Journal: Journal of dairy science

Article Title: M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

doi: 10.3168/jds.2024-25500

Figure Lengend Snippet: Figure 3. Hepatic macrophage counts and associated genetic changes in healthy cows and dairy cows with SCK. (A) Hepatic protein levels of Cluster of Differentiation 68 (CD68) in healthy (n = 6) and SCK (n = 6) cows. (B) Quantification of hepatic CD68 protein levels. (C) Relative hepatic mRNA levels of CD68. (D) Relative mRNA expression levels of (i) C-type lectin domain family 4 member F (CLEC4F) and (ii) small inducible cytokine subfamily B (Cys-X-Cys), member 10 (CXCL10) in the liver of healthy (n = 6) and SCK (n = 6) cows. (E) Levels of (i) CXCL1 and (ii) CCL2 in the liver of healthy (n = 6) and SCK (n = 6) cows. (F) Representative images (scale bar = 50 μm) of immunofluorescence for CD68 (pink) and DAPI (sky blue) in bovine liver. (G) Quantification of MFI of CD68 in the liver of healthy (n = 6) and SCK (n = 6) cows. Data are presented as mean ± SEM. Statistical differences were assessed by t-test.

Article Snippet: The primary antibodies included: rabbit anti-NFκB polyclonal antibody (1:100; bs-0465R, Bioss), rabbit anti- CD68 polyclonal antibody (1:100; bs-1432R, Bioss), mouse anti- CD68 monoclonal antibody (1:100; NB600–985, NOVUS), rabbit anti-CD86 polyclonal antibody (1:100; bs-1035R, Bioss), rabbit anti-CD206 Recombinant antibody (1:100; 81525–1-RR, Proteintech).

Techniques: Expressing, Immunofluorescence

Journal: Journal of dairy science

Article Title: M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

doi: 10.3168/jds.2024-25500

Figure Lengend Snippet: Figure 4. Hepatic macrophages in SCK cows are predominantly M1 polarized. (A) Representative Western blots of inducible nitric oxide sythase (iNOS), CD86, IL-1β and β-actin in the liver of healthy (n = 6) and SCK (n = 6) cows. (B) Quantification of protein levels of (i) iNOS, (ii) CD86 and (iii) IL-1β. (C) Relative mRNA expression levels of iNOS, CD86, IL-1β, IL-6, IL-12b and chemokine (C-C motif) ligand 3 (CCL3) in the liver of healthy (n = 6) and SCK (n = 6) cows. (D) Levels of IL-1β in the liver of healthy (n = 6) and SCK (n = 6) cows. (E) Representative images (scale bar = 20 μm) of immunofluorescence for CD68 (green), CD86 (red) and DAPI (blue) in bovine liver. (F) Quantification of MFI of CD86 in the liver of healthy (n = 6) and SCK (n = 6) cows. (G) Quantification of the number of CD68+/CD86+ positive cells in the liver of healthy (n = 6) and SCK (n = 6) cows. (H) Representative images (scale bar = 50 μm) of immunofluorescence for IL-1β (yellow), iNOS (red) and DAPI (blue) in bovine liver. (I) Quantification of MFI of (i) IL-1β and (ii) iNOS in the liver of healthy (n = 6) and SCK (n = 6) cows. Data are presented as mean ± SEM. Statistical differences were assessed by t-test.

Article Snippet: The primary antibodies included: rabbit anti-NFκB polyclonal antibody (1:100; bs-0465R, Bioss), rabbit anti- CD68 polyclonal antibody (1:100; bs-1432R, Bioss), mouse anti- CD68 monoclonal antibody (1:100; NB600–985, NOVUS), rabbit anti-CD86 polyclonal antibody (1:100; bs-1035R, Bioss), rabbit anti-CD206 Recombinant antibody (1:100; 81525–1-RR, Proteintech).

Techniques: Western Blot, Expressing, Immunofluorescence

Journal: Journal of dairy science

Article Title: M1 polarization of hepatic macrophages in cows with subclinical ketosis is an important cause of liver injury.

doi: 10.3168/jds.2024-25500

Figure Lengend Snippet: Figure 5. Hepatic macrophage M2 polarization decreased in SCK cows. (A) Representative Western blots of CD206, IL-10 and β-actin in the liver of healthy (n = 6) and SCK (n = 6) cows. (B) Quantification of hepatic (i) CD206 and (ii) IL-10 protein levels. (C) Relative hepatic mRNA levels of CD206, IL-10 and arginase 1 (Arg1). (D) Levels of IL-10 in the liver of healthy (n = 6) and SCK (n = 6) cows. (E) Representative images (scale bar = 20 μm) of immunofluorescence for CD68 (green), CD206 (pink) and DAPI (blue) in bovine liver. (F) Quantification of MFI of CD206 in the liver of healthy (n = 6) and SCK (n = 6) cows. (G) Quantification of the number of CD68+/ CD206+ positive cells in the liver of healthy (n = 6) and SCK (n = 6) cows. Data are presented as mean ± SEM. Statistical differences were assessed by t-test.

Article Snippet: The primary antibodies included: rabbit anti-NFκB polyclonal antibody (1:100; bs-0465R, Bioss), rabbit anti- CD68 polyclonal antibody (1:100; bs-1432R, Bioss), mouse anti- CD68 monoclonal antibody (1:100; NB600–985, NOVUS), rabbit anti-CD86 polyclonal antibody (1:100; bs-1035R, Bioss), rabbit anti-CD206 Recombinant antibody (1:100; 81525–1-RR, Proteintech).

Techniques: Western Blot, Immunofluorescence

Journal: PLoS ONE

Article Title: Silver and Gold Nanoparticles Exposure to In Vitro Cultured Retina – Studies on Nanoparticle Internalization, Apoptosis, Oxidative Stress, Glial- and Microglial Activity

doi: 10.1371/journal.pone.0105359

Figure Lengend Snippet: Fluorescent immunostaining: Iba1 (green) and ED1 (red) for detecting microglial cells (DAPI, blue). In the NP-exposed retinas (20 nm Ag (G–I), 80 nm Ag (J–L), 20 nm Au (M–O), 80 nm Au (P–R)) as well as in the control retinas (A–C) the Iba1- and ED–positive cells were located in the IPL, INL and GCL in all groups (A–C). AgNO 3 -exposed retina displayed the same staining pattern, but with a stronger intensity especially for the ED1-staining (D–F). Graph shows numbers of microglial cells and data is given as mean ±SD (n = 4 explants/group). * p <0.05 compared to control. GCL = ganglion cell layer, INL = inner nuclear layer, IPL = inner plexiform layer, ONL = outer nuclear layer, OPL = outer plexiform layer. Scale bar equals 200 µm.

Article Snippet: Sections were then incubated with primary antibodies, rabbit anti-glial fibrillary acidic protein (GFAP, 1∶1500 DAKO Cytomation, Glostrup Denmark), rabbit anti- Iba1 (1∶200, WAKO, Japan), and rat anti-mouse ED1 (CD68, 1∶1000, Nordic Biosite, Sweden), 1∶1000, overnight at 4°C overnight, and thereafter detected by incubation in secondary antibodies for 2 h. Secondary antibodies included were Texas Red-conjugated donkey anti-rabbit antibody (1∶200; Abcam, Cambridge, UK), Alexa 488 goat anti-rabbit IgG (Molecular Probes) and Alexa 564 goat anti-rat (Molecular Probes).

Techniques: Immunostaining, Control, Staining

Journal: PLoS ONE

Article Title: Silver and Gold Nanoparticles Exposure to In Vitro Cultured Retina – Studies on Nanoparticle Internalization, Apoptosis, Oxidative Stress, Glial- and Microglial Activity

doi: 10.1371/journal.pone.0105359

Figure Lengend Snippet: A–D. Iba1-immunohistochemistry revealing microglial cells with ramified (A), intermediate (B), round (C) and amoeboid (D) morphology, respectively. Quantification of the fractions of microglia in three different stages of activation was performed: intermediate or “early activated” cells as judged from the morphology and co-expression of ED1 while round- and amoeboid morphologies are regarded as “late activated” cells. The graph shows the distribution of activation stages, normalized to total numbers of Iba1-positive cells/explant; values ±SD (n = 4 explants/group) are given. * p <0.05 compared to control. GCL = ganglion cell layer; INL = inner nuclear layer; ONL = outer nuclear layer. Scale bars equal 50 µm.

Article Snippet: Sections were then incubated with primary antibodies, rabbit anti-glial fibrillary acidic protein (GFAP, 1∶1500 DAKO Cytomation, Glostrup Denmark), rabbit anti- Iba1 (1∶200, WAKO, Japan), and rat anti-mouse ED1 (CD68, 1∶1000, Nordic Biosite, Sweden), 1∶1000, overnight at 4°C overnight, and thereafter detected by incubation in secondary antibodies for 2 h. Secondary antibodies included were Texas Red-conjugated donkey anti-rabbit antibody (1∶200; Abcam, Cambridge, UK), Alexa 488 goat anti-rabbit IgG (Molecular Probes) and Alexa 564 goat anti-rat (Molecular Probes).

Techniques: Immunohistochemistry, Activation Assay, Expressing, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Neurodegeneration and Glial Response after Acute Striatal Stroke: Histological Basis for Neuroprotective Studies

doi: 10.1155/2016/3173564

Figure Lengend Snippet: Microglia activation revealed by anti-CD68 and MHC-II immunohistochemistry. Control animals injected with sterile saline (a-b, l-m). Morphological activation of CD68+ or MHC-II+ microglia at 3 (c-d, n-o), 7 (e-f, p-q), 14 (g-h, r-s), and 30 (i-j, t-u) PLDs. Both techniques are labeled activated round macrophages (arrows). Quantitative analysis showed maximum number of cells at 7 PLDs with decrease at later survival times (k, v). ∗ p < 0.05 compared to control; # p < 0.05 compared to previous survival time. Scale bars: (a), (c), (e), (g), (i), (l), (n), (p), (r), and (t) (100 μ m) and (b), (d), (f), (h), (j), (m), (o), (q), (s), and (u) (20 μ m).

Article Snippet: Activated microglia/macrophages were labeled using the antibody anti-rat CD68 (clone ED1, 1 : 500, Serotec, UK), which binds to an epitope on the lysosomal membrane of activated macrophages/microglia [ – ], rabbit anti-Iba1 (1 : 1000, WAKO), an antibody that recognizes a calcium binding protein present in the cytoplasm of microglia [ – ], and mouse anti-MHC-II (1 : 100, Serotec), an antibody that recognizes the major histocompatibility complex class II molecule [ ].

Techniques: Activation Assay, Immunohistochemistry, Control, Injection, Sterility, Saline, Labeling

Journal: Journal of Translational Medicine

Article Title: Dermal tissue remodeling and non-osmotic sodium storage in kidney patients

doi: 10.1186/s12967-019-1815-5

Figure Lengend Snippet: Correlations between clinical data, tissue remodeling responses, and dermal sodium

Article Snippet: Skin cryo sections were incubated for 1 h with the following primary antibodies/reagents: mouse anti-human α-smooth muscle actin (SMA; clone 1A4, Sigma-Aldrich, St Louis, USA), rabbit anti-human CD3 (clone A0452, Dako, Glostrup, Denmark), biotinylated hyaluronan binding protein (HABP, Seikagaku, Tokyo, Japan), mouse anti heparan sulphate mAB JM403 [ ], mouse anti-human podoplanin (Clone D240, ThermoFisher, Rockford, USA), mouse anti-human CD68 (clone ED1, AbD Serotec, Oxford, UK), rabbit anti-versican (ITK Diagnostics, B.V., Uithoorn, The Netherlands) and mouse anti-human MCP-1 (Peprotech, Rocky Hill, USA) diluted in PBS/1% Bovine Serum Albumin (BSA).

Techniques: Clinical Proteomics, Expressing, Concentration Assay