Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Characterization of immune cell subtypes in three commonly used mouse strains reveals gender and strain-specific variations

doi: 10.1038/s41374-018-0137-1

Figure Lengend Snippet: Antibody combinations used in immunophenotyping

Article Snippet: Fixation/Permeabilization Solution Kit with BD Golgi-PlugTM kit and Mouse Foxp3 Buffer Set was used for intracellular staining and purchased from BD Biosciences. table ft1 table-wrap mode="anchored" t5 caption a7 iMC CD11b Vio Blue Gr-1 APC-eFluor780 * Ly6b FITC Macrophages CD11b Vio Blue F4/80 APC CD68 PE mDC CD11b Vio Blue CD11c PE CD80 APC CD86 FITC pDC * CD11b Vio Blue CD11c PE B220 Vio770 Siglec H FITC CD4 T-cells CD3 APC-Cy7 CD4 eFluor450 CD4 T-regs CD3 APC-Cy7 CD4 eFluor450 CD25 PE FoxP3APC CD8 T-cells CD3 APC-Cy7 CD8 FITC B cells B220 Vio770 CD19 PE NK cells CD3 APC-Cy7 NKp46 FITC Neutrophils CD11b Vio Blue Gr-1 APC-eFluor780 Ly6b FITC * F4/80 APC Open in a separate window * negative tor this antibody Antibody combinations used in immunophenotyping table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Fluorochrome Vendor Clone Catalogue # Antibody Concentration (in 100 pL FACS buffer) CD3e APC-Cy7 Fisher Scientific 145–2C11 BDB557596 1 pL CD4 eFluor450 eBioscience GK1.5 48–0041-82 1 pL CD8 FITC eBioscience 53–6.7 11–0081-82 0.6 pL CD11b Vio Blue Miltenyi M1/70.15.11.5 130–097-336 3.75 pL CD11c PE Miltenyi N418 130102545 3.75 pL CD19 PE Miltenyi 6D5 130–102-598 5 pL CD25 PE eBioscience PC61.5 12–0251-81 0.5 pL CD68 PE Miltenyi FA-11 130–102-614 3.75 pL CD80 APC Miltenyi 16–10A1 130–102-584 3.75 pL CD86 FITC Miltenyi PO3.3 130–102-506 5 pL B220 APC-Vio770 Miltenyi RA3–6B2 130–102-267 3.75 pL F4/80 APC Miltenyi REA126 130–102-379 3.75 pL FoxP3 APC eBioscience FJK-16s 17–5773-82 5 pL Gr1 APC-eFluor780 eBioscience RB6–8C5 47–5931-80 1.2 pL Ly6b FITC abcam 7/4 ab53453 0.65 pL NKp46 FITC Miltenyi 29A1.4.9 130–102-300 3.75 pL SiglecH FITC eBioscience eBio440c 11–0333-82 0.5 pL Open in a separate window Details of antibodies used in the study

Techniques:

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Characterization of immune cell subtypes in three commonly used mouse strains reveals gender and strain-specific variations

doi: 10.1038/s41374-018-0137-1

Figure Lengend Snippet: Details of antibodies used in the study

Article Snippet: Fixation/Permeabilization Solution Kit with BD Golgi-PlugTM kit and Mouse Foxp3 Buffer Set was used for intracellular staining and purchased from BD Biosciences. table ft1 table-wrap mode="anchored" t5 caption a7 iMC CD11b Vio Blue Gr-1 APC-eFluor780 * Ly6b FITC Macrophages CD11b Vio Blue F4/80 APC CD68 PE mDC CD11b Vio Blue CD11c PE CD80 APC CD86 FITC pDC * CD11b Vio Blue CD11c PE B220 Vio770 Siglec H FITC CD4 T-cells CD3 APC-Cy7 CD4 eFluor450 CD4 T-regs CD3 APC-Cy7 CD4 eFluor450 CD25 PE FoxP3APC CD8 T-cells CD3 APC-Cy7 CD8 FITC B cells B220 Vio770 CD19 PE NK cells CD3 APC-Cy7 NKp46 FITC Neutrophils CD11b Vio Blue Gr-1 APC-eFluor780 Ly6b FITC * F4/80 APC Open in a separate window * negative tor this antibody Antibody combinations used in immunophenotyping table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Fluorochrome Vendor Clone Catalogue # Antibody Concentration (in 100 pL FACS buffer) CD3e APC-Cy7 Fisher Scientific 145–2C11 BDB557596 1 pL CD4 eFluor450 eBioscience GK1.5 48–0041-82 1 pL CD8 FITC eBioscience 53–6.7 11–0081-82 0.6 pL CD11b Vio Blue Miltenyi M1/70.15.11.5 130–097-336 3.75 pL CD11c PE Miltenyi N418 130102545 3.75 pL CD19 PE Miltenyi 6D5 130–102-598 5 pL CD25 PE eBioscience PC61.5 12–0251-81 0.5 pL CD68 PE Miltenyi FA-11 130–102-614 3.75 pL CD80 APC Miltenyi 16–10A1 130–102-584 3.75 pL CD86 FITC Miltenyi PO3.3 130–102-506 5 pL B220 APC-Vio770 Miltenyi RA3–6B2 130–102-267 3.75 pL F4/80 APC Miltenyi REA126 130–102-379 3.75 pL FoxP3 APC eBioscience FJK-16s 17–5773-82 5 pL Gr1 APC-eFluor780 eBioscience RB6–8C5 47–5931-80 1.2 pL Ly6b FITC abcam 7/4 ab53453 0.65 pL NKp46 FITC Miltenyi 29A1.4.9 130–102-300 3.75 pL SiglecH FITC eBioscience eBio440c 11–0333-82 0.5 pL Open in a separate window Details of antibodies used in the study

Techniques: Concentration Assay

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Characterization of immune cell subtypes in three commonly used mouse strains reveals gender and strain-specific variations

doi: 10.1038/s41374-018-0137-1

Figure Lengend Snippet: Cell suspensions from BM and spleen were isolated from 8-week old mice. The cells were stained with CD11b, F4/80 and CD68 antibodies for macrophages, CD11b and Gr-1 antibodies for iMCs, and CD11b and CD11c antibodies for mDCs. Ly6B was used as negative marker for iMCs. The cells were then subjected to flow cytometry to determine cell-type percentages. (A) iMCs in BM, (n≥7). (B) iMCs in the spleen, (n≥7). (C) Macrophages in BM, (n≥ 6). (D) Macrophages in spleen, (n≥9) (E) mDCs in BM, (n ≥ 10). (F) mDCs in the spleen, (n ≥ 13). Results are shown as scatter plots depicting average cell percentages (percent of live cells). Error bars denote SEM. Each dot represents the value from a single mouse. (♂) and (♀) represent male and female mice, respectively. **p ≤ 0.01, ***p ≤ 0.001, ****p≤0.0001.

Article Snippet: Fixation/Permeabilization Solution Kit with BD Golgi-PlugTM kit and Mouse Foxp3 Buffer Set was used for intracellular staining and purchased from BD Biosciences. table ft1 table-wrap mode="anchored" t5 caption a7 iMC CD11b Vio Blue Gr-1 APC-eFluor780 * Ly6b FITC Macrophages CD11b Vio Blue F4/80 APC CD68 PE mDC CD11b Vio Blue CD11c PE CD80 APC CD86 FITC pDC * CD11b Vio Blue CD11c PE B220 Vio770 Siglec H FITC CD4 T-cells CD3 APC-Cy7 CD4 eFluor450 CD4 T-regs CD3 APC-Cy7 CD4 eFluor450 CD25 PE FoxP3APC CD8 T-cells CD3 APC-Cy7 CD8 FITC B cells B220 Vio770 CD19 PE NK cells CD3 APC-Cy7 NKp46 FITC Neutrophils CD11b Vio Blue Gr-1 APC-eFluor780 Ly6b FITC * F4/80 APC Open in a separate window * negative tor this antibody Antibody combinations used in immunophenotyping table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Fluorochrome Vendor Clone Catalogue # Antibody Concentration (in 100 pL FACS buffer) CD3e APC-Cy7 Fisher Scientific 145–2C11 BDB557596 1 pL CD4 eFluor450 eBioscience GK1.5 48–0041-82 1 pL CD8 FITC eBioscience 53–6.7 11–0081-82 0.6 pL CD11b Vio Blue Miltenyi M1/70.15.11.5 130–097-336 3.75 pL CD11c PE Miltenyi N418 130102545 3.75 pL CD19 PE Miltenyi 6D5 130–102-598 5 pL CD25 PE eBioscience PC61.5 12–0251-81 0.5 pL CD68 PE Miltenyi FA-11 130–102-614 3.75 pL CD80 APC Miltenyi 16–10A1 130–102-584 3.75 pL CD86 FITC Miltenyi PO3.3 130–102-506 5 pL B220 APC-Vio770 Miltenyi RA3–6B2 130–102-267 3.75 pL F4/80 APC Miltenyi REA126 130–102-379 3.75 pL FoxP3 APC eBioscience FJK-16s 17–5773-82 5 pL Gr1 APC-eFluor780 eBioscience RB6–8C5 47–5931-80 1.2 pL Ly6b FITC abcam 7/4 ab53453 0.65 pL NKp46 FITC Miltenyi 29A1.4.9 130–102-300 3.75 pL SiglecH FITC eBioscience eBio440c 11–0333-82 0.5 pL Open in a separate window Details of antibodies used in the study

Techniques: Isolation, Staining, Marker, Flow Cytometry

Journal: Stem cells and development

Article Title: Neurogenic Niche Microglia Undergo Positional Remodeling and Progressive Activation Contributing to Age-Associated Reductions in Neurogenesis.

doi: 10.1089/scd.2015.0319

Figure Lengend Snippet: FIG. 3. Early age-related increased microglial activation in the V-SVZ, but not in non- neurogenic brain regions. (A) Quantification of proliferating BrdU-positive cells in the V- SVZ. n = 5 per age group. (B) Representative confocal im- ages of doublecortin-labeled V-SVZ wholemounts in 2- and 24-month-old mice. Scale bar: 50 mm. (C) Percentage of neuroblasts per field in wholemounts. n = 4 per group. (D) Confocal images from 2- to 12- and 24-month- old mice staining for Iba1 (green), CD68 (red), and DAPI (blue). Scale bar: 10mm. (E) Quantification of the number of Iba1+ cells in 2-, 6-, 12-, and 24-month-old mice in the V-SVZ. (F) Total area coveredbyIba1+cellsperfield assessed with ImageJ in the V- SVZ. (G) Size of CD68 reac- tivity in individual cells as- sessed with ImageJ. (H) CD68 reactivity in the cortex (Ctx) assessed with ImageJ. (I) CD68 reactivity in the stria- tum (Str) assessed with ImageJ n = 3 per group. Error bars= SEM. *P £ 0.05; xP £ 0.01; {P £ 0.001; ****P £ 0.0001.

Article Snippet: Before primary antibody incubation, the tissue was incubated in blocking solution containing 10% normal donkey serum in the appropriate PBST concentration overnight at 4 C. Incubation in primary antibodies was performed in blocking solution overnight at 4 C. The primary antibodies used were MASH1 (1:200; Cat. No. 556604; BD, Franklin Lakes, NJ), doublecortin (1:200; Cat. No. SCB066; Santa Cruz Biotechnology, Dallas, TX), Iba1 (1:200; Cat. No. 019-19741; Wako Chemicals, Richmond, VA), CD68 (1:250; Cat. No. MCA1957; AbD Serotec, Raleigh, NC), VCAM1 (1:50; Cat. No. 550547; BD), b-catenin (1:50; Cat. No. 610154; BD), CD31 (1:100; Cat. No. 550274; BD), bIII-Tubulin (1:500; Cat. No. MMS-435P-250; Covance, Austin, TX), and bromodeoxyuridine (BrdU; 1:250; Cat. No. NB500-169; Littleton, CO).

Techniques: Activation Assay, Labeling, Staining

Journal: Stem cells and development

Article Title: Neurogenic Niche Microglia Undergo Positional Remodeling and Progressive Activation Contributing to Age-Associated Reductions in Neurogenesis.

doi: 10.1089/scd.2015.0319

Figure Lengend Snippet: FIG. 6. Intraperitoneal inoculation with Bacillus Calmette–Gue´rin induces microglia activation and reduces neurogenesis in the V-SVZ. (A) Representative images of Iba1, EdU incorporation, and CD68 in V-SVZ wholemounts of mice 2 weeks after BCG inoculation or vehicle injected controls. Scale bar: 20 mm. (B) Quantification of CD68 immunoreactivity in the V-SVZ 2 weeks following inoculation with BCG. (C) Quantification of the number of EdU-positive cells in the V-SVZ in mice inoculated with BCG or vehicle. n = 3 per group. *P £ 0.05; xP £ 0.01. (D) Quantification of the different stages of microglia activation (as in Fig. 4A–D) in the V-SVZ wholemounts after 2 weeks of BCG inoculation (n = 3 per group). Error bars = SEM.

Article Snippet: Before primary antibody incubation, the tissue was incubated in blocking solution containing 10% normal donkey serum in the appropriate PBST concentration overnight at 4 C. Incubation in primary antibodies was performed in blocking solution overnight at 4 C. The primary antibodies used were MASH1 (1:200; Cat. No. 556604; BD, Franklin Lakes, NJ), doublecortin (1:200; Cat. No. SCB066; Santa Cruz Biotechnology, Dallas, TX), Iba1 (1:200; Cat. No. 019-19741; Wako Chemicals, Richmond, VA), CD68 (1:250; Cat. No. MCA1957; AbD Serotec, Raleigh, NC), VCAM1 (1:50; Cat. No. 550547; BD), b-catenin (1:50; Cat. No. 610154; BD), CD31 (1:100; Cat. No. 550274; BD), bIII-Tubulin (1:500; Cat. No. MMS-435P-250; Covance, Austin, TX), and bromodeoxyuridine (BrdU; 1:250; Cat. No. NB500-169; Littleton, CO).

Techniques: Activation Assay, Injection

Journal: Cell Death & Disease

Article Title: Large triglyceride-rich lipoproteins in hypertriglyceridemia are associated with the severity of acute pancreatitis in experimental mice

doi: 10.1038/s41419-019-1969-3

Figure Lengend Snippet: a Triglyceride concentration of plasma before caerulein treatment between wild-type, HTG1 (triglyceride concentration, 1000–2000 mg/dL) and HTG2 mice(triglyceride concentration, å 2000 mg/dL). b , c Plasma amylase and lipase activity at 12 th hour after the first injection of caerulein (50 μg/kg, 10 times, hourly) between these three groups. d Representative photomicrographs of H&E-stained section of pancreas in wild-type, HTG1 and HTG2 mice after acute pancreatitis induction. Scale bar = 500 or 100 μm. e Pathological scores of the pancreas in wild-type, HTG1 and HTG2 mice after acute pancreatitis induction. f Incidence rates of pancreatic necrosis between these three groups after acute pancreatitis induction. Each value was the mean ± SEM for n = 6. g Immunohistochemistry evaluation for myeloperoxidase and CD68 in pancreas between wild-type, HTG1 and HTG2 mice after acute pancreatitis induction. Scale bar = 100 μm. i Semiquantitative results of the area ratio of myeloperoxidase and CD68-positive cells among wild-type, HTG1 and HTG2 mice after acute pancreatitis induction. Each value was the mean ± SEM for n = 3–5. * p < 0.05 or ** p < 0.01 or *** p < 0.001 vs wild-type group. # p < 0.05 or ## p < 0.01 or ### p < 0.001 vs HTG1 group. WT wild-type, TG triglyceride, Ed edema, Necr necrosis, Vac vacuolisation, Infl inflammation, FFAs free fatty acids, MPO myeloperoxidase

Article Snippet: Neutrophil and macrophage infiltration were evaluated by immunohistochemical staining using anti-myeloperoxidase rabbit polyclonal antibody (Abcam, ab9535) and anti-CD68 rabbit polyclonal antibody (Boster, BA3638), respectively.

Techniques: Concentration Assay, Clinical Proteomics, Activity Assay, Injection, Staining, Immunohistochemistry

Journal: Frontiers in neurology

Article Title: Multiplex immunohistochemistry reveals cochlear macrophage heterogeneity and local auditory nerve inflammation in cisplatin-induced hearing loss.

doi: 10.3389/fneur.2022.1015014

Figure Lengend Snippet: FIGURE 2 Classification of mixed macrophages according to the intensity of expressed markers. (A) Watershed cell segmentation to identify the boundaries of single macrophages. Cochlea were obtained from 4-weeks old mice, and immunostained using the mIHC. The red line denotes the boundaries of each macrophage with F4/80 staining. (B) Quantification of the population in the resting state to identify the percentage of each category in the modiolus including auditory nerve and spiral ganglia. Mid-modiolar sections were analyzed, and macrophages population of each category, M0, M1-like, M2-like and M1/M2, were quantified (n = 5). The M0 non-activated macrophages represented the major percentage among all macrophages, with a significant difference compared to other categories. The mixed M1/M2 macrophages were presented minimally, while the M1-like and M2-like macrophages expressing single marker represented the least percentages. *P < 0.05, ****P < 0.0001, ns: not significant, by one-way ANOVA with Tukey’s multiple comparisons test. The error bars represent standard deviation from the mean. (C) Example of mixed macrophages in which M1 markers (CD68, CD86) expression was similar to that of the M2 markers (CD206, CD163). (D) A representative macrophage of a group of mixed macrophages in which the CD86 (M1 marker) showed the highest expression compared to the other markers. (E) Illustration of another group of mixed macrophages in which CD163 (M2 marker) had the highest expression among all markers. (F) Single-cell-based marker intensity for each mixed macrophage in the modiolus including auditory nerve and spiral ganglia. Each dot represents a single mixed macrophage (n = 74 from 7 cochleae); the mean intensity of M1 and M2 markers in each macrophage are plotted against each other. The cut-offvalue was set at 1, below which most of the macrophages were included, and the different types of macrophages were identified. The red box marks the area of the dominant cell population in which most of the macrophages represent M1 and M2 markers equally within the cut-offvalue. The numbers indicate the percentages of the cell population of each quadrant; in the lower right quadrant, M1-polarized cells expressing higher M1 marker intensities (>1) than M2 cells, 8.44, 3.90, 9.49, and 5.23%: the upper left quadrant, M2-polarized cells expressing higher M2 marker intensities (>1) than M1 cells, 6.56, 3.90, 10.83, and 9.78%: the upper right quadrant, cells strongly expressing both M1 and M2 markers, 2.56, 5.23, 1.51, and 3.90%.

Article Snippet: In brief, all primary antibodies in 0.1 PBS with 0.03% triton-X and 5% blocking solution (Nacalai Tesque) were incubated for 1 h at room temperature (23◦C) in the following sequence and concentration: F4/80 monocyte marker for the identification of Frontiers inNeurology 04 frontiersin.org cochlear macrophages (rat monoclonal, Abcam, Cat # ab6640, RRID # AB_1140040, 1:100) (11); CD68 lysosome-associated membrane protein marker (31) for the detection of inner earresident M1 macrophages (rabbit monoclonal, Cell Signaling Technology, Cat # 97778, 1:200) (19); CD206 mannose receptor C type 1 marker for the detection of M2 macrophages (goat polyclonal, R&D systems, Cat # AF2535, RRID # AB_2063012, 1:100) (32); CD86 marker for the detection of M1 macrophages (rabbit monoclonal, Cell Signaling Technology, Cat # 19589, RRID # AB_2892094, 1:100) (33); ionized calcium-binding adaptor molecule 1 (Iba1) microglia/macrophage marker (goat polyclonal, Abcam, Cat # ab5076, RRID # AB_2224402, 1:500) (9), CD163 marker for the detection of M2 macrophages (rabbit monoclonal EPR19518, Abcam, Cat # ab182422, RRID # AB_2753196, 1:200) (34), CXCR1 chemokine receptor (rabbit polyclonal, Bioss antibodies, Cat # bs-1009R, RRID # AB_10857682, 1:250) (35).

Techniques: Staining, Expressing, Marker, Standard Deviation

Journal: Frontiers in neurology

Article Title: Multiplex immunohistochemistry reveals cochlear macrophage heterogeneity and local auditory nerve inflammation in cisplatin-induced hearing loss.

doi: 10.3389/fneur.2022.1015014

Figure Lengend Snippet: FIGURE 4 Cisplatin increases the expression of M1, M2, and Iba1 markers in the auditory nerve. (A) Macrophage marker expression in different groups. On day 8 post-cisplatin injection, there was a marked increase in the expression of M1 markers (CD68 and CD86), M2 markers (CD206 and CD163), and Iba1. This increased expression was temporary and showed recovery on day 15. This increased expression suggested a temporary auditory nerve inflammation. There was no difference between the expressed markers in the control group. (B) Statistical analysis of the ratio of each marker expression in F4/80+ macrophages. In all the markers, there was significant increase in the ratio of expression on day 8 in cisplatin-injected mice (n = 10) compared to that of the day 8 control (n = 8), day 0 pre-injection (n = 6), and day 15 cisplatin-injected mice (n = 8). ****P < 0.0001, ns, not significant by two-way ANOVA with Bonferroni post hoc test. The error bars represent standard deviation from the mean.

Article Snippet: In brief, all primary antibodies in 0.1 PBS with 0.03% triton-X and 5% blocking solution (Nacalai Tesque) were incubated for 1 h at room temperature (23◦C) in the following sequence and concentration: F4/80 monocyte marker for the identification of Frontiers inNeurology 04 frontiersin.org cochlear macrophages (rat monoclonal, Abcam, Cat # ab6640, RRID # AB_1140040, 1:100) (11); CD68 lysosome-associated membrane protein marker (31) for the detection of inner earresident M1 macrophages (rabbit monoclonal, Cell Signaling Technology, Cat # 97778, 1:200) (19); CD206 mannose receptor C type 1 marker for the detection of M2 macrophages (goat polyclonal, R&D systems, Cat # AF2535, RRID # AB_2063012, 1:100) (32); CD86 marker for the detection of M1 macrophages (rabbit monoclonal, Cell Signaling Technology, Cat # 19589, RRID # AB_2892094, 1:100) (33); ionized calcium-binding adaptor molecule 1 (Iba1) microglia/macrophage marker (goat polyclonal, Abcam, Cat # ab5076, RRID # AB_2224402, 1:500) (9), CD163 marker for the detection of M2 macrophages (rabbit monoclonal EPR19518, Abcam, Cat # ab182422, RRID # AB_2753196, 1:200) (34), CXCR1 chemokine receptor (rabbit polyclonal, Bioss antibodies, Cat # bs-1009R, RRID # AB_10857682, 1:250) (35).

Techniques: Expressing, Marker, Injection, Control, Standard Deviation

Journal: Aging (Albany NY)

Article Title: Scorpion and centipede alleviates severe asthma through M2 macrophage-derived exosomal miR-30b-5p

doi: 10.18632/aging.204053

Figure Lengend Snippet: SC improves severe asthma. ( A ) H&E staining of lung tissues in the SA mouse model treated with SC. Scale bars = 100 μm. ( B ) The expression level of TNF-α in lung tissues was detected by ELISA assay. ( n = 5) ( C ) The expression level of IL-1β in lung tissues was detected by ELISA assay. ( n = 5) ( D ) The shape and size of the particles were evaluated using transmission electron microscopy. Representative pictures were shown. Scale bars = 100 nm. ( E ) Diameter distribution and particle concentration of exosomes were observed by nanoanalysis. ( F ) The expression levels of exosome markers and macrophage markers were detected by Western blotting. ( G ) Flow cytometry analysis of CD163+/CD68+ ratio. * P < 0.05; ** P < 0.01.

Article Snippet: Next, the cells were blocked with 1% bovine serum albumin (GIBCO, cat#23208) and were incubated with primary antibodies against CD68 (1:1000, Santa Cruz, cat#sc-52998) and Arg-1 (1:500; Cell Signaling Technology, 93668S) at 4°C overnight.

Techniques: Staining, Expressing, Enzyme-linked Immunosorbent Assay, Transmission Assay, Electron Microscopy, Concentration Assay, Western Blot, Flow Cytometry

Journal: Antioxidants

Article Title: Anthocyanins Protect Hepatocytes against CCl 4 -Induced Acute Liver Injury in Rats by Inhibiting Pro-inflammatory mediators, Polyamine Catabolism, Lipocalin-2, and Excessive Proliferation of Kupffer Cells

doi: 10.3390/antiox8100451

Figure Lengend Snippet: The impact of CCl 4 and the bilberry fruit extract on pro-inflammatory mediators in liver homogenate a .

Article Snippet: The liver homogenate concentration level of Lipocalin-2 (Neutrophil gelatinase-associated lipocalin), CD68 (Cluster of Differentiation 68), TNF-α (tumor necrosis factor-alpha), IL-4 (interleukin-4), IL-6 (interleukin-6), and IL-13 (interleukin-13) were determined using a commercial ELISA kit (Rat NGAL ELISA kit, ab207925, Abcam, USA; Rat CD68 ELISA kit, CSB-E13297r, Cusabio, USA; Rat TNF-α ELISA kit, Antibodies-online, Germany; Rat IL-4 ELISA kit, Antibodies-online, Germany; Rat IL-6 ELISA kit, R&D Systems, USA; Rat IL-13 ELISA kit, Antibodies-online, Germany) according to the manufacturer’s instructions.

Techniques:

Journal: Antioxidants

Article Title: Anthocyanins Protect Hepatocytes against CCl 4 -Induced Acute Liver Injury in Rats by Inhibiting Pro-inflammatory mediators, Polyamine Catabolism, Lipocalin-2, and Excessive Proliferation of Kupffer Cells

doi: 10.3390/antiox8100451

Figure Lengend Snippet: Effect of CCl 4 and bilberry fruit extract on histopathology of rat liver after 24 h (H&E and Immunohistochemical staining). I -group I (untreated group), II -group II (treated group), III -group III (CCl 4 ), IV -group IV (treated + CCl 4 ). ( A ) group I (untreated group), 100×; ( B ) group II (treated group), 100×; ( C1 – C11 ) group III (CCl 4 ); C1-Necrosis followed by minor and severe reversible change, 100×; C2-Necrosis followed by macrovesicular hepatocytes, 100×; C3-Necrosis followed by vacuolar hepatocytes, 100×; C4-Hemorrhagic coagulation necrosis, 100×; C5-Necrosis (🞲), macrovesicular hepatocytes (→), hydropic hepatocytes ( → ), inflammatory mononuclear infiltrate ( 🞲 ), 200×; C6-Hemorrhagic coagulation necrosis (🞲), 200×; C7-Necrosis (🞲), vacuolar hepatocytes (→), 200×; C8- macrovesicular fatty hepatocytes (→), 400×; C9-Necrosis (🞲), microvesicular fatty hepatocytes (→), 400×; C10-Necrosis (🞲), 400×; C11-Inflammatory mononuclear infiltrate (🞲), 400×; ( D1 – D3 ) group IV (treated + CCl 4 ); D1-minor reversible change (hydropic hepatocytes), 100×; D2- minor reversible change (vacuolar hepatocytes) and extended sinusoidal spaces, 200×; D3-Hydropic hepatocytes (→), 400×; ( E ) immunohistochemical detection of COX-2, 100×; ( F ) immunohistochemical detection of iNOS (NOS2), 100×; ( G ) immunohistochemical detection of TNF-α, 100×, ( H ) immunohistochemical detection of NGAL, 100×; ( I ) immunohistochemical detection Kupffer cells identification marker-CD68, 100×; ( J ) morphometric analysis extent of the necrotic area on a selected H&E field in the liver. The data show the average value ± S.D. (%) for 10 fields at the magnification of 200× for each animal in each group (H&E staining); ( K ) semiquantitative evaluation of COX-2, iNOS, TNF-α, NGAL, and CD68 immunohistochemistry staining. Staining intensity was graded as: - (negative), +/- (weak positive), + (positive), ++ (strongly positive), or +++ (very strongly positive).

Article Snippet: The liver homogenate concentration level of Lipocalin-2 (Neutrophil gelatinase-associated lipocalin), CD68 (Cluster of Differentiation 68), TNF-α (tumor necrosis factor-alpha), IL-4 (interleukin-4), IL-6 (interleukin-6), and IL-13 (interleukin-13) were determined using a commercial ELISA kit (Rat NGAL ELISA kit, ab207925, Abcam, USA; Rat CD68 ELISA kit, CSB-E13297r, Cusabio, USA; Rat TNF-α ELISA kit, Antibodies-online, Germany; Rat IL-4 ELISA kit, Antibodies-online, Germany; Rat IL-6 ELISA kit, R&D Systems, USA; Rat IL-13 ELISA kit, Antibodies-online, Germany) according to the manufacturer’s instructions.

Techniques: Histopathology, Immunohistochemical staining, Staining, Coagulation, Marker, Immunohistochemistry

Journal: Diabetology & Metabolic Syndrome

Article Title: Liraglutide attenuates atherosclerosis via inhibiting ER-induced macrophage derived microvesicles production in T2DM rats

doi: 10.1186/s13098-017-0289-y

Figure Lengend Snippet: Expression of CD68, TUNEL and caspase-3 and microvesicles concentration in different groups. a Immunohistochemistry staining for CD68 and TUNEL. Scale bar: 50 μm. b Western blot for caspase-3. c Microvesicles concentration. Values are mean ± SD. n = 8, n = 11 and n = 11 for control group, diabetes group and liraglutide group respectively ( b , c ). * P < 0.05, ** P < 0.01 compared with control group ( b , c ). # P < 0.05 compared with diabetes group ( b , c )

Article Snippet: Sections were stained for CD68 with a rat anti-CD68 monoclonal antibody (at 1:200 dilution, sc-101447, Santa Cruz, America) and the avidin–biotin–peroxidase complex technique.

Techniques: Expressing, TUNEL Assay, Concentration Assay, Immunohistochemistry, Staining, Western Blot, Control