Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence and western blotting results showing the expression of relevant cytokines in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing ANGPT2 + , Tie2 + , and CD11b + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of ANGPT2 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Counts of TEMs (Tie2 + and CD11b + ) in each group (n = 9, one-way ANOVA and Šídák's multiple comparisons test). D Representative western blot showing the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 in the CIBT adjacent to the TM tissue in each group (normalized to β-actin expression). E Densitometric analyses of the relative expression of TRPS1, ANGPT2, Tie2, CD11b, VEGFA, IGF1, and CD31 (n = 3, one-way ANOVA and Šídák's multiple comparisons test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001). The error bars represent the ± SDs. CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Western Blot, Expressing, Staining

Journal: Cell & Bioscience

Article Title: Tie2-expressing monocytes/macrophages promote angiogenesis in chronically ischaemic brain tissue

doi: 10.1186/s13578-025-01401-1

Figure Lengend Snippet: Immunofluorescence results showing the expression of pro-angiogenesis factors in the CIBT of 2VO + EMS rats. A Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and VEGFA + cells and in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. B Counts of VEGF + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). C Representative triple immunofluorescence staining images showing Tie2 + , CD11b + , and IGF1 + cells in the CIBT in each group (white arrows indicate TEMs). Bar = 50 µm. D Counts of IGF1 + cells in each group (n = 3, one-way ANOVA and Šídák's multiple comparisons test). CIBT: chronically ischaemic brain tissue; EMS: encephalomyosynangiosis; TEMs: Tie2-expressing monocytes/macrophages; 2VO: 2-vessel occlusion

Article Snippet: After being washed with PBST, the slices were incubated with 488-conjugated goat anti-rabbit (1:2000, ab150077, Abcam, UK), 594-conjugated goat anti-mouse (1:2000, ab150116, Abcam, UK), and 647-conjugated donkey anti-goat (1:2000, ab150135, Abcam, UK) secondary antibodies at room temperature in a dark room for 2 h. The 647-conjugated CD11b antibody (0.5 μg, CL647-65229, proteintech, China) was used to directly label CD11b.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Frontiers in Immunology

Article Title: Unfolded Protein Response Differentially Regulates TLR4-Induced Cytokine Expression in Distinct Macrophage Populations

doi: 10.3389/fimmu.2019.01390

Figure Lengend Snippet: Resident macrophages from liver are sensitive to stress-induced change in cytokine mRNA expression. (A,B) WT (A) and TRIF-/- (B) mice were challenged with DMSO or Tm (1.25 mg/kg body weight) for 18 h prior to the isolation of non-parenchymal cells from the liver. The non-parenchymal cells were immunostained for surface CD11b, Ly6C, and F4/80 and subjected to flow cytometry analysis. (C) The percentage of LY6C+ and F4/80+ cells within the gated populations isolated from livers of mice treated with DMSO vs. Tm ( n = 4). (D–F) Liver non-parenchymal cells from WT or TRIF-/- mice treated with DMSO or Tm were labeled with anti-CD11b-PE antibody and then isolated using anti-PE magnetic microbeads. CD11b+ cells were subsequently treated with or without LPS (100 ng/ml) for 3 h. Total RNA was used to determine IL12p35 (D) , IL12p40 (E) , and IL10 (F) mRNAs by real time PCR. Data are presented as the mean ± SD of triplicate experiments for WT ( n = 3 per treatment for each experiments) and duplicate experiments for TRIF-/- mice ( n = 3 per treatment for each experiments). The differences between indicated treated samples were evaluated by two way-ANOVA and Turkey's multiple comparisons test. * p < 0.05.

Article Snippet: Anti-Ly6C-AlexaFluor (clone ER-MP20) was obtained from Biorad.

Techniques: Expressing, Isolation, Flow Cytometry, Labeling, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: Unfolded Protein Response Differentially Regulates TLR4-Induced Cytokine Expression in Distinct Macrophage Populations

doi: 10.3389/fimmu.2019.01390

Figure Lengend Snippet: Sensitivity to stress-induced modulation of cytokine expression in infiltrating liver myeloid cells varies over the course of inflammatory response. (A) WT Mice were injected i.p. with APAP (300 mg/kg body weight) for 24 or 72 h prior to the isolation of liver non-parenchymal cells. The isolated liver non-parenchymal cells were immunostained for surface CD11b, CD206, and Ly6C and analyzed by flow cytometry. (B) The percentage of LY6C+ and CD206+ cells within the gated cell populations ( n = 4). (C–E) WT and TRIF-/- mice were injected i.p. with APAP (300 mg/kg) for 24 or 72 h and treated with DMSO or Tm i.p. during the final 18 h. Liver CD11b+ cells from WT and TRIF-/- mice were isolated using anti-CD11b-PE and MACS magnetic column followed by treatment with or without LPS (100 ng/ml) for 3 h. Total RNA was prepared and used to determine IL12p35 (C) , IL12p40 (D) , and IL10 (E) mRNA levels by real time PCR. Data are presented as the mean ± SD of triplicate experiments for WT ( n = 3 per treatment for each experiments) and duplicate experiments for TRIF-/- mice ( n = 3 per treatment for each experiments). The differences between indicated treatments were evaluated by two way-ANOVA and Turkey's multiple comparisons test. * p < 0.05 was considered significant.

Article Snippet: Anti-Ly6C-AlexaFluor (clone ER-MP20) was obtained from Biorad.

Techniques: Expressing, Injection, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction