Journal: Biology of reproduction

Article Title: Distribution of immune cells in the epididymis of the aging Brown Norway rat is segment-specific and related to the luminal content.

doi: 10.1095/biolreprod61.3.705

Figure Lengend Snippet: FIG. 1. Light micrographs showing sec- tions of the epididymis of the Brown Nor- way rat stained with an antibody for monocytes-macrophages (ED1). A–C) Cor- pus, D–F) distal cauda. A, D) 3 mo, B, E) 18 mo with numerous spermatozoa in the lumen, C, F) 18 mo with occasional sper- matozoa in the lumen. lu, Lumen; it, inter- tubular space; p, principal cells; b, basal cells; c, clear cells; bm, basement mem- brane. Arrows, ED1-positive cells; aster- isks, halo cells. Scale bar, A–F 5 64 mm.

Article Snippet: Mouse anti-rat ED1 (Serotec USA, Washington, DC) was used at a dilution 1/100; this antibody recognizes a cytoplasmic antigen in monocytes and most macrophages.

Techniques: Staining

Journal: Biology of reproduction

Article Title: Distribution of immune cells in the epididymis of the aging Brown Norway rat is segment-specific and related to the luminal content.

doi: 10.1095/biolreprod61.3.705

Figure Lengend Snippet: FIG. 5. Light micrographs showing sec- tions of the corpus of the epididymis of Brown Norway rats. A, C) Three mo; B, D) 18 mo. Section stained with an antibody for A and B GST Yf and C and D ED1. Clear arrows, basal cells; dark arrows, ED1-positive cells; other labels as in Fig- ure 1. Scale bar, A–D 5 64 mm.

Article Snippet: Mouse anti-rat ED1 (Serotec USA, Washington, DC) was used at a dilution 1/100; this antibody recognizes a cytoplasmic antigen in monocytes and most macrophages.

Techniques: Staining

Journal: Stem cell research & therapy

Article Title: Subretinal microglia support donor photoreceptor survival in rd1 mice.

doi: 10.1186/s13287-024-04052-0

Figure Lengend Snippet: Fig. 2 Macrophage and microglia detection after transplantation. A. In rd1 mice, macrophages (CD68+) were observed in the choroid when donor cells either survived (left) or died (middle). In wild-type mice, dead donor cells were engulfed by macrophages (right). Scale bar = 10 μm. B. In trans-scleral injection group, while microglial cells surrounded surviving donor cells in rd1 mice (i), they could hardly be detected around dead cell debris (ii). In wild-type mice, microglia/macrophages engulfed dead donor cells (iii). Following trans-vitreous injection, both dead (arrows) and surviving donor cells (arrowheads) were found in the subretinal space. While macrophages/microglia engulfed the dead cells (arrows), microglia were not close to surviving cells (iv). C. A Comparison of chemokines expression between wild-type and rd1 mice showed a significant increase in the mRNA levels of ccl2, ccl3 and cxcl2 in rd1 mice. D. The comparison of cytokine levels between wild-type and rd1 mice showed no significant difference. E. No significant differences were found in oxidative stress factors between wild-type and rd1 mice. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The following primary antibodies were used in the study: CD68 (1:400,#97778, Cell Signaling Technology), Iba1 (1:200, #019-19741, Wako), Galectin-3 (1:200, #sc-53127, Santa Cruz), F4/80(1:400, #71299, Cell Signaling Technology), CD11b (1:200, 101,209, Biolegend).

Techniques: Transplantation Assay, Injection, Comparison, Expressing

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: PLoS ONE

Article Title: Cholesterol Diet Withdrawal Leads to an Initial Plaque Instability and Subsequent Regression of Accelerated Iliac Artery Atherosclerosis in Rabbits

doi: 10.1371/journal.pone.0077037

Figure Lengend Snippet: Panel (A) Representative images of hematoxylin and eosin stained sections of all groups (Scale bar = 500 µm). Panel (B) Representative images of Oil Red O stained cryostat sections of all groups (Scale bar = 50 µm). Panel (C) Immunohistochemical staining showing CD68 positive cells in respective groups. (Scale bar = 50 µm) (D) Quantitative analysis of percentage lipid area (lumen area included) in respective groups (E) Quantitative analysis of CD68 positive labelling in respective groups. **p<0.01 and ***p<0.001 vs normal; ## p<0.01 and ### p<0.001 vs Baseline. (* indicates lumen, arrow heads indicate positive staining)

Article Snippet: Paraffin-embedded consecutive sections were stained with the following antibodies: Mouse monoclonal anti rabbit CD68 (RAM11, Dako, USA), alpha smooth muscle actin (1A4, Sigma, USA) and MMP-9 (56-2A4, Calbiochem, USA).

Techniques: Staining, Immunohistochemical staining

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: White matter microglia/macrophages are activated in the dorsal funiculi of male Plp1 mutant mice. Cervical spinal cord sections from ( A ) mildly-affected P16 rsh ( B ) severely-affected P16 msd ( C ) subclinical P25 Plp1 #66 heterozygous ( D ) moderately-affected P25 Plp1 #66 homozygous mice labeled with antibodies against Iba-1 (red) and CD68 (green). Many Iba-1 positive microglia/macrophages with swollen cell bodies and short processes are labeled with anti-CD68 antibodies in rsh and msd mice, which is indicative of activated cells. The insets in these panels show the canonical activated morphology revealed with anti-Iba-1 antibodies and strong CD68 staining. In contrast, there is little evidence for broad CD68 labeling in Plp1 #66 heterozygous mice in panel C, which is consistent with previous data that immune cells are rarely activated . Activation of microglia/macrophages is apparent for Plp1 #66 homozygous mice in panel D, which exhibit an intermediate level of CD68 labeling. Scale bar: 95 μm, insets 15 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Mutagenesis, Labeling, Staining, Activation Assay

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Proportion of cervical spinal cord Iba-1 + microglia/macrophages in dorsal column, ventral column and lateral funiculus that are activated (CD68 + ).

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques:

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Oligodendrocytes in gray matter from P16 rsh and msd mice do not undergo a UPR and microglia/macrophages exhibit resting morphology. ( A – C ) Iba1 (red) and CD68 (green) antibody labeling in dorsal spinal cord white matter tracts of P16 wild type ( A ) rsh ( B ) and msd ( C ) mice. Although the morphology and CD68 staining is characteristic of resting microglia/macrophages in wild type mice (white arrowheads), these cells are activated in the Plp1 mutants. ( D – F ) In contrast to white matter regions, microglia/macrophages have a resting state phenotype in the adjacent substantia gelatinosa (gray matter) from wild type and mutant mice (black arrowheads). ( G – I ) A major difference between these regions in rsh and msd mice is the relative abundance of oligodendrocytes undergoing an unfolded protein response (UPR), which is evident by the large number of CHOP + cells (green) in white matter (above the dotted line) compared to gray matter (below). We have previously shown that 100% of CHOP + cells in these mutants are oligodendrocytes . Dotted lines mark the white/gray matter boundary. Scale bar in I: 100 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Antibody Labeling, Staining, Mutagenesis

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Activation of rsh and msd microglia/macrophages in optic nerve at P16. ( A ) Longitudinal section from wild type mouse reveals Iba-1 + microglia/macrophages (red) with non-activated morphology (arrowheads). Most of these cells do not express CD68 or express the protein at low levels (green). ( B ) DAPI staining showing the nuclei of the microglia/macrophages in ( A ). ( C , E ) Iba-1 + microglia/macrophages from rsh ( C ) and msd ( E ) mice exhibit an activated morphology with enlarged cell bodies and thickened processes. Most of these cells express CD68 at high levels, which is localized to perinuclear regions. ( D , F ) DAPI staining of the fields in ( C , E ). Scale bar in F: 30 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Activation Assay, Staining

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Expression fold changes of other non-chromosome 17 interferon-induced genes in microarray data for rsh and msd mice.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Expressing, Microarray