Journal: European journal of medical research

Article Title: Olfactory mucosa-mesenchymal stem cells with overexpressed Nrf2 modulate angiogenesis and exert anti-inflammation effect in an in vitro traumatic brain injury model.

doi: 10.1186/s40001-025-02344-6

Figure Lengend Snippet: Fig. 4 Effects of OM-MSCsNrf2 on the microglial polarization in BV2 cells. A–D CD86 MFI (A, B) and CD206 MFI (C, D) in BV2 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCsNrf2. scale bar: 100 μm. E–H Relative mRNA levels of Cd80 (E), Nos2 (F), Arg1 (G), and Fizz1 (H) in BV2 cells following the co-culture with OM-MSCs (ctrl) and OM-MSCsNrf2. Results of independent triplicates were expressed as mean ± standard deviation and the statistical difference between two groups was marked with asterisks (*p < 0.05, **p < 0.01, and ***p < 0.001). OM-MSCs olfactory mucosa-mesenchymal stem cells, Nrf2 Nuclear Factor Erythroid-Derived 2-Like 2, MFI mean fluorescence intensity, Nos2 nitric oxide synthase 2, Arg1: arginase 1

Article Snippet: BV2 cells after the co-culture were fixed in 4% paraformaldehyde (#P1110, Solarbio® Life Sciences, China), treated with 100% pre-chilled methanol (#34885, Sigma, Germany) and incubated with 1% bovine serum albumin (#A8010, Solarbio® Life Sciences, China) for 1 h. Next, the cells were incubated with the following primary antibodies against TMEM119 (label: Alexa Fluor® 647, #ab225494, 1:500, Abcam, UK), CD86 (label: FITC, #NBP2-34569F, 1:500, Novus Biologicals, USA), and CD206 (label: Alexa Fluor® 488, #141709, 1:500, Bio-Legend, Inc., USA) at 4°C overnight.

Techniques: Co-Culture Assay, Standard Deviation, Derivative Assay, Fluorescence

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