primary antibodies against cd86  (Bioss)

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: Effects of rTMS on neuroinflammation and the M1/M2 polarization state of microglia in vivo . (A–C) Elisa was used to examine the expression levels of IL-6, TNF-α and IL-10 expression level. (D–F) Real-time qRT-PCR analysis of the relative mRNA expression of TMEM119 , iNOS , and Arg1 in the DLPFC. (G, H) Immunofluorescence staining of Iba-1, M1-type (CD86) and M2-type (CD206) markers in the left PFC region. (I, J) Proportion of positive cells for target proteins. The number of samples in each group is 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: Primary antibodies against CD86 (1:100; bs-1035r; Bioss), CD206 (1:100; #24595; CST) and Iba-1(1:100, ab283319, Abcam, Cambridge, UK)were applied, and the sections were incubated overnight at 4 °C.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: Effects of rTMS on neuroinflammation and the M1/M2 polarization state of microglia in vitro . (A–D) Elisa was used to examine the expression levels of NLRP3, IL-6, TNF-α and IL-10 expression level. (E–G) Immunofluorescence staining of Iba-1, CD86, CD206. (H–J) Proportion of positive cells for target proteins. The number of samples in each group was 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01 and ****P < 0.0001.

Article Snippet: Primary antibodies against CD86 (1:100; bs-1035r; Bioss), CD206 (1:100; #24595; CST) and Iba-1(1:100, ab283319, Abcam, Cambridge, UK)were applied, and the sections were incubated overnight at 4 °C.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: The roles of METTL3 and NMDAR2B in the regulation of microglial cell polarization by magnetic stimulation in vitro . (A–C, G–I) Immunofluorescence staining of Iba-1, CD86 and CD206. (D–F, J–L) Proportion of positive cells for target proteins. The number of samples in each group was 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: Primary antibodies against CD86 (1:100; bs-1035r; Bioss), CD206 (1:100; #24595; CST) and Iba-1(1:100, ab283319, Abcam, Cambridge, UK)were applied, and the sections were incubated overnight at 4 °C.

Techniques: In Vitro, Immunofluorescence, Staining

Journal: Journal of Clinical and Translational Hepatology

Article Title: Arctigenin Prevents Metabolic Dysfunction-associated Steatohepatitis by Inhibiting NLRP3/GSDMD-N Axis in Macrophages

doi: 10.14218/JCTH.2025.00141

Figure Lengend Snippet: (A) Chemical structure of ATG. (B) Target prediction of ATG. (C) Molecular targets associated with MAFLD. (D) Intersection of ATG-specific and MAFLD-related targets. (E) The PPI network of intersection targets between ATG and MAFLD-related targets. (F) GO enrichment analysis of the intersected targets. (G–H) Enrichment of KEGG pathway (G) and Reactome pathway (H) of ATG’s targets. (I–J) Immunofluorescence analysis of the colocalization of NLRP3 and CD86 in mice liver from four groups (scale bar: 20 µm), staining intensity was quantified, and colocation analysis was performed. (K) NLRP3 with ATG bound to the cavity. (L) Western blot of CETSA experiment to further confirm the interaction between NLRP3 and ATG in RAW264.7 cells, the temperature ranges from 49°C to 61°C, with relative band intensity normalized to 49°C. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ATG, arctigenin; MAFLD, metabolic dysfunction-associated fatty liver disease; PPI, protein-protein interaction; KEGG, Kyoto Encyclopedia of Genes and Genomes; NLRP3, NLR family pyrin domain containing 3; SEM, standard error of the mean.

Article Snippet: Primary antibodies against CD86 (19589, Cell Signaling Technology, CST®; 1:400 in 3% BSA) and α-SMA (ab124964, Abcam®; 1:2,000 in 3% BSA) were incubated overnight.

Techniques: Immunofluorescence, Staining, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: Construction of cartilaginous organoids based on cartilage extracellular matrix microcarriers to promote articular cartilage regeneration through immune regulation

doi: 10.1016/j.jot.2025.05.005

Figure Lengend Snippet: Effect of CEMMS on the polarization of rat macrophages. A) Quantitative analysis of M1 macrophage-related phenotypes using real-time PCR (n = 3). B) Quantitative analysis of M2 macrophage-related phenotypes using real-time PCR (n = 3). C) Immunofluorescence staining for markers of macrophages(CD86; Green)/(CD206; Red). Scale bars: 100 μm. D) Quantitative analysis of immunofuorescence staining. E) Immunofluorescence staining for markers of macrophages (Arg-1; Green)/(IL-1β; Red). Scale bars: 100 μm. F) Quantitative analysis of immunofuorescence staining. ∗∗∗∗ p < 0.0001.

Article Snippet: The cells were incubated with primary antibodies against CD86 (NBP2-25208, Novus), CD206 (24595S, CST), IL-1β(ab254360, Abcam) and Arg-1 (66129-1-Ig, proteintech) overnight at 4 °C.

Techniques: Real-time Polymerase Chain Reaction, Immunofluorescence, Staining