isolectin b4 594  (Vector Laboratories)

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: Distribution patterns of BAMs after SAH and their effects on cognitive function and myelin. A Schematic diagram of the animal groups and experimental procedures. B Representative images of immunofluorescence staining for border-associated macrophages in the meninges of mice in the sham group and at 1 day, 7 days, and 14 days after SAH (white, Lyve1; red, CD206; blue, DAPI). C Representative images of immunofluorescence staining for border-associated macrophages in the perivascular space of the cortical region in mice from the sham group and at 1 day, 7 days, and 14 days after SAH (white, lectin; red, CD206; blue, DAPI). Scale bar, 100 μm. D Quantification of CD206 + BAMs in the meninges and perivascular space of the cortical region ( n = 6 per group; one-way ANOVA). E Representative images of myelin immunofluorescence staining in the SAH-Vehicle group and the SAH-CLO Lip group with border-associated macrophage depletion by clodronate liposomes (red, MBP; green, NF200; white, PDGFRα; blue, DAPI). Scale bar, 200 μm. F , G Quantitative analysis of MBP-positive (MBP +) myelin, NF200-positive (NF200 +) axons, and NF200 + /MBP + and MBP − /NF200. + areas in panel E ( n = 6 per group; T-test). H Statistical analysis of the trend in latency from day 1 to day 5 in the Morris water maze test, as well as the time spent in the target quadrant and the number of platform crossings ( n = 6 per group, two-way ANOVA). I Statistical chart of the time spent by the mice in the new arm and the percentage of alternations in the Y-maze test ( n = 6 per group, one-way ANOVA). J Statistical chart of the proportion of open arm entries to total arm entries by the mice in the elevated plus maze test ( n = 6 per group; one-way ANOVA)

Article Snippet: After blocking with QuickBlockTM Immunostaining Blocking Solution (Shanghai Beyotime Biotechnology Co., Ltd., China, Cat. No. P0260) for 15 min, the following primary antibodies were incubated overnight at 4 °C: anti-Lyve1 antibody (5 μg/ml, mouse, Abcam, ab33682); recombinant anti-mannose receptor antibody [ EPR25215 –277] (1:50, rabbit, Abcam, ab300621); MBP antibody (F-6) (1:50, mouse, SANTA CRUZ Biotechnology, sc-271524); NF-H recombinant SuperclonalTM antibody (2 μg/ml, rabbit, Thermo Fisher, 711,025); PDGFRα antibody (5 μg/ml, goat, RD systems, AF1062); and lectin (594, vector labs, DL-1207–5).

Techniques: Immunofluorescence, Staining, Liposomes

Journal: Journal of Neuroinflammation

Article Title: Remote ischemic postconditioning improves cognitive dysfunction after subarachnoid hemorrhage by driving metabolic reprogramming of border-associated macrophages through the IL-33/ST2 axis

doi: 10.1186/s12974-025-03587-0

Figure Lengend Snippet: RIPostC treatment increases the abundance of BAMs after SAH and improves cognitive dysfunction. A Schematic diagram of the animal groups and experimental procedures. B , C Representative images of lower limb perfusion speckle in mice during the RIPostC treatment phase and statistical analysis of lower limb blood flow perfusion ROI values ( n = 6 per group; one-way ANOVA). D Representative images of meningeal border-associated macrophage immunofluorescence staining in the SAH group and SAH-RIPostC group (red, CD206; white, Lyve1; blue, DAPI). E Representative images of immunofluorescence staining of border-associated macrophages in the perivascular space of the cortical region in the SAH group and the SAH-RIPostC group (red, CD206; white, lectin; blue, DAPI). Scale bar, 100 μm. F Quantitative analysis of CD206 + BAMs in the meningeal and perivascular spaces of the cortical region, as shown in D-E ( n = 6 per group; T-test). G Representative images of myelin immunofluorescence staining in the SAH group and the SAH-RIPostC group (green, NF200; red, MBP; white, PDGFRα; blue, DAPI). Scale bar, 200 μm. H Quantitative analysis of NF200-positive (NF200 + ) axons, MBP-positive (MBP + ) myelin, and the MBP − /NF200 + and NF200 + /MBP + regions in G ( n = 6 per group; T-test). I Statistical analysis of the trend in latency from day 1 to day 5 in the Morris water maze test, as well as the time spent in the target quadrant and the number of platform crossings in both groups of mice ( n = 6 per group, two-way ANOVA and T-test). J Statistical chart of the time spent in the novel arm and the percentage of alternations in the Y-maze test ( n = 6 per group, T-test). K Statistical chart of the proportion of open arm entries to total arm entries in the elevated plus maze test ( n = 6 per group; T-test)

Article Snippet: After blocking with QuickBlockTM Immunostaining Blocking Solution (Shanghai Beyotime Biotechnology Co., Ltd., China, Cat. No. P0260) for 15 min, the following primary antibodies were incubated overnight at 4 °C: anti-Lyve1 antibody (5 μg/ml, mouse, Abcam, ab33682); recombinant anti-mannose receptor antibody [ EPR25215 –277] (1:50, rabbit, Abcam, ab300621); MBP antibody (F-6) (1:50, mouse, SANTA CRUZ Biotechnology, sc-271524); NF-H recombinant SuperclonalTM antibody (2 μg/ml, rabbit, Thermo Fisher, 711,025); PDGFRα antibody (5 μg/ml, goat, RD systems, AF1062); and lectin (594, vector labs, DL-1207–5).

Techniques: Immunofluorescence, Staining

Journal: Development (Cambridge, England)

Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish

doi: 10.1242/dev.204395

Figure Lengend Snippet: Dynamic expression of Nr4a1 after cardiac injury. (A) Expression pattern of nr4a1 at different stages post-injury. The fold change is calculated relative to the expression in uninjured wild-type (wt) hearts. (B) The schematic for generating TgKI(nr4a1:eGFP) . (C) Detection of nr4a1:eGFP fusion mRNA. The 518 bp band indicates correct nr4a1:eGFP fusion cDNA. (D) Western blot analysis of TgKI(nr4a1:eGFP) to identify the fusion Nr4a1:eGPF protein (88kD). (E-H) Expression of Nr4a1, as revealed by GFP antibody staining, in 2 and 7 dpci hearts. (I) IB4 staining to localize Nr4a1 expression in macrophages within 7 dpci hearts. (J) Anti-Mpx antibody staining to localize Nr4a1 expression in neutrophils within 2 dpci hearts. P -values<0.05 were considered statistically significant (two-tailed unpaired t -test). Scale bars: 60 µm.

Article Snippet: For IB4 staining to mark the macrophage (1:100, Griffonia Simplicifolia Lectin I Isolectin B4, Vector Laboratories, DL-1207-.5), the IB4 dye was added in the blocking buffer along with secondary antibody during immunostaining of the sections.

Techniques: Expressing, Western Blot, Staining, Two Tailed Test

Journal: Development (Cambridge, England)

Article Title: Nr4a1 modulates inflammation and heart regeneration in zebrafish

doi: 10.1242/dev.204395

Figure Lengend Snippet: Abnormal accumulation of inflammatory macrophages in nr4a1 mutant. (A-C) Left: distribution of inflammatory macrophages in uninjured wild-type (WT) hearts and WT hearts at 7 and 14 dpci. Right: distribution of inflammatory macrophages in uninjured nr4a1 mutant hearts and nr4a1 mutant hearts at 7 and 14 dpci. Boxed regions in A-C show the approximate positions for quantification, and position of magnified images. Dashed lines show the wounded area. (D,E) Quantification of inflammatory macrophages in the unit area at 7 and 14 dpci. (F-H) qPCR results of inflammation-related genes tnfa , mpeg1.1 and acod1 . All fold changes are calculated relative to the uninjured WT group in each panel. (I) Schematic showing inflammatory macrophage distribution in WT and nr4a1 mutant. Created in BioRender by Feng, D., 2025. https://BioRender.com/ynn17bt . This figure was sublicensed under CC-BY 4.0 terms. P -values<0.05 were considered statistically significant (two-tailed unpaired t -test, D,E; two-way ANOVA with Sidak test for multiple comparison correction and two-tailed unpaired t -test, F-H). Points in D and E show the individual samples. Data are mean±s.e.m. Scale bars: 275 µm.

Article Snippet: For IB4 staining to mark the macrophage (1:100, Griffonia Simplicifolia Lectin I Isolectin B4, Vector Laboratories, DL-1207-.5), the IB4 dye was added in the blocking buffer along with secondary antibody during immunostaining of the sections.

Techniques: Mutagenesis, Two Tailed Test, Comparison