Journal: PLoS ONE

Article Title: Ovariectomy and Subsequent Treatment with Estrogen Receptor Agonists Tune the Innate Immune System of the Hippocampus in Middle-Aged Female Rats

doi: 10.1371/journal.pone.0088540

Figure Lengend Snippet: Expression of macrophage-associated genes and inhibitory neuronal ligands in the hippocampus of middle-aged female rats.

Article Snippet: Cd68 , Rn01495634_g1 , 8.305.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Ovariectomy and Subsequent Treatment with Estrogen Receptor Agonists Tune the Innate Immune System of the Hippocampus in Middle-Aged Female Rats

doi: 10.1371/journal.pone.0088540

Figure Lengend Snippet: Expression of microglia marker Iba1 (A), Cd68 (B), and M2 marker Mrc1 (C) and Cd163 (D) in the hippocampus of middle-aged (M), middle-aged ovariectomized (M/OVX) and M/OVX rats treated with E2 (M/OVX+E2), LE2 (M/OVX+LE2) and DPN (M/OVX+DPN) was determined by real-time PCR. Error bars show SD of five samples for each group. ANOVA identified statistically significant treatment effects for each gene (p = 0.009 for Iba1, p = 0.014 for Cd68, p<0.001 for M2 markers). Turquoise, black and red asterisks mark statistically significant group differences compared to M, M/OVX and M/OVX+E2 animals, respectively, by Newman-Keuls post hoc test. * corresponds to 0.01

Article Snippet: Cd68 , Rn01495634_g1 , 8.305.

Techniques: Expressing, Marker, Real-time Polymerase Chain Reaction

Journal: Physiological Reports

Article Title: Autologous minced muscle grafts improve endogenous fracture healing and muscle strength after musculoskeletal trauma

doi: 10.14814/phy2.13362

Figure Lengend Snippet: Flow cyotometry antibodies

Article Snippet: CD68 , ED1 , Novus Biologicals , NB600‐985V , 0.001 mg/mL.

Techniques: Concentration Assay

Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Gene transfer corrects acute GM2 gangliosidosis--potential therapeutic contribution of perivascular enzyme flow.

doi: 10.1038/mt.2012.44

Figure Lengend Snippet: Figure 6 Inflammatory response in the presence of residual storage and emerging disease in nontargeted tissues. (a) The intensity and number of cells staining for the microglial marker, CD68, and astrocytic marker, GFAP, correlated with local glycoconjugate abundance (stained magenta with periodic acid-Schiff (PAS) reagent). Displayed are sections from an Sx2-injected Sandhoff (SD) mouse, killed at 696 days and viewed in Supplementary Videos S4 and S5 at 12 and 20 months of age. The ninth cerebellar lobule with few PAS-stained neurons (hash area and inset in left panel) had few cells labeled with CD68 (arrowheads in left panel) and GFAP (arrowhead in left panel). In contrast, the flocculus lobe with many intensely PAS-stained neurons (asterisk area and inset in right panel) had many cells labeled with CD68 (arrowheads, right panel) and GFAP (arrow- heads, right panel). Globular microglia (arrows in left bottom panel) and astrocytes (arrows, right bottom panel) are intensely stained. (b) Abundant periodic acid-Schiff (PAS)-stained glycoconjugate was found in retinal ganglion cell layer (GL) in 4-month untreated SD [SD (UT)] compared with 2-year-old wild type (WT). In S+C-treated 2 year-old SD [SD (T)] staining was found in GL and inner nuclear (INL) layers. (c) SD (T) mouse endothe- lium in regions with low activity of B-hexosaminidase contained PAS-stained glycoconjugates (arrows, top panel), but staining was undetectable in regions with abundant enzyme activity such as those close to injection sites (arrows in lower panel). Bars: 2 mm (from top: first set of panels in a); 200 μm (insets in a); 100 μm (c, and from top: second, third, and fourth sets of panels in a); 50 μm (b).

Article Snippet: Immunohistological staining was performed with mouse monoclonal anti-N-acetyl-GM2 immunoglobulin M (MK1-16; Seikagaku, Tokyo, Japan; 1/100), rat anti-mouse CD68 (MCA1957; Serotec, Oxford, UK; 1/50), goat anti-GFAP (N-18, Santa Cruz Biotechnology, Calne, UK; 1/50) as primary antibodies.

Techniques: Staining, Marker, Injection, Labeling, Activity 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: 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

Journal: International Journal of Molecular Sciences

Article Title: Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol

doi: 10.3390/ijms23010223

Figure Lengend Snippet: Secondary antibodies, chromogens, fluorophores and metals. Except DAPI, the fluorophores not directly coupled to antibodies are linked to streptavidin for binding of biotinylated antibodies. Abbreviations: RTU: ready to use, AP: alkaline phosphatase, HRP: horseradish peroxidase. Tb: Terbium, Er: Erbium.

Article Snippet: , 159Tb (conjugated to CD68 clone KP1) , , - , Fluidigm (SKU 201300).

Techniques: Binding Assay