immunofluorescence staining for cd86  (Bioss)

Journal: Bioactive Materials

Article Title: End-tail soaking strategy toward robust and biomimetic sandwich-layered hydrogels for full-thickness bone regeneration

doi: 10.1016/j.bioactmat.2025.02.045

Figure Lengend Snippet: Effects of osteoblasts on immune regulation. (a) Reactome annotation analysis of the close relation between differential genes and immune activation. (b) Analysis of PPI networks after co-culture of hydrogel and mesenchymal stem cells. (c) Expression of macrophage polarization-related genes detected by qPCR after hydrogel co-culture with macrophages (n = 3). (d) Macrophage polarization phenotype of macrophages co-cultured with hydrogel-pretreated BMSCs detected by qPCR (n = 3). (e) Detection of macrophage M2 type polarization in co-culture groups by flow cytometry. Wherein, bmsc(+) means multicellular co-culture. (f) Polarization quantification by flow cytometry of macrophages (n = 3). (g) Differential ratio of polarization of macrophages after direct versus indirect co-culture in hydrogels detected by flow cytometry. (h) Immunofluorescence staining of macrophage polarization states in animal sections (Blue: DAPI; Red:CD86; Green:CD206. Scale bar, 50 μm). The arrow indicates the interface between newly formed bone and macrophages.

Article Snippet: The immunofluorescence staining method for CD86 (GB115630, Servicebio, China), CD206 (GB113497, Servicebio, China), and CD31 (GB120005, Servicebio, China) were the same as described above.

Techniques: Activation Assay, Co-Culture Assay, Expressing, Cell Culture, Flow Cytometry, Immunofluorescence, Staining

Journal: Heliyon

Article Title: BushenHuoxue decoction suppresses M1 macrophage polarization and prevents LPS induced inflammatory bone loss by activating AMPK pathway

doi: 10.1016/j.heliyon.2023.e15583

Figure Lengend Snippet: BSHXD inhibited LPS-induced M1 polarization of RAW264.7 macrophages. (A) The morphology of RAW264.7 cells observed by light microscope. (B) Immunofluorescence staining of RAW264.7 cells. (C)The protein level of CD86, detected by Western blot analysis. (D) CD86 was normalized to β-Actin and expressed in arbitrary units. (E) The proportion of F4/80+ and CD86 + in RAW264.7 cells, detected by flow cytometry. N = 3, data are expressed as mean ± SD. *p < 0.05, **p < 0.01 compared to LPS group.

Article Snippet: We also completed immunofluorescence staining (IF) for CD86 (1:200, #19589, CST) and F4/80 (1:200, #70076, CST).

Techniques: Light Microscopy, Immunofluorescence, Staining, Western Blot, Flow Cytometry

Journal: Heliyon

Article Title: BushenHuoxue decoction suppresses M1 macrophage polarization and prevents LPS induced inflammatory bone loss by activating AMPK pathway

doi: 10.1016/j.heliyon.2023.e15583

Figure Lengend Snippet: BSHXD inhibits inflammatory factors and M1 macrophages through activating AMPK. (A) The CD86 protein level, detected by Western blot analysis. (B) CD86 was normalized to β-Actin and expressed in arbitrary units. (C) IL-1β, IL-6 and TNF-α protein levels, detected by Western blot analysis. (D) IL-1β, (E) IL-6 and (F) TNF-α were normalized to β-Actin and expressed in arbitrary units. (G) The proportion of F4/80 + and CD86 + in RAW264.7 cells, detected by flow cytometry. (H) Typical images of immunofluorescence staining: red (CD86), green (F4/80), and blue (DAPI). Scale bar indicates 100 μm.N = 3. Data are presented as mean ± SD of three independent experiments. *p < 0.05, **p < 0.01.

Article Snippet: We also completed immunofluorescence staining (IF) for CD86 (1:200, #19589, CST) and F4/80 (1:200, #70076, CST).

Techniques: Western Blot, Flow Cytometry, Immunofluorescence, Staining

Journal: Heliyon

Article Title: BushenHuoxue decoction suppresses M1 macrophage polarization and prevents LPS induced inflammatory bone loss by activating AMPK pathway

doi: 10.1016/j.heliyon.2023.e15583

Figure Lengend Snippet: BSHXD inhibited LPS-induced inflammatory bone loss. (A)Immunohistochemical staining of TNF-α in mouse skull. Scale bar indicates 200 and 50 μm. (B)Immunofluorescence staining in mouse skull: red (CD86), green (F4/80), and blue (DAPI). Scale bar indicates 100 μm. (C) Number of TNF-α positive cells. (D) Number of CD86 positive cells. N = 3. All data were expressed as the mean ± SD. *p < 0.05, **p < 0.01, compared with the vehicle group.

Article Snippet: We also completed immunofluorescence staining (IF) for CD86 (1:200, #19589, CST) and F4/80 (1:200, #70076, CST).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence