anti 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: Frontiers in Immunology

Article Title: TENS alleviates CP/CPPS-related inflammation and pain by modulating Kir2.1-dependent macrophage polarization

doi: 10.3389/fimmu.2025.1683500

Figure Lengend Snippet: The ratio of CD86 + M1 macrophages is positively correlated with prostate tissue inflammation in patients. (a) Representative H&E images depicting the evaluation of inflammation grades in prostate tissue. (b , c) Representative IHC images of CD86 + M1 macrophages and CD206 + M2 macrophages in prostate tissue of patients with mild, moderate, and severe inflammation, respectively. Scale bars: 50 μm. (d) Quantitative analyses of CD86 + M1 macrophages, CD206 + M2 macrophages, and the M1/M2 ratio in prostate tissue. CD86 + M1 and CD206 + M2 cells were counted per high-power field (HPF) at × 200 magnification. (e) Spearman’s rank correlation analysis between inflammation scores and CD86 + cell counts, CD206 + cell counts, or M1/M2 ratios. Data are shown as the mean ± SEM ( n = 9 mild, 11 moderate, 6 severe) and were analyzed using a two-tailed unpaired Student’s t -test. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs . the mild group.

Article Snippet: Sections were incubated overnight at 4°C with primary antibodies against CD86 (bs-1035R, Bioss, Beijing, China), CD206 (18704-1-AP, Proteintech, Wuhan, China), TNF-α ( GB115701 , Servicebio, Wuhan, China), IL-1β (GB11113, Servicebio, Wuhan, China), IL-6 (GB11117, Servicebio, Wuhan, China), Cyclooxygenase-2 (COX-2) (Abcam, ab179800, Cambridge, UK), substance P (SP; Proteintech, Wuhan, China, 13839-1-AP), or Kir2.1 (19965-1-AP, Proteintech, Wuhan, China).

Techniques: Two Tailed Test

Journal: Frontiers in Immunology

Article Title: TENS alleviates CP/CPPS-related inflammation and pain by modulating Kir2.1-dependent macrophage polarization

doi: 10.3389/fimmu.2025.1683500

Figure Lengend Snippet: TENS induced M2 polarization of macrophages in prostate tissue. (a) Representative IHC images of CD86 + M1 macrophages and CD206 + M2 macrophages in rat prostate tissue. Scale bars: 50 μm. (b) Quantitative analyses of CD86 + M1 macrophages, CD206 + M2 macrophages, and the M1/M2 ratio in prostate tissue. CD86 + M1 and CD206 + M2 cells were counted per high-power field (HPF) at × 200 magnification. Data are shown as the mean ± SEM and were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. # p < 0.05 and ## p < 0.01 vs . the control group. * p < 0.05 and ** p < 0.01 vs . the EAP group ( n = 4 per group).

Article Snippet: Sections were incubated overnight at 4°C with primary antibodies against CD86 (bs-1035R, Bioss, Beijing, China), CD206 (18704-1-AP, Proteintech, Wuhan, China), TNF-α ( GB115701 , Servicebio, Wuhan, China), IL-1β (GB11113, Servicebio, Wuhan, China), IL-6 (GB11117, Servicebio, Wuhan, China), Cyclooxygenase-2 (COX-2) (Abcam, ab179800, Cambridge, UK), substance P (SP; Proteintech, Wuhan, China, 13839-1-AP), or Kir2.1 (19965-1-AP, Proteintech, Wuhan, China).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: TENS alleviates CP/CPPS-related inflammation and pain by modulating Kir2.1-dependent macrophage polarization

doi: 10.3389/fimmu.2025.1683500

Figure Lengend Snippet: Electrical stimulation promoted the repolarization of M1 macrophages into M2 macrophages. (a) Cell viability after ES was assessed using the CCK8 assay. (b) Flow cytometry analysis of CD86 + M1 and CD206 + M2 macrophages in control, LPS-treated, and ES-treated (0.1 V/cm, 0.25 V/cm) RAW264.7 cells. MFI, mean fluorescence intensity. (c) Reactive oxygen species (ROS) detection in control, LPS-treated, and ES-treated (0.1 V/cm, 0.25 V/cm) RAW264.7 cells. (d) Western blot analysis of M1 markers (CD86, iNOS, TLR4) and M2 markers (CD206, CD163, CD209) in control, LPS-stimulated, and LPS + ES-treated (0.25 V/cm) RAW264.7 cells. (e) Quantitative analyses of macrophage phenotype marker expression. (f) ELISA analyses of TNF-α, IL-1β, IL-6, and IL-10 secretion in macrophage culture supernatants. Data are shown as the mean ± SEM and were analyzed using one-way ANOVA followed by Tukey’s post-hoc test . ## p < 0.01 and ### p < 0.001 vs . the control group. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs . the LPS group ( n = 3 per group).

Article Snippet: Sections were incubated overnight at 4°C with primary antibodies against CD86 (bs-1035R, Bioss, Beijing, China), CD206 (18704-1-AP, Proteintech, Wuhan, China), TNF-α ( GB115701 , Servicebio, Wuhan, China), IL-1β (GB11113, Servicebio, Wuhan, China), IL-6 (GB11117, Servicebio, Wuhan, China), Cyclooxygenase-2 (COX-2) (Abcam, ab179800, Cambridge, UK), substance P (SP; Proteintech, Wuhan, China, 13839-1-AP), or Kir2.1 (19965-1-AP, Proteintech, Wuhan, China).

Techniques: CCK-8 Assay, Flow Cytometry, Control, Fluorescence, Western Blot, Marker, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: TENS alleviates CP/CPPS-related inflammation and pain by modulating Kir2.1-dependent macrophage polarization

doi: 10.3389/fimmu.2025.1683500

Figure Lengend Snippet: Repolarization mechanism of ES effect in macrophages. (a) GO classification of DEPs between the LPS and ES groups related to cellular component, molecular function, and biological process categories. (b) Heatmap of differentially clustered ion transmembrane transporter activity. The color scale represents the Z -score of normalized protein abundance; red indicates upregulation, and blue indicates downregulation. (c) Representative WB images of Kir2.1 and TRPV2 expression. (d) Quantitative analyses of Kir2.1 and TRPV2 marker expression. (e) Representative IHC images of Kir2.1 in rat prostate tissue. Scale bars: 50 μm. (f) Quantitative analyses of CD86 + and CD206 + expression in macrophages measured by flow cytometry. (g) Immunofluorescence staining images of cellular membrane potential in macrophages (scale bars: 20 μm). (h) Quantitative analysis of cellular membrane potential in macrophages measured by flow cytometry. (i) Immunofluorescence staining images of intracellular Ca 2+ concentration in macrophages (scale bars: 20 μm). (j) Quantitative analysis of intracellular Ca 2+ concentration in macrophages measured by flow cytometry. MFI, mean fluorescence intensity. (k) Representative WB images of NF-κB/STAT1/STAT6 signaling pathway expression. (l) Quantitative analyses of NF-κB/STAT1/STAT6 signaling pathway expression. Data are shown as the mean ± SEM and were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. ## p < 0.01 and ### p < 0.001 vs . the LPS group. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs . the ES group ( n = 3 per group).

Article Snippet: Sections were incubated overnight at 4°C with primary antibodies against CD86 (bs-1035R, Bioss, Beijing, China), CD206 (18704-1-AP, Proteintech, Wuhan, China), TNF-α ( GB115701 , Servicebio, Wuhan, China), IL-1β (GB11113, Servicebio, Wuhan, China), IL-6 (GB11117, Servicebio, Wuhan, China), Cyclooxygenase-2 (COX-2) (Abcam, ab179800, Cambridge, UK), substance P (SP; Proteintech, Wuhan, China, 13839-1-AP), or Kir2.1 (19965-1-AP, Proteintech, Wuhan, China).

Techniques: Activity Assay, Quantitative Proteomics, Expressing, Marker, Flow Cytometry, Immunofluorescence, Staining, Membrane, Concentration Assay, Fluorescence