Journal: Journal of Exercise Rehabilitation

Article Title: Effect of combined intervention of exercise and autologous bone marrow stromal cell transplantation on neurotrophic factors and pain-related cascades over time after sciatic nerve injury

doi: 10.12965/jer.2244006.003

Figure Lengend Snippet: At early stage of axonal regeneration, combined intervention of treadmill exercise and autologous bone marrow stromal cell (BMSC) transplantation regulates induction levels of nuclear factor (NF)-κB, interleukin (IL)-6, and tumor necrosis factor (TNF)-α in the ipsilateral lumbar 4 and lumbar 5 dorsal root ganglion. (A) Western blot results on TNF-α, NF-κB, and IL-6 at 1 and 2 weeks after sciatic nerve injury. (B) Quantitative graph on TNF-α/actin ratio at week 1 and 2 later. (C) Quantitative graph on NF-κB/actin ratio at week 1 and 2 later. (D) Quantitative graph on IL-6/actin ratio at week 1 and 2 later. CONT, normal control; SS, sedentary group; SE, low-intensity treadmill exercise group; SB, BMSC transplantation group; SBE, BMSC transplantation group+treadmill exercise. * P <0.05, ** P <0.01, *** P <0.001 vs. CONT group. # P <0.05, ## P <0.01, ### P <0.001 vs. SS group. †† P <0.01 vs. SE group.

Article Snippet: Protein (20 μg) was used for Western blot analysis using anti-TrkB rabbit polyclonal antibody (1:1,000, Cell Signaling Biotechnology, Danvers, MA, USA), anti-β-actin mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-NF-κB mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-CNTF mouse monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-NGF mouse monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-BDNF mouse monoclonal antibodies (1:1,000, Santa Cruz Biotechnology), anti-TNF-α rabbit polyclonal antibody (1:1,000, Sino Biological, Wayne, PA, USA), anti-IL-6 rabbit polyclonal antibody (1:1,000, GeneTex Inc., Irvine, CA, USA).

Techniques: Transplantation Assay, Western Blot

Journal: Journal of Exercise Rehabilitation

Article Title: Effect of combined intervention of exercise and autologous bone marrow stromal cell transplantation on neurotrophic factors and pain-related cascades over time after sciatic nerve injury

doi: 10.12965/jer.2244006.003

Figure Lengend Snippet: At late stage of axonal regeneration, combined intervention of treadmill exercise and autologous bone marrow stromal cell (BMSC) transplantation regulates activation of proinflammatory cytokines in the ipsilateral lumbar 4 and lumbar 5 dorsal root ganglion. (A) Western blot results on tumor necrosis factor (TNF)-α, nuclear factor (NF)-κB, and interleukin (IL)-6 at 3 and 5 weeks after sciatic nerve injury. (B) Quantitative graph on TNF-α/actin ratio at week 3 and 5 later. (C) Quantitative graph on NF-κB/actin ratio at week 3 and 5 later. (D) Quantitative graph on IL-6/actin ratio at week 3 and 5 later. CONT, normal control; SS, sedentary group; SE, low-intensity treadmill exercise group; SB, BMSC transplantation group; SBE, BMSC transplantation group+treadmill exercise. * P <0.05, *** P <0.001 vs. CONT group. # P <0.05, ## P <0.01, ### P <0.001 vs. SS group.

Article Snippet: Protein (20 μg) was used for Western blot analysis using anti-TrkB rabbit polyclonal antibody (1:1,000, Cell Signaling Biotechnology, Danvers, MA, USA), anti-β-actin mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-NF-κB mouse monoclonal antibody (1:1,000, Santa Cruz Biotechnology), anti-CNTF mouse monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-NGF mouse monoclonal antibody (1:1,000, Cell Signaling Biotechnology), anti-BDNF mouse monoclonal antibodies (1:1,000, Santa Cruz Biotechnology), anti-TNF-α rabbit polyclonal antibody (1:1,000, Sino Biological, Wayne, PA, USA), anti-IL-6 rabbit polyclonal antibody (1:1,000, GeneTex Inc., Irvine, CA, USA).

Techniques: Transplantation Assay, Activation Assay, Western Blot

Journal: Neuropsychiatric Disease and Treatment

Article Title: Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk

doi: 10.2147/NDT.S154809

Figure Lengend Snippet: Flow diagram of the study. Abbreviations: Erk, extracellular signal-regulated kinase; PCR, polymerase chain reaction; SAH, subarachnoid hemorrhage; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Subsequently, the blots were separately incubated overnight at 4°C with the following antibodies: rabbit anti-caspase-3 (1:1,000; Cell Signaling Technology, #9665, Danvers, MA, USA), rabbit anti-bax (1:1,000; Cell Signaling Technology, #2772), rabbit anti-bcl-2 (1:1,000; Cell Signaling Technology, #2870), rabbit anti-phosphorylated Erk (p-Erk, 1:2,000; Cell Signaling Technology, #4370), rabbit anti-Erk (1:2,000; Cell Signaling Technology, #4695), rabbit anti-TNF-α (1:1,000; R&D Systems, AF-510-SP, Minneapolis, MN, USA) or rabbit anti-β-actin (1:1,000, as a control; Cell Signaling Technology, #4970).

Techniques: Polymerase Chain Reaction

Journal: Neuropsychiatric Disease and Treatment

Article Title: Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk

doi: 10.2147/NDT.S154809

Figure Lengend Snippet: The effects of TNF-α antibody on the expression of caspase-3, bax and bcl-2 in the hypothalamus after SAH. Notes: Rats were microinfused with anti-TNF-α antibody, or the same volume of PBS, 30 min before surgery. The hypothalamus was dissected out 48 h later. ( A ) Representative immunoblots and quantitative analyses showing the protein levels of TNF-α (n=7/group) relative to β-actin in the hypothalamus. ( B ) Changes in caspase-3 mRNA (n=6–7/group), bax mRNA (n=5–7/group) and bcl-2 mRNA (n=5/group). Representative immunoblots and quantitative analyses showing the protein levels of ( C ) caspase-3 (n=7/group), ( D ) bax (n=8–9/group) and ( E ) bcl-2 (n=6–10/group) relative to β-actin in the hypothalamus (* p <0.05, ** p <0.01 versus sham-operated control group, # p <0.05 versus PBS group). Abbreviations: SAH, subarachnoid hemorrhage; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Subsequently, the blots were separately incubated overnight at 4°C with the following antibodies: rabbit anti-caspase-3 (1:1,000; Cell Signaling Technology, #9665, Danvers, MA, USA), rabbit anti-bax (1:1,000; Cell Signaling Technology, #2772), rabbit anti-bcl-2 (1:1,000; Cell Signaling Technology, #2870), rabbit anti-phosphorylated Erk (p-Erk, 1:2,000; Cell Signaling Technology, #4370), rabbit anti-Erk (1:2,000; Cell Signaling Technology, #4695), rabbit anti-TNF-α (1:1,000; R&D Systems, AF-510-SP, Minneapolis, MN, USA) or rabbit anti-β-actin (1:1,000, as a control; Cell Signaling Technology, #4970).

Techniques: Expressing, Western Blot

Journal: Neuropsychiatric Disease and Treatment

Article Title: Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk

doi: 10.2147/NDT.S154809

Figure Lengend Snippet: Immunohistochemistry showing the changes of caspase-3, bax and bcl-2 in the hypothalamus 48 h after SAH. Notes: Rats were treated as described in . Representative coronal sections are also shown. Scale bar =100 μm. Abbreviations: SAH, subarachnoid hemorrhage; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Subsequently, the blots were separately incubated overnight at 4°C with the following antibodies: rabbit anti-caspase-3 (1:1,000; Cell Signaling Technology, #9665, Danvers, MA, USA), rabbit anti-bax (1:1,000; Cell Signaling Technology, #2772), rabbit anti-bcl-2 (1:1,000; Cell Signaling Technology, #2870), rabbit anti-phosphorylated Erk (p-Erk, 1:2,000; Cell Signaling Technology, #4370), rabbit anti-Erk (1:2,000; Cell Signaling Technology, #4695), rabbit anti-TNF-α (1:1,000; R&D Systems, AF-510-SP, Minneapolis, MN, USA) or rabbit anti-β-actin (1:1,000, as a control; Cell Signaling Technology, #4970).

Techniques: Immunohistochemistry

Journal: Neuropsychiatric Disease and Treatment

Article Title: Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk

doi: 10.2147/NDT.S154809

Figure Lengend Snippet: The effects of anti-TNF-α antibody on the levels of p-Erk in the hypothalamus 48 h after SAH. Notes: ( A ) The levels of p-Erk, Erk and bax in the control group, vehicle group and U0126 group were detected by Western blotting. ( B ) Quantitative analyses showing the levels of p-Erk relative to total Erk, and bax relative to β-actin normalized to the sham-operated control group (n=5/group, * p <0.05 versus sham-operated control group). ( C ) Levels of p-Erk, Erk and bax in the control group, vehicle group, anti-TNF-α antibody group and anti-TNF-α antibody+PMA group were detected by Western blotting. ( D ) Quantitative analyses showing the levels of p-Erk relative to total Erk, and bax relative to β-actin normalized to sham-operated control group (n=5/group, ** p <0.01 versus sham-operated control group). Abbreviations: Erk, extracellular signal-regulated kinase; PMA, phorbol-12-myristate-13-acetate; SAH, subarachnoid hemorrhage; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Subsequently, the blots were separately incubated overnight at 4°C with the following antibodies: rabbit anti-caspase-3 (1:1,000; Cell Signaling Technology, #9665, Danvers, MA, USA), rabbit anti-bax (1:1,000; Cell Signaling Technology, #2772), rabbit anti-bcl-2 (1:1,000; Cell Signaling Technology, #2870), rabbit anti-phosphorylated Erk (p-Erk, 1:2,000; Cell Signaling Technology, #4370), rabbit anti-Erk (1:2,000; Cell Signaling Technology, #4695), rabbit anti-TNF-α (1:1,000; R&D Systems, AF-510-SP, Minneapolis, MN, USA) or rabbit anti-β-actin (1:1,000, as a control; Cell Signaling Technology, #4970).

Techniques: Western Blot

Journal: Neuropsychiatric Disease and Treatment

Article Title: Anti-TNF-alpha antibody attenuates subarachnoid hemorrhage-induced apoptosis in the hypothalamus by inhibiting the activation of Erk

doi: 10.2147/NDT.S154809

Figure Lengend Snippet: The effects of anti-TNF-α antibody on anxiety-like behavior. Notes: Rats were treated as described in . Open field analysis was performed on both days 2 and 7 after SAH. ( A ) Time spent in the center and ( B ) the total distance traveled were respectively recorded (n=8/group, * p <0.05, ** p <0.01 versus sham-operated control group; # p <0.05, ## p <0.01 versus PBS group). Abbreviations: SAH, subarachnoid hemorrhage; TNF-α, tumor necrosis factor-alpha.

Article Snippet: Subsequently, the blots were separately incubated overnight at 4°C with the following antibodies: rabbit anti-caspase-3 (1:1,000; Cell Signaling Technology, #9665, Danvers, MA, USA), rabbit anti-bax (1:1,000; Cell Signaling Technology, #2772), rabbit anti-bcl-2 (1:1,000; Cell Signaling Technology, #2870), rabbit anti-phosphorylated Erk (p-Erk, 1:2,000; Cell Signaling Technology, #4370), rabbit anti-Erk (1:2,000; Cell Signaling Technology, #4695), rabbit anti-TNF-α (1:1,000; R&D Systems, AF-510-SP, Minneapolis, MN, USA) or rabbit anti-β-actin (1:1,000, as a control; Cell Signaling Technology, #4970).

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Suppression of MyD88-dependent signaling alleviates neuropathic pain induced by peripheral nerve injury in the rat

doi: 10.1186/s12974-017-0822-9

Figure Lengend Snippet: List of primary and secondary antibodies used for Western blot analysis

Article Snippet: TNF-α , rabbit , Sino Biological , 80045-RP02 , 1:500 , Overnight 4 °C.

Techniques: Western Blot, Incubation

Journal: Journal of Neuroinflammation

Article Title: Suppression of MyD88-dependent signaling alleviates neuropathic pain induced by peripheral nerve injury in the rat

doi: 10.1186/s12974-017-0822-9

Figure Lengend Snippet: Suppressed activation of glial cells and TNF-α production by inhibition of MyD88 in rat SDH after CCI. a Immunostaining showing inhibitory effects of MIP on activation of microglial cells (IBA1), and astrocytes (GFAP). Scale bar: 20 μm. b Western blot showing inhibitory effects of MIP on CCI-induced increased protein level of IBA1, GFAP, and TNF-α. c Data summary of B. Others are the same as Fig.

Article Snippet: TNF-α , rabbit , Sino Biological , 80045-RP02 , 1:500 , Overnight 4 °C.

Techniques: Activation Assay, Inhibition, Immunostaining, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Suppression of MyD88-dependent signaling alleviates neuropathic pain induced by peripheral nerve injury in the rat

doi: 10.1186/s12974-017-0822-9

Figure Lengend Snippet: Schematic illustration demonstrates MyD88-dependent signaling pathways of neuropathic pain induced by CCI. Nerve injury produces abundant HMGB1and IL-1β in the DRG and SDH. The binding of HMGB1 and IL-1β to their receptors (TLR2/4 andIL-1R, respectively) activates MyD88 in the DRG and SDH, which phosphorylate NF-κB p65 and ERK. Phosphorylated NF-κB p65 subsequently enter the nucleus to regulate the expression of proinflammation cytokines such as TNF-α. Phosphorylated ERK enter the nucleus to induce transcription factors such as AP-1, which regulates the expression of certain cytokines and activates glial cells. All these signaling events consequently result in central and peripheral sensitizations that produce neuropathic pain

Article Snippet: TNF-α , rabbit , Sino Biological , 80045-RP02 , 1:500 , Overnight 4 °C.

Techniques: Binding Assay, Expressing

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure1. Microglial–macrophageproductionofTNFafterfocalcerebralischemia.A,Immunofluorescencephotomicrographs of CD11b cells (red) in nonischemic cortex, in the peri-infarct region, and within infarct in a C57BL/6 mouse 24 h after pMCAo, with DAPI (blue) as a nuclear counterstain. B, Levels of TNF mRNA, measured by quantitative PCR and normalized to HPRT1, increased over time in ischemic hemispheres of C57BL/6 mice relative to control (Ctl) mice and sham-operated mice, reaching statisticallysignificantvalues6hafterpMCAo.ResultsareexpressedasmeanSD(n6–11).**p0.01;***p0.001.C, Combined in situ hybridization for TNF mRNA (red) and immunofluorescence for NeuN (green) in a C57BL/6 mouse 6 h after pMCAo, with DAPI as a nuclear counterstain, demonstrating that TNF mRNA (indicated by an arrow) is not expressed by NeuN

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Real-time Polymerase Chain Reaction, Control, In Situ Hybridization, Immunofluorescence

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure 3. TNF is expressed in microglia and infiltrating leukocytes. A, Immunofluorescence photomicrographs of the peri- infarct area in a chimeric mouse with infiltrating BM-derived GFP cells (green) stained for CD11b (red) 24 h after pMCAo. BM-derived,GFP cellsareprimarilylocatedwithininfarctandinperi-infarctregions,andallGFP cellscolocalizewithCD11b, withthecombinedgreenandredfluorescenceshownasyellow(arrows).B,Immunofluorescencephotomicrographsofachimeric mouse with infiltrating, BM-derived GFP cells (green) stained for TNF (red) 24 h after pMCAo. TNF is expressed primarily by residentmicroglia,becausemostredandgreencellsdonotcoincide,butalsobyinfiltratingGFP cells,becauseGFP cellsalso stain for TNF (arrow). C, Flow cytometry profiles gated on CD11b CD45 cells show increased infiltration of GFP cells in ischemic cortex of a BM-chimeric mouse 24 h after pMCAo, compared with limited numbers of GFP cells in cortex of an unmanipulated mouse. Note that the majority of infiltrating cells in ischemic cortex are GFP CD11b CD45 high cells. The bar graph shows the number of GFP CD11b CD45 dim and GFP CD11b CD45 high cells and reveals a significant recruitment of BM-derivedGFP cells24hafterpMCAo(one-wayANOVAfollowedbyDunnett’smultiplecomparisontest;n3–5pergroup). D,FlowcytometryprofilesshowingTNF CD11b CD45 dimmicrogliaandTNF CD11b CD45 highleukocytesinischemiccortex 24hafterpMCAoinaC57BL/6mouse.Quantificationrevealsalesion-specificincreaseinthetotalnumberofTNF cellsovertime (one-way ANOVA followed by Dunnett’s multiple comparison test; n 3–10). E, Quantification of the total number of CD11b CD45 dimmicrogliainunmanipulatedC57BL/6andGFP-BM-chimericmiceshowedthatchimericmicehavesignificantly fewer microglia in unmanipulated cortex compared with C57BL/6 mice, whereas the total number of leukocytes was unaffected by irradiation (one-way ANOVA followed by Tukey’s test for multiple comparisons; n 4–7). F, Flow cytometric evaluation showsthattheproportionofTNF CD11b CD45 cellswassimilarinGFP-BM-chimericandC57BL/6mice(n4–7).Results are expressed as mean SD. *p 0.05; **p 0.01; ***p 0.001. Scale bars, 20 m. Ctl, Control.

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Derivative Assay, Staining, Flow Cytometry, Comparison, Irradiation, Control

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure 4. TNF is neuroprotective after permanent focal cerebral ischemia. A, Toluidine blue staining of brain sections from TNF-KO,B6129,andC57BL/6mice24hafterpMCAo.IF,Infarct.B,EstimationofcorticalinfarctvolumeshowedthatTNF-KOmice developed significantly larger infarcts compared with WT B6129 and WT C57BL/6 mice (Mann–Whitney test; n 14–15). C, ToluidinebluestainingofbrainsectionsfromTNF-KOandC57BL/6mice5dafterpMCAo.D,TNF-KOmicedevelopedsignificantly larger infarcts compared with WT C57BL/6 mice (Mann–Whitney test; n 9–12) also at 5 d. Results are expressed as mean SD. *p 0.05. Scale bar, 1 mm.

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Staining, MANN-WHITNEY

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure5. Microglial-derivedTNFisneuroprotectivethroughtheTNF-p55receptor.A,ToluidinebluestainingofsectionsfromBM-chimericmice24hafterpMCAo.B,Estimationofcorticalinfarct volumeinBM-chimericmiceshowsthatKO-to-WTmicedevelopedinjuriessimilarlysizedtothoseofWT-to-WTmice( p0.26,Mann–Whitneytest;n15–20),showingthatTNFproducedby macrophageshadnoinfluenceontissueinjuryafterischemia.Furthermore,WT-to-KOmicedevelopedinjuriessimilarlysizedtothoseofKO-to-KOmice( p0.38,Mann–Whitneytest;n18), showing that macrophage-derived TNF could not compensate for the deficiency of microglial-derived TNF in KO mice and demonstrating a neuroprotective role of microglial-derived TNF. C, TNF staining of sections from ischemic BM-chimeric mice 24 h after pMCAo. TNF protein was present in all BM-chimeric mice, except in KO-to-KO BM-chimericmice. D, Relative TNF mRNA levels in the ipsilateral,ischemichemisphere24hafterpMCAofromKO-to-WT,WT-to-WT,KO-to-KO,andWT-to-KOmice.TNFmRNAwaspresentinKO-to-WTmice,WT-to-WTmice,and,toalesserextent,in WT-to-KO mice, but not in KO-to-KO mice. Normalized mRNA levels were calibrated relative to a pool of cDNA from unmanipulated C57BL/6 control mice (n 4–9). E, Toluidine blue staining of sections from TNF-p55R-KO (R1-KO), TNF-p75R-KO (R2-KO), TNF-p55p75R-KO (R-KO), and C57x129 mice 5 d after pMCAo. F, R1-KO mice developed significantly larger infarcts compared to both R2-KOandC57x129micebutnottoR-KOmice5dafterpMCAo(one-wayANOVAfollowedbyDunn’smultiplecomparisontest;n6–7).ResultsareexpressedasmeanSD.*p0.05;**p 0.01; ***p 0.001. Scale bars: A, E, 1 mm; C, 20 m. IF, Infarct; ND, none detected.

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Derivative Assay, Staining, Control

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure 6. Cellular recruitment and TLR2 expression in TNF-KO mice. A, Flow cytometry profiles gated on FSC/SSC show an increased recruitment of CD11b CD45 high leukocytes to the ischemic cortex from C57BL/6 and TNF-KO mice, 24 h after MCA occlusionrelativetounmanipulatedmice(Ctl;toprightquadrants).LeukocyteswereclearlydistinguishedfromCD11b CD45 dim

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Expressing, Flow Cytometry

Journal: Journal of Neuroscience

Article Title: Microglia Protect Neurons against Ischemia by Synthesis of Tumor Necrosis Factor

doi: 10.1523/jneurosci.5505-08.2009

Figure Lengend Snippet: Figure7. Activationofcaspase-3afterpermanentfocalcerebralischemia.Immunostaining for activated caspase-3 (Cas-3) cells shows that Cas-3 was expressed throughout the infarct at 24 h in cells with a microglial morphology (inset) in both TNF-KO and C57BL/6 mice. Immuno- fluorescence photomicrographs of the peri-infarct area in a C57BL/6 mouse shows colocaliza- tionbetweenCas-3(red)andCD11b cells24hafterpMCAo.AllCas-3 cellscolocalizedwith CD11b, with the combined green and red fluorescence shown as yellow (arrow). Immunofluo- rescencephotomicrographsoftheinfarctinaC57BL/6mousestainedforCas-3(red,arrow)and NeuN (green) 24 h after MCA occlusion are shown. Cas-3 is not expressed by neurons, because red fluorescence and green fluorescence do not coincide. Scale bars: 20 m; insert, 10 m.

Article Snippet: Antibodies used for staining were rat anti-mouse TNF (Endogen), rabbit anti-mouse TNF (Endogen), rat anti-mouse CD11b (Serotec), biotinylated mouse anti-NeuN (Millipore Bioscience Research Reagents), rat anti-mouse Gr-1 (Ly-6G and Ly-6C; Serotec), rabbit anti-caspase 3 (Cell Signaling Technology), and rabbit anti-p-NF B p65 (Ser536) (Santa Cruz Biotechnology).

Techniques: Immunostaining, Fluorescence