Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 2 High SERPINE1 expression in GC cells promotes macrophage M2 polarization. tSNE visualization of nine single-cell clusters partitioned by unsu pervised cluster analysis, SERPINE1 expression of each single-cell, and SERPINE1 expression abundance of different single-cell clusters in the GSE134520 (A–C) and GSE167297 (D–F) datasets. (G) Flow cytometry analysis of the proportion of CD68+CD206+ macrophages in a Transwell co-culture system, with MKN45 and AGS cells overexpressing (oe_SERPINE1) or silencing SERPINE1 (shRNA#3 or sh_SERPINE1#3) in the upper chamber, and THP1 cells treated with PMA in the lower chamber. (H) Immunofluorescence staining of xenograft tumor tissues. Comparison of the proportion of M1 or M2 macrophage infiltra tion. Green indicates F4/80. Red indicates iNOS or Arg1 expression

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, shRNA, Immunofluorescence, Staining, Comparison

Journal: Journal of experimental & clinical cancer research : CR

Article Title: Gastric cancer-derived exosomal let-7 g-5p mediated by SERPINE1 promotes macrophage M2 polarization and gastric cancer progression.

doi: 10.1186/s13046-024-03269-4

Figure Lengend Snippet: Fig. 5 SERPINE1-mediated gastric cancer-derived exosomes facilitate the polarization of THP1 cells into M2 macrophages. (A) Schematic representation of the extraction and identification of exosomes and the induction of macrophage polarization. Transmission electron microscopy (B), nanoparticle tracking analysis (C), and western blotting (D) were used to identify the morphology, particle size, and markers of exosomes. (E) Confocal laser scanning microscopy detected Dil-labeled exosomes (red) internalized by DAPI-labeled macrophages (blue). (F–G) Immunofluorescence analysis of the proportion of CD206+ cells in THP1 cells treated with exosomes. (H–I) Flow cytometry analysis of the proportion of CD68+CD206+ cells in THP1 cells treated with exosomes. (J–K) qRT-PCR analysis of M1 markers (iNOS and TNF-α) and M2 markers (TGF-β, IL-10, and Arg-1) in THP1 cells treated with exosomes. (L–N) Transwell migration and invasion assays of GC cells (upper chamber) co-cultured with macrophages (lower chamber) ingesting exosomes

Article Snippet: THP-1 cells were differentiated into macrophages using 150 ng/mL phorbol 12-myristate 13-acetate (PMA, Sigma) for 24 h and subsequently co-cultured with cancer-derived exosomes or GC cells in 6-well plates with 0.4-μm membranes for 72 h. Harvested macrophages were converted into single-cell suspensions, stained with Elab Fluor 488 anti-human CD68 (Mouse, 1:20, ElabScience) and APC anti-human CD206 (Mouse, 1:20, ElabScience) antibodies, and analyzed for CD68+CD206+ populations by flow cytometry (Accuri C6, BD).

Techniques: Derivative Assay, Extraction, Transmission Assay, Electron Microscopy, Western Blot, Confocal Laser Scanning Microscopy, Labeling, Immunofluorescence, Flow Cytometry, Quantitative RT-PCR, Migration, Cell Culture

Journal: Antioxidants

Article Title: Genipin Attenuates Diabetic Cognitive Impairment by Reducing Lipid Accumulation and Promoting Mitochondrial Fusion via FABP4/Mfn1 Signaling in Microglia

doi: 10.3390/antiox12010074

Figure Lengend Snippet: GEN shifted microglia polarization and inhibited lipid accumulation in LPS/HG/PA-induced HMC3 cells. The HMC3 cells were treated with GEN (5, 10, 20 μM) for 4 h and then stimulated with LPS/HG/PA for 12 h. The supernatant concentrations of TNF-α ( A ), IL-6 ( B ), IL-4 ( C ) and IL-10 ( D ) were examined by ELISA (n = 4). The mRNA expressions of iNOS ( E ), CCL2 ( F ), ARG1 ( G ) and YM1 ( H ) in cells were assessed by PCR (n = 4). The population of CD68-positive and CD206-positive cells were measured by flow cytometry ( I ). The expressions of FABP4, p-NF-κB and NF-κB were detected by Western blot ( J – L ) (n = 3). The nucleus translocation of NF-κB was visualized by immunofluorescence staining under laser confocal microscope. The scale bar equaled 5 μm ( M ). The lipid accumulation was observed by oil red O staining. The scale bar equaled 50 μm ( N ). The analysis of oil red O staining ( O ). The mRNA expressions of fatty acid β-oxidation genes including ACOX1 , ACAA2 and ECHS1 were measured by PCR ( P ). The mRNA expressions of fatty acid uptake genes including SLC27A1 and PPARα were measured by PCR ( Q ). The mRNA expressions of fatty acid synthesis genes including FASN and ACLY were measured by PCR ( R ). The results are expressed as means ± SDs. ### p < 0.001 compared with control group. * p < 0.05, ** p < 0.01 compared with LPS/HG/PA group or the other group. ns means not significant.

Article Snippet: Mfn2 (#9482S), Drp1 (#8570S), p-Nuclear Factor Kappa B (NF-κB) (#3033), NF-κB (#8242), CD68 (#91882), p47phox (#4312), β-actin (#4970), Na, K-ATPase (#3010), ubiquitin (#3936), Flag (#8146), Flag (#14793), HA (#3724), Myc (#2276), V5 (#80076), mouse anti-rabbit IgG (Conformation Specific, #5127), rabbit anti-mouse IgG (Light Chain Specific, #58802) and mouse anti-rabbit IgG (Light-Chain Specific, #93702) antibodies were provided by Cell Signaling Technology (Danvers, MA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Western Blot, Translocation Assay, Immunofluorescence, Staining, Microscopy, Control

Journal: The Journal of Neuroscience

Article Title: Microglial mTOR is Neuronal Protective and Antiepileptogenic in the Pilocarpine Model of Temporal Lobe Epilepsy

doi: 10.1523/JNEUROSCI.2754-19.2020

Figure Lengend Snippet: Key resources, related to experimental procedures

Article Snippet: Rat anti-mouse CD68 , Bio-Rad , Catalog #MCA1957; RRID: AB_322219.

Techniques: Recombinant, In Situ, cDNA Synthesis, SYBR Green Assay, Software

Journal: The Journal of Neuroscience

Article Title: Microglial mTOR is Neuronal Protective and Antiepileptogenic in the Pilocarpine Model of Temporal Lobe Epilepsy

doi: 10.1523/JNEUROSCI.2754-19.2020

Figure Lengend Snippet: Activation of mTOR in microglia following SE. A, Confocal images acquired from CTX, hippocampus CA1 and CA3, and the DG. Scale bar, 20 µm. B, C, Quantification of p-S6/Iba1 double-positive microglial cells (B) and CD68/Iba1 double-positive microglial cells (C) in sham-treated (n = 4; 2 males and 2 females), SE-3d (n = 5; 2 males and 3 females), and SE-3d+rapamycin (n = 4; 2 males and 2 females) mice. Data are presented as the mean ± SEM. Sample comparison was made among three groups (sham, SE, and SE + rapamycin groups) and analyzed with one-way ANOVA (P1), followed by Tukey's multiple-comparisons test (P2, comparison between control and SE; P3, comparison between SE and SE + rapamycin).

Article Snippet: Rat anti-mouse CD68 , Bio-Rad , Catalog #MCA1957; RRID: AB_322219.

Techniques: Activation Assay, Comparison, Control

Journal: The Journal of Neuroscience

Article Title: Microglial mTOR is Neuronal Protective and Antiepileptogenic in the Pilocarpine Model of Temporal Lobe Epilepsy

doi: 10.1523/JNEUROSCI.2754-19.2020

Figure Lengend Snippet: Microglial proliferation and activation in control and mTORCx3cr1-creCKO mice following SE. A, Full montage of confocal images showing acute injury (FJB+, green), microglial density (Iba1+, green), and CD68 expression (CD68+, red) in the hippocampus of control and mTORCx3cr1-creCKO mice either sham-treated or 1–7 d post-SE. Scale bar, 200 µm. B, Representative confocal images acquired from the CA1 of control and mTORCx3cr1-creCKO mice either sham treated or 1–7 d post-SE. Scale bar, 20 µm. C, Representative confocal images showing Ki67 expression (Ki67+, red) in the CA1 of control and mTORCx3cr1-creCKO mice either sham treated or 1–7 d post-SE. Scale bar, 20 µm. D, Average doses of pilocarpine that induced stage 4/5 seizures in control and mTORCx3cr1-creCKO mice. E, F, Quantification of Iba1+ microglia and CD68 immunofluorescence intensity in control and mTORCx3cr1-creCKO mice either sham-treated or 1-7 d post-SE. Sample sizes in control groups: sham (n = 6; 3 males and 3 females), SE-1d (n = 6; 3 males and 3 females), SE-3d (n = 6; 3 males and 3 females), and SE-7d (n = 6; 3 males and 3 females); and in the mTORCx3cr1-creCKO groups: sham (n = 4; 2 males and 2 females), SE-1d (n = 5; 2 males and 3 females), SE3d (n = 6; 3 males and 3 females), and SE-7d (n = 4; 2 males and 2 females). G, Quantification of Ki67/Iba1 double-positive microglial cells in control and mTORCx3cr1-creCKO mice either sham treated or 1–7 d post-SE. Sample sizes in control groups: sham (n = 6; 3 males and 3 females), SE-1d (n = 6; 3 males and 3 females), SE-3d (n = 6; 3 males and 3 females), and SE-7d (n = 6; 3 males and 3 females); and in the mTORCx3cr1-creCKO groups: sham (n = 4; 2 males and 2 females), SE-1d (n = 5; 2 males and 3 females), SE3d (n = 6; 3 males and 3 females), and SE-7d (n = 4; 2 males and 2 females). Data are presented as the mean ± SEM A two-tailed unpaired t test was used for the comparison between control and mTORCx3cr1-creCKO mice.

Article Snippet: Rat anti-mouse CD68 , Bio-Rad , Catalog #MCA1957; RRID: AB_322219.

Techniques: Activation Assay, Control, Expressing, Immunofluorescence, Two Tailed Test, Comparison

Journal: IBRO neuroscience reports

Article Title: IL-33 relieves nerve injury by mediating microglial polarization in neuromyelitis optica spectrum disorders via the IL-33/ST2 pathway.

doi: 10.1016/j.ibneur.2024.07.008

Figure Lengend Snippet: Fig. 5. Microglial cells were activated and transformed to the M2 phenotype after treatment with IL-33. (a) Western blot images and the quantification of CD206 and CD40 levels after treatment with IL-33 or NMOD serum. (b) BV2 cells were labeled with Iba-1 and CD68 for immunofluorescence. (c) Expression levels of PSD95 were observed by immunofluorescence after treatment with IL-33 or NMOSD serum. One-way analysis of variance was used for statistical analyses. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: For protein immunoreactivity in cells, the primary cortical neurons or BV2 cells were fixed with 4 % paraformaldehyde for 30 min at RT and treated with 0.5 % Triton for 10 min. After being washed with PBS, the cells were blocked with 5 % bovine serum albumin (BSA) for 1 h and then incubated with a rabbit antiPSD95 antibody (1:100 dilution, PA2295, Boster), anti-CD68 antibody (1:200 dilution, 28058–1-AP, Proteintech), anti-IBA1 antibody (1:100 dilution, CY7217, Abways), or anti-ST2 antibody (1:200 dilution, PRS3363, Sigma) at 4 ◦C overnight.

Techniques: Transformation Assay, Western Blot, Labeling, Immunofluorescence, Expressing

Journal: Theranostics

Article Title: Therapeutic effects of a novel electrode for transcranial direct current stimulation in ischemic stroke mice

doi: 10.7150/thno.90779

Figure Lengend Snippet: Comparison of therapeutic effects between conventional and new HD-tDCS applications in stroke. (A) Schematic diagram of the new HD electrode and 3D simulation of the HD-tDCS application. Higher relative electric potential and enhanced spatial focality were predicted in the motor cortex using the new HD-tDCS application compared with those using the conventional HD-tDCS application. (B) Timeline for HD-tDCS application and behavioral tests. Both HD-tDCS applications were used to treat the contralesional motor cortex once daily for 5 days. (C) Corner test: The percentage of left turns was significantly decreased in the MCAO+tDCS-FN group compared with that in the MCAO and MCAO+tDCS-F groups. (D) Pole test: The total time for the MCAO+tDCS-FN group was significantly decreased compared with that of the MCAO group. (E) Bar charts and performance images of the open-field test. The time in the center zone was significantly increased in the MCAO+tDCS-FN group compared with that in the MCAO and MCAO+tDCS-F groups. (F) Schematic subject images, bar charts, and performance images for the novel object recognition test. The recognition index was significantly enhanced in the MCAO+tDCS-FN group compared with that in the MCAO group. (G) Schematic diagram, bar charts, and schematic recording configuration of the movement during the fear conditioning. The percentage of freezing in the CS was significantly enhanced in the MCAO+tDCS-FN group compared with that in the MCAO+tDCS-F group. (H) Representative brain images (-0.1 mm from the bregma) and quantification of the relative infarct volume. Borders to the healthy tissue are indicated with a dot line. The infarct volume in 40 brain sections from 1.1 mm to -1.9 mm from the bregma was significantly reduced in the MCAO+tDCS-FN group compared with that in the MCAO+tDCS-F group. (I) Photomicrograph and bar chart showing the number of Iba1- and CD68-positive cells in the peri-infarct region. The number of Iba1-positive cells was significantly reduced in the MCAO+tDCS-FN group compared with that in the MCAO+tDCS-F group. All behavioral, morphological, and immunofluorescence analyses (n=5/group). Data are presented as means±SEMs. * P <0.05 and *** P <0.001 vs. each group using one-way analysis of variance with Tukey's test. IL, ipsilesional site; CL, contralesional site; MCAO, middle cerebral artery occlusion; HD-tDCS, high-definition transcranial direct current stimulation; F, conventional electrode; FN, new needle-conjugated electrode; BL, baseline; CS, cue stimulation; TI, trace interval. Magnification: ×400; scale bar=100 μm.

Article Snippet: They were then incubated overnight with the following primary antibodies (diluted in antibody dilution buffer, 1× PBS/1% bovine serum albumin/0.3% Triton X-100) at 4°C: ionized calcium binding adaptor molecule 1 (Iba1) (1:500, 019-19741; Wako Chemicals, Richmond, VA), CD68 (1:500, sc-135872, Santa Cruz Biotechnology), NeuN (1:500, MAB377, Millipore, Billerica, MA, USA), SREBP1 (1:100, NB100-74542, Novus Biologicals), pNFkB1 (1:100; PA5-37658, Invitrogen), and c-Cas3 (1:50; 9661, Cell Signaling Technology).

Techniques: Comparison, Immunofluorescence

Journal: Biochemistry and Biophysics Reports

Article Title: Oxymatrine blocks the NLRP3 inflammasome pathway, partly downregulating the inflammatory responses of M1 macrophages differentiated from THP-1 monocytes ☆

doi: 10.1016/j.bbrep.2023.101482

Figure Lengend Snippet: Morphological images of cells in different states and identification of M0 and M1 macrophages. (A) Morphology of THP-1 cells, M0 macrophages, and M1 macrophages. Scale bar: 200 μm. THP-1 monocytes were round, translucent, and suspended; M0 macrophages were clustered and adherent; M1 macrophages were adherent and can extend pseudopodias. (B) Immunofluorescence staining on M0 cells. Scale bar: 50 μm. Colocalization of CD68 (red) and DAPI (blue). (C) Immunofluorescence staining of M1 cells. Scale bar: 50 μm. Colocalization of CD86 (red) and DAPI (blue). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For immunofluorescence staining, the cells were fixed with 4% paraformaldehyde (PFA) (Biosharp, Anhui, China) and incubated with diluted rabbit primary antibody against CD68 (1:300) (Boster, Wuhan, China) overnight at 4 °C.

Techniques: Immunofluorescence, Staining

Journal: International Journal of Molecular Medicine

Article Title: Effects of ketanserin on experimental colitis in mice and macrophage function

doi: 10.3892/ijmm.2016.2486

Figure Lengend Snippet: 5-Hydroxytryptamine-2A receptor (5-HT 2A R) expression is upregulated in the colons of patients with inflammatory bowel disease (IBD) and in mice with dextran sodium sulfate (DSS)-induced colitis, and is specifically enhanced in macrophages. (A) Relative mRNA expression of 5-HT 2A R in healthy subjects and patients wtih IBD estimated by RT-qPCR. (B) Representative images of immunofluorescence staining of colonic mucosa samples of healthy subjects and in patients with IBD showing 5-HT 2A R + cells (in green) and CD68 + cells (in red). Co-localization of 5-HT 2A R with the macrophage marker, CD68 + , is shown in the merged images. 5-HT 2A R expression in mice with DSS-induced colitis estimated by (C) RT-qPCR and (D) western blot analysis. Representative images of immunofluorescence staining of 5-HT 2A R + cells (in green) and CD68 + cells (in red) in the colonic mucosa samples of mice treated with or without DSS. (E) Co-localization of 5-HT 2A R with CD68 is shown in the merged images.

Article Snippet: Mouse anti-human CD68 antibody (MCA5709; mouse anti-human CD68, monoclonal antibody; AbD Serotec, Kidlington, UK; 1:500) (overnight at 4°C) was subsequently used to detect the macrophages.

Techniques: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Marker, Western Blot

Journal: Cell Reports Medicine

Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

doi: 10.1016/j.xcrm.2024.101397

Figure Lengend Snippet: Microbial distribution in human solid tumors is spatially heterogeneous and coupled with CD8 + T cell infiltration (A) Immunofluorescence (IF) images of CK19 (green), LPS (red), and nuclei (DAPI, blue) (top row) and heatmaps of LPS (bottom row). Scale bars, 2 mm. (B and C) IF images of LPS-poor (left) and LPS-rich (right) regions. Scale bars, 200 μm. (D) Spatial variance of LPS. (E) Distribution of LPS in CK19 + tumor cells (yellow) and stromal cells (purple). (F) Study design. (G) Quantification of LPS. CD20 (n = 56 for CD20-poor and n = 11 for CD20-enriched nests pooled from 7 PDAC specimens), CD68 (n = 43 for CD68-poor and n = 28 for CD68-enriched nests pooled from 8 PDAC specimens), and FOXP3 (n = 49 for FOXP3-poor and n = 16 for FOXP3-enriched nests pooled from 9 PDAC specimens). Scale bars, 100 μm. (H) IF images of CD8 (yellow), tumor cells (green, CK19), LPS (red), and nuclei (blue, DAPI). Scale bars, 100 μm. (I) Quantification of LPS in PDAC tumor nests (left; n = 110 for CD8-poor nests and n = 39 for CD8-enriched nests pooled from 10 PDAC specimens). (J) Quantification of LPS in LUAC tumor nests (right; n = 100 for CD8-poor nests and n = 50 for CD8-enriched nests pooled from 10 LUAC specimens). For (D) and (E), n = 9 for PDAC and n = 9 for LUAC. Statistical significance was calculated using a two-tailed Mann-Whitney test (E, G, I, and J). Data are represented as violin plots (center line, median; top and bottom lines, upper and lower quartiles) and scatterplots (mean ± SD). PDAC, pancreatic ductal adenocarcinoma; LUAC, lung adenocarcinoma; NS, not significant.

Article Snippet: Rabbit monoclonal anti-mouse CD68 (E3O7V) , Cell Signaling Technology , 29176SF.

Techniques: Immunofluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

doi: 10.1016/j.xcrm.2024.101397

Figure Lengend Snippet: T cell and microbial co-localization define distinct cellular neighborhoods in human pancreatic cancer (A) Study design for (B)–(D) (n = 185 for cold nests and n = 77 for hot nests pooled from 16 specimens). (B) Images of CD8 (yellow), CD68 (brown), tumor epithelium (teal, CK19), FOXP3 (purple), and nuclei (blue, hematoxylin). Scale bars, 1 mm (top) and 50 μm (bottom). (C) Quantification of CD8 + (left), FOXP3 + (middle), and CD68 + (right) cells. (D) Ratio of CD8 + cells to FOXP3 + cells. (E) Study design for (F) and (H)–(J). (F) Principal-component analysis (n = 5 for hot and cold tumor nests from 1 specimen). (G) 16S rRNA levels (n = 14 for cold and hot tumor nests from 3 patient specimens). (H and I) Heatmap (H) and enrichment score (I) for the relative expression of genes associated with response to bacterium (GO: 0009617) in hot and cold stroma. (J) Volcano plot of DEGs. (K) Multiplex immunohistochemistry images of CD8 (yellow), PIGR (purple), and nuclei (blue, hematoxylin). (L) Quantification of PIGR (n = 49 for cold nests and n = 46 for hot nests pooled from 7 specimens). (M) Multiplex immunohistochemistry images of CD8 (teal), CD20 (yellow), CD74 (purple), and nuclei (blue, hematoxylin). (N) Quantification of CD74 (n = 71 for cold nests and n = 45 for hot nests pooled from 10 specimens). (O) Summary of cell markers and proteins. For (K) and (M), scale bars, 2 mm (left) and 50 μm (right), and dashed lines indicate tumor epithelium. Statistical significance was calculated using a two-tailed Mann-Whitney test. Data are represented as violin plots (center line, median; top and bottom lines, upper and lower quartiles) and scatterplots (mean + SD or mean). PDAC, pancreatic ductal adenocarcinoma.

Article Snippet: Rabbit monoclonal anti-mouse CD68 (E3O7V) , Cell Signaling Technology , 29176SF.

Techniques: Expressing, Multiplex Assay, Immunohistochemistry, Two Tailed Test, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Multimodal immune phenotyping reveals microbial-T cell interactions that shape pancreatic cancer

doi: 10.1016/j.xcrm.2024.101397

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-mouse CD68 (E3O7V) , Cell Signaling Technology , 29176SF.

Techniques: Recombinant, Sequencing, Software

Journal: Molecular brain

Article Title: Key mechanisms of angiogenesis in the infarct core: association of macrophage infiltration with venogenesis.

doi: 10.1186/s13041-025-01182-1

Figure Lengend Snippet: Fig. 5 Phagocytosis was associated with venogenesis in the infarct core. (A) Transmission electron microscopy images of the infarct core on day 10. (a1) Foamy macrophages in the infarct region. (a2) Magnification of lipid-rich foamy macrophages. (a3) Macrophage engulf cell debris. (a4) A microvessel was spatially close to macrophages. ly, lysosome; lp, lipid droplets; N, nucleus; MCH, macrophage; ECs, endothelial cells; BL, basal lamina; P, pericytes. (B) GIAO staining revealed lipid-rich foamy macrophages and newly formed venules in the infarct core. (C) The top 20 upregulated phagocytosis-associated genes (fold change ≥ 2 and p < 0.05) in the infarct core (n = 5). (D) mRNA expression of macrophage markers of different phenotypes on day 10. n = 4. *p < 0.05 vs. control. ※p < 0.05 vs. peri. (E) Immunofluorescence images of CD68-, Arg1-, and Cd206-labeled macrophages in the infarct core on days 3 and 7. Scale bar, 50 μm. (F) ALP staining of microvessels and EPCs in the cortex. (f1) Arterioles in the normal cortex. (f2) and (f3) ALP-labeled EPCs in the infarct region of control mice. (f4) ~ (f6) ALP-labeled EPCs in mice with macrophage depletion (mac-/-). EPCs proliferate and form microvessels from the outer edge of the infarct cortex. (G) Representative CD31-labeled microvessels in the infarct cores of control and mac-/- mice. (H) and (I) Quantitative analysis revealed increased vessel density (ALP, p < 0.0001; CD31, p = 0.006) in mac-/- mice compared with control mice. *p < 0.05

Article Snippet: The following primary antibodies were used to incubate the brain sections overnight at 4 °C: sheep polyclonal anti-BrdU (Abcam), purified rat antimouse CD31 (BD PharmingenTM), 488-conjugated Arginase-1 polyclonal (Proteintech), 488 rabbit anti-mouse CD68 (ABclonal Technology), and CD206 monoclonal (MR5D3) (Invitrogen) antibodies.

Techniques: Transmission Assay, Electron Microscopy, Staining, Expressing, Control, Immunofluorescence, Labeling