), agarose-coupled anti-CD28 (10 μg/ml), CD80-Ig (10 μg/ml), CD86-Ig (10 μg/ml), or anti-CD3 (10 μg/ml) in the indicated combinations for 24 h. The IFN-γ levels in the conditioned media were determined by ELISA (n = 4 to 8 wells/group).

Article Snippet: CD80-Ig and CD86-Ig fusion proteins were purchased from Ancell Corporation (Bayport, Minn.).

Techniques: Isolation, Incubation, Enzyme-linked Immunosorbent Assay