neun-immunoreactive structures Search Results


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  • 99
    Thermo Fisher dapi
    Dapi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 155801 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore α syn
    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in <t>MBP-α-syn</t> tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia
    α Syn, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 705 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cybermetrics optimas 6 5 software
    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in <t>MBP-α-syn</t> tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia
    Optimas 6 5 Software, supplied by Cybermetrics, used in various techniques. Bioz Stars score: 89/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    NEN Life Science tyramide signal amplification direct red system
    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in <t>MBP-α-syn</t> tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia
    Tyramide Signal Amplification Direct Red System, supplied by NEN Life Science, used in various techniques. Bioz Stars score: 91/100, based on 739 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Carl Zeiss axiom1 light microscope
    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in <t>MBP-α-syn</t> tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia
    Axiom1 Light Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 89/100, based on 281 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti neun
    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in <t>MBP-α-syn</t> tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia
    Anti Neun, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore glial fibrillary acidic protein
    Delivery of the 3RT-apoB antibody reduces neurodegeneration in the 3RTau tg model. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. The neocortex and hippocampus were immunostained with antibodies against the (a) astrocyte glial fibrillary acidic protein <t>(GFAP),</t> (c) neuronal marker NeuN, (e) and dendritic marker <t>MAP2</t> (green). (b) GFAP immunoreactivity expressed as corrected optical density of sections. (d) Stereological estimates (dissector method) of total NeuN-positive neuronal counts in the frontal cortex. (f) Image analysis of % area of the neuropil covered by MAP2 post-synaptic processes. * indicates statistical significance p
    Glial Fibrillary Acidic Protein, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3353 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Olympus 20x magnification
    Delivery of the 3RT-apoB antibody reduces neurodegeneration in the 3RTau tg model. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. The neocortex and hippocampus were immunostained with antibodies against the (a) astrocyte glial fibrillary acidic protein <t>(GFAP),</t> (c) neuronal marker NeuN, (e) and dendritic marker <t>MAP2</t> (green). (b) GFAP immunoreactivity expressed as corrected optical density of sections. (d) Stereological estimates (dissector method) of total NeuN-positive neuronal counts in the frontal cortex. (f) Image analysis of % area of the neuropil covered by MAP2 post-synaptic processes. * indicates statistical significance p
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    Elan Drug Technologies 3d6
    PGRN overexpression in hippocampus decreases amyloid plaque load in 5xFAD mice. ( a ) Representative images of the contralateral and ipsilateral hemispheres of Lenti-Ctrl- and Lenti-PGRN-injected 5xFAD mice. Dashed lines indicate boundaries used for quantification of dentate gyrus (DG) and non-dentate gyrus hippocampal (non-DG HP) areas. PGRN (red), <t>3D6</t> + plaques (green), and ThioS + plaques (magenta). Scale bar, 200 μm. ( b ) Quantification of PGRN immunostaining normalized to the uninjected contralateral hemisphere for Lenti-Ctrl- or Lenti-PGRN-injected mice. Two Lenti-PGRN-injected mice lacking sufficient overexpression (i.e.
    3d6, supplied by Elan Drug Technologies, used in various techniques. Bioz Stars score: 89/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Epitope Biotech 3rt scfv
    Characterization of a novel 3RTau specific single chain antibody. Phage display libraries were positively and negatively panned across recombinant 3RTau and 4RTau protein until a single clone was identified that bound only to 3RTau and did not bind 4RTau. (a) ELISA assay with increasing concentrations of the <t>3RT</t> antibody against immobilized 3R or 4RTau protein (2 μg/ml) EC 50 = 0.064nM. (b) Immunoblot confirmed the specificity and sensitivity of the 3RT antibody (0.1 μg/ml) for 3RTau protein. Further confirmation of specificity was performed by immunohistochemistry analysis with an antibody against the V5 epitope tag on the <t>scFv</t> with (c) N2A neuronal cells infected lentivirus expressing 3RTau or 4RTau, (d) 3RTau tg, 4RTau tg (PS19) or non-tg mouse brain sections, and (e) sections from control, Alzheimer’s, Pick’s, and Progressive Supranuclear Palsy disease patients. The control scFV has no known binding to human or mouse tissues.
    3rt Scfv, supplied by Epitope Biotech, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexa fluor secondary antibody
    Characterization of a novel 3RTau specific single chain antibody. Phage display libraries were positively and negatively panned across recombinant 3RTau and 4RTau protein until a single clone was identified that bound only to 3RTau and did not bind 4RTau. (a) ELISA assay with increasing concentrations of the <t>3RT</t> antibody against immobilized 3R or 4RTau protein (2 μg/ml) EC 50 = 0.064nM. (b) Immunoblot confirmed the specificity and sensitivity of the 3RT antibody (0.1 μg/ml) for 3RTau protein. Further confirmation of specificity was performed by immunohistochemistry analysis with an antibody against the V5 epitope tag on the <t>scFv</t> with (c) N2A neuronal cells infected lentivirus expressing 3RTau or 4RTau, (d) 3RTau tg, 4RTau tg (PS19) or non-tg mouse brain sections, and (e) sections from control, Alzheimer’s, Pick’s, and Progressive Supranuclear Palsy disease patients. The control scFV has no known binding to human or mouse tissues.
    Alexa Fluor Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 677 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Carl Zeiss apotome fluorescence microscope
    Characterization of a novel 3RTau specific single chain antibody. Phage display libraries were positively and negatively panned across recombinant 3RTau and 4RTau protein until a single clone was identified that bound only to 3RTau and did not bind 4RTau. (a) ELISA assay with increasing concentrations of the <t>3RT</t> antibody against immobilized 3R or 4RTau protein (2 μg/ml) EC 50 = 0.064nM. (b) Immunoblot confirmed the specificity and sensitivity of the 3RT antibody (0.1 μg/ml) for 3RTau protein. Further confirmation of specificity was performed by immunohistochemistry analysis with an antibody against the V5 epitope tag on the <t>scFv</t> with (c) N2A neuronal cells infected lentivirus expressing 3RTau or 4RTau, (d) 3RTau tg, 4RTau tg (PS19) or non-tg mouse brain sections, and (e) sections from control, Alzheimer’s, Pick’s, and Progressive Supranuclear Palsy disease patients. The control scFV has no known binding to human or mouse tissues.
    Apotome Fluorescence Microscope, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 92/100, based on 838 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti acetylated α tubulin mouse monoclonal
    Generation of GFAP–Sox4 transgenic mice.  A , Schematic representation of the  GFAP–Sox4  transgene consisting of human GFAP (hGFAP) promoter (positions −2163 to +47), rat Sox4 open reading frame, and IRES-EGFP cassette. Landmark restriction sites in the construct are indicated by letters N ( Nhe I), E ( Eco RI), and A ( Asc I).  B , Southern blot analysis of genomic DNA from wild-type and transgenic mice derived from female founders (#1, #2, #3) after  Eco RI digestion. The 3.4 kb band is indicative of the wild-type  Sox4  gene; the 2.0 kb band represents the  Sox4  transgene. Copy numbers were determined using a phosphorimager and are given below the lanes.  C , Quantitative RT-PCR. Amounts of Sox4 transcripts were determined in total RNA prepared from forebrain and cerebellum of wild-type mice and progeny of transgenic founders at P15. Values were normalized to β-actin for each sample and expressed as multiples of the wild-type values. Error bars indicate SD.  D , Gross morphology of the brain from wild-type (top) and GFAP–Sox4 transgenic (bottom) mice at P19. The right panels focus on the area of the cerebellum. Note the hydrocephalus and the strongly reduced foliation of the transgenic cerebellum.  E , Nissl staining of sections from the mesencephalic region of wild-type and GFAP–Sox4 transgenic brains at P3. Inlays show a magnification of the boxed areas for better visualization of the mesencephalic aqueduct.  F , Immunohistochemical staining of the choroid plexus of wild-type and GFAP–Sox4 transgenic mice at P15 using antibodies directed against Sox9.  G , Confocal imaging of ependymal cilia in wild-type and GFAP–Sox4 transgenic mice at P9 after immunohistochemical staining with antibodies directed against acetylated α tubulin. The same magnifications were used for pictures of wild-type and transgenic cerebella. Scale bars:  E , 1 mm (overview), 50 μm (inlays);  F , 50 μm;  G , 10 μm. wt, Wild type; tg, transgenic.
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    Millipore antibodies against s100
    Trafficking of AFF 1-induced antibodies into the CNS of MBP-α-syn tg mice. (A) Monoclonal AFF 1-induced antibodies were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Alexa-488 tagged mAb-AFF 1 bounded α-syn within cell bodies (arrow-head) and blood vessels (bv). As negative control, a non-immune Alexa-488-tagged IgG1 was used. Scale bar = 5 μm (B) mAb-AFF 1 or non-immune IgG1 were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Time course analysis was performed every 24 h for 3 days, and fluorescence was only increased in brain sections of MBP-α-syn tg animals injected with Alexa-488-tagged mAb-AFF 1. Results are shown as corrected intensity values and expressed as average ± SEM. n = 3 animals per group and time point (C) AFF 1-induced antibodies were detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against Iba1 (microglia) or <t>S100</t> (astrocytes) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in microglial cell bodies and projections, but not in astroglial cells (arrows) (D) AFF 1-induced antibodies detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against p25 (oligodendrocytes) or NeuN (neurons) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in oligodendroglial cell bodies, but not in neurons (arrows). Scale bar = 5 μm.
    Antibodies Against S100, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti laminin
    Photomicrographs illustrating the vascular lesion appearing in the CA3 stratum lacunosum-moleculare after status epilepticus (SE), in pilocarpine-treated rats. The lesion was investigated using an antibody to <t>laminin,</t> which identifies the basal lamina in blood vessels in control tissue (A). Laminin immunoreactivity is markedly upregulated in the damaged area of a pilocarpine-treated rat of the saline-treated group (B) sacrificed 4 days after SE. Pretreatment with the GH secretagogue ghrelin (C) did not affect the increase of laminin immunoreactivity. Notably, pretreatment with JMV-1843 (D) prevented the changes observed in the other treatment groups, which are quantified in E. ## = P
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    Millipore anti rabbit igg h l highly cross adsorbed texas red antibody
    Photomicrographs illustrating the vascular lesion appearing in the CA3 stratum lacunosum-moleculare after status epilepticus (SE), in pilocarpine-treated rats. The lesion was investigated using an antibody to <t>laminin,</t> which identifies the basal lamina in blood vessels in control tissue (A). Laminin immunoreactivity is markedly upregulated in the damaged area of a pilocarpine-treated rat of the saline-treated group (B) sacrificed 4 days after SE. Pretreatment with the GH secretagogue ghrelin (C) did not affect the increase of laminin immunoreactivity. Notably, pretreatment with JMV-1843 (D) prevented the changes observed in the other treatment groups, which are quantified in E. ## = P
    Anti Rabbit Igg H L Highly Cross Adsorbed Texas Red Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM anti iba1
    Photomicrographs illustrating the vascular lesion appearing in the CA3 stratum lacunosum-moleculare after status epilepticus (SE), in pilocarpine-treated rats. The lesion was investigated using an antibody to <t>laminin,</t> which identifies the basal lamina in blood vessels in control tissue (A). Laminin immunoreactivity is markedly upregulated in the damaged area of a pilocarpine-treated rat of the saline-treated group (B) sacrificed 4 days after SE. Pretreatment with the GH secretagogue ghrelin (C) did not affect the increase of laminin immunoreactivity. Notably, pretreatment with JMV-1843 (D) prevented the changes observed in the other treatment groups, which are quantified in E. ## = P
    Anti Iba1, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 94/100, based on 2876 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti sox2
    Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, <t>Sox2</t> ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.
    Anti Sox2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1092 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore monoclonal anti cnpase antibody
    Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, <t>Sox2</t> ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.
    Monoclonal Anti Cnpase Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 233 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore pfa
    Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, <t>Sox2</t> ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.
    Pfa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 14667 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Epitope Biotech rabbit polyclonal antibody
    Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, <t>Sox2</t> ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.
    Rabbit Polyclonal Antibody, supplied by Epitope Biotech, used in various techniques. Bioz Stars score: 92/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in MBP-α-syn tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Hypothesis of the effects of treatment with LV-NR-R80Q-apoB in MBP-α-syn tg mice. In the MBP-α-syn tg model of MSA, oligodendrocytes express high levels of α-syn and release it to the extracellular environment, where it can propagate to astroglial cells or be taken up by microglial cells for degradation. The modified, brain-targeted fusion protein NR-R80Q-apoB (neurosin-apoB) is able to access and degrade extracellular α-syn, as well as be internalized by astroglia and microglia, potentially degrading also intracellular α-syn in those cell types. In LV-Control-treated tg mice ( a ), α-syn propagates and accumulates within astroglial cells. However in LV-NR-R80Q-apoB-treated tg animals neurosin degrades extracellular α-syn ( b ), thus stimulating further release of α-syn from oligodendroglial cells (“sink effect”), reducing its propagation and accumulation in astroglia, and inducing its uptake and degradation by microglia

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Modification

    Delivery of LV-NR-R80Q-apoB reduces the propagation of α-syn to astrocytes and clearance via microglia in MBP-α-syn tg mice. Vibratome brain sections from the non-tg and MBP-α-syn tg that received i.p. injections of LV-Control or LV-NR-R80Q-apoB were double immunofluorescence labeled with antibodies against cellular markers and human α-syn and analyzed with the laser scanning confocal microscope with an optical image of 1 μm with fluorescent signals in co-registry. Dotted box to the left depicts the image field zoomed represented under detail. a Double immunolabeling for the astrocyte marker S100 (red) and human α-syn (green) with nuclei (DAPI, blue). Co-immunolabeling is represented by signal in yellow. b Computer aided image analysis of the % of S100 cells displaying α-syn immunofluorescence in the corpus callosum and striatum. c Double immunolabeling for the microglial marker Iba-1 (red) and human α-syn (green) with nuclei (DAPI, blue). Co-immunolabeling is represented by signal in yellow. d Computer aided image analysis of the % of Iba-1 cells displaying α-syn immunofluorescence in the corpus callosum and striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = 10 μm, detail = 20 μm. # indicates statistical significance ( p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Delivery of LV-NR-R80Q-apoB reduces the propagation of α-syn to astrocytes and clearance via microglia in MBP-α-syn tg mice. Vibratome brain sections from the non-tg and MBP-α-syn tg that received i.p. injections of LV-Control or LV-NR-R80Q-apoB were double immunofluorescence labeled with antibodies against cellular markers and human α-syn and analyzed with the laser scanning confocal microscope with an optical image of 1 μm with fluorescent signals in co-registry. Dotted box to the left depicts the image field zoomed represented under detail. a Double immunolabeling for the astrocyte marker S100 (red) and human α-syn (green) with nuclei (DAPI, blue). Co-immunolabeling is represented by signal in yellow. b Computer aided image analysis of the % of S100 cells displaying α-syn immunofluorescence in the corpus callosum and striatum. c Double immunolabeling for the microglial marker Iba-1 (red) and human α-syn (green) with nuclei (DAPI, blue). Co-immunolabeling is represented by signal in yellow. d Computer aided image analysis of the % of Iba-1 cells displaying α-syn immunofluorescence in the corpus callosum and striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = 10 μm, detail = 20 μm. # indicates statistical significance ( p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Immunofluorescence, Labeling, Microscopy, Immunolabeling, Marker

    Treatment with LV-NR-R80Q-apoB improves the behavioral deficits in MBP-α-syn tg mice. Non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB were tested in the open field test for 4 successive days or for a fixed period to ascertain the functional effects of LV-NR-R80Q-apoB treatment. a Total activity after consecutive days was performed, this is a test of habituation to a novel environment and of memory acquisition. b Retention score analysis representing the ratio between trials 3 and 4. c Total spontaneous activity after 10 min in the open field test. d Total activity in the center of the cage, no differences were noted among the 4 groups. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = 10 μm. * indicates statistical significance ( p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Treatment with LV-NR-R80Q-apoB improves the behavioral deficits in MBP-α-syn tg mice. Non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB were tested in the open field test for 4 successive days or for a fixed period to ascertain the functional effects of LV-NR-R80Q-apoB treatment. a Total activity after consecutive days was performed, this is a test of habituation to a novel environment and of memory acquisition. b Retention score analysis representing the ratio between trials 3 and 4. c Total spontaneous activity after 10 min in the open field test. d Total activity in the center of the cage, no differences were noted among the 4 groups. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = 10 μm. * indicates statistical significance ( p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Functional Assay, Activity Assay

    Delivery of LV-NR-R80Q-apoB ameliorates the neurodegenerative pathology in MBP-α-syn tg mice. Serial longitudinal vibratome sections were analyzed by bright field microscopy. Top row represents a low power overview (40X) of the section and panels in the bottom row show a higher magnification view (400X) of the striatum. a Immunohistochemical analysis with an antibody against the neuronal marker NeuN in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. b , c Stereological analysis using the dissector method to estimate the total numbers of NeuN-positive cells in the frontal cortex and striatum. d Immunohistochemical analysis with an antibody against the astrocyte marker GFAP in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. e , f Semiquantitative analysis of levels of GFAP immunostaining by optical density in the corpus callosum and striatum. g Immunohistochemical analysis with an antibody against the microglial marker Iba-1 in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. h , i Stereological analysis using the dissector method to estimate the total numbers of Iba-1 positive cells in the frontal cortex and striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = in the lower magnification panels 250 μm and in the higher magnification panels 50 μm. * indicates statistical significance ( p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Delivery of LV-NR-R80Q-apoB ameliorates the neurodegenerative pathology in MBP-α-syn tg mice. Serial longitudinal vibratome sections were analyzed by bright field microscopy. Top row represents a low power overview (40X) of the section and panels in the bottom row show a higher magnification view (400X) of the striatum. a Immunohistochemical analysis with an antibody against the neuronal marker NeuN in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. b , c Stereological analysis using the dissector method to estimate the total numbers of NeuN-positive cells in the frontal cortex and striatum. d Immunohistochemical analysis with an antibody against the astrocyte marker GFAP in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. e , f Semiquantitative analysis of levels of GFAP immunostaining by optical density in the corpus callosum and striatum. g Immunohistochemical analysis with an antibody against the microglial marker Iba-1 in brain sections of non-tg mice or MBP-α-syn tg mice injected with LV-Control, or LV-NR-R80Q-apoB vector. h , i Stereological analysis using the dissector method to estimate the total numbers of Iba-1 positive cells in the frontal cortex and striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. Scale bar = in the lower magnification panels 250 μm and in the higher magnification panels 50 μm. * indicates statistical significance ( p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Microscopy, Immunohistochemistry, Marker, Injection, Plasmid Preparation, Immunostaining

    Delivery of LV-NR-R80Q-apoB reduces demyelination in MBP-α-syn tg mice. a Vibratome brain sections were stained with luxol fast blue (LFB) and analyzed by bright field microscopy. The panel in the top is a lower magnification view (40X) showing myelin in blue in white matter tracks. The bottom panels are higher magnification (400X) of the corpus callosum of non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB. Scale bars = Overview 250 μm, corpus callosum 50 μm. b Semiquantitative analysis of LFB staining by optical density in the corpus callosum. c Transmission electron microscopy (TEM) images of myelin sheaths in the corpus callosum of non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB. Representative micrographs taken with TEM are shown at low magnification (5,000x) and high magnification (25,000x). Scale bars = Overview 2.5 μm, Corpus callosum 500 nm. d Computer aided image analysis of the number of myelinated axons in corpus callosum. e Tissue blocks containing the corpus callosum and striatum were fractioned by ultracentrifugation and analyzed by immunoblot with antibodies against MBP and CNP. ( f ) Densitometric analysis of the MBP and CNP signal normalized to the actin signal. * indicates statistical significance ( p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Delivery of LV-NR-R80Q-apoB reduces demyelination in MBP-α-syn tg mice. a Vibratome brain sections were stained with luxol fast blue (LFB) and analyzed by bright field microscopy. The panel in the top is a lower magnification view (40X) showing myelin in blue in white matter tracks. The bottom panels are higher magnification (400X) of the corpus callosum of non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB. Scale bars = Overview 250 μm, corpus callosum 50 μm. b Semiquantitative analysis of LFB staining by optical density in the corpus callosum. c Transmission electron microscopy (TEM) images of myelin sheaths in the corpus callosum of non-tg mice or MBP-α-syn tg mice treated with LV-Control or LV-NR-R80Q-apoB. Representative micrographs taken with TEM are shown at low magnification (5,000x) and high magnification (25,000x). Scale bars = Overview 2.5 μm, Corpus callosum 500 nm. d Computer aided image analysis of the number of myelinated axons in corpus callosum. e Tissue blocks containing the corpus callosum and striatum were fractioned by ultracentrifugation and analyzed by immunoblot with antibodies against MBP and CNP. ( f ) Densitometric analysis of the MBP and CNP signal normalized to the actin signal. * indicates statistical significance ( p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Staining, Microscopy, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy

    Lentivirus vectors over-expressing mutant neurosin reduce the accumulation and propagation of α-synuclein. Wild-type neurosin (LV-wt-NR), neurosin-apoB (LV-wt-NR-apoB), the point mutant neurosin (LV-NR-R80Q) and R80Q-apoB (LV-NR-R80Q-apoB) were cloned into the 3 rd generation lentivirus vector. a B103 neuronal cells were transduced with the vectors and examined by immunoblot for neurosin and the epitope tag V5. b B103 transduced cells were also immunostained for neurosin expression. c Compared to LV-Control, neuronal cells transduced with the neurosin constructs expressed higher levels of neurosin, levels were comparable between the wt and mutant neurosin constructs. d An in vitro neuronal co-culture system was devised to mimic the propagation of α-syn. B103 neuronal “Donor cells” infected with LV-α-syn or LV-Control (red) were plated in cell culture inserts containing a 0.4 μm membrane. B103 neuronal “Acceptor cells” infected with LV-NR-R80Q, LV-NR-R80Q-apoB or LV-Control were plated on coverslips. e Confocal microscopy and immunocytochemical analysis showing α-syn (red) from the Donor Cells was secreted and taken up by Acceptor cells that were double labeled with an antibody against neurosin (green). f Image analysis of double labeled neuronal cells, results are expressed as percent of acceptor cells containing α-syn immunoreactivity. * - indicates one way ANOVA with post hoc Dunnett’s, p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Lentivirus vectors over-expressing mutant neurosin reduce the accumulation and propagation of α-synuclein. Wild-type neurosin (LV-wt-NR), neurosin-apoB (LV-wt-NR-apoB), the point mutant neurosin (LV-NR-R80Q) and R80Q-apoB (LV-NR-R80Q-apoB) were cloned into the 3 rd generation lentivirus vector. a B103 neuronal cells were transduced with the vectors and examined by immunoblot for neurosin and the epitope tag V5. b B103 transduced cells were also immunostained for neurosin expression. c Compared to LV-Control, neuronal cells transduced with the neurosin constructs expressed higher levels of neurosin, levels were comparable between the wt and mutant neurosin constructs. d An in vitro neuronal co-culture system was devised to mimic the propagation of α-syn. B103 neuronal “Donor cells” infected with LV-α-syn or LV-Control (red) were plated in cell culture inserts containing a 0.4 μm membrane. B103 neuronal “Acceptor cells” infected with LV-NR-R80Q, LV-NR-R80Q-apoB or LV-Control were plated on coverslips. e Confocal microscopy and immunocytochemical analysis showing α-syn (red) from the Donor Cells was secreted and taken up by Acceptor cells that were double labeled with an antibody against neurosin (green). f Image analysis of double labeled neuronal cells, results are expressed as percent of acceptor cells containing α-syn immunoreactivity. * - indicates one way ANOVA with post hoc Dunnett’s, p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Expressing, Mutagenesis, Clone Assay, Plasmid Preparation, Transduction, Construct, In Vitro, Co-Culture Assay, Infection, Cell Culture, Confocal Microscopy, Labeling

    Neurosin distribution in the brain following systemic administration of LV-Neurosin-apoB vector into MBP-α-syn tg mice. Non-tg and MBP-α-syn tg mice received a single intra-peritoneal injection of LV-Control or LV-NR-R80Q-apoB (1 × 10 9 TDU). Three months later mice were sacrificed, whole blood, CSF and brains were removed with half frozen for protein analysis and half fixed for tissue section immunohistochemistry. a Representative immunoblot from brain homogenates that were fractioned into soluble and insoluble fractions by ultracentrifugation and analyzed with antibodies for neurosin and the epitope tag, V5. Neurosin was detected as a double band at approximately 28 kDa. b , c Densitometry analysis of the levels of neurosin and V5 immunoreactivity plotted against the actin signal. d Hemibrains were serially sectioned on the longitudinal axis and immunostained with an antibody against neurosin. e Semiquantitative analysis of levels of neurosin immunostaining in the striatum expressed as optical density. f Representative immunoblot analysis of neurosin in plasma portion of the blood and in whole CSF. ( g , h ) Densitometry analysis of neurosin in the blood and CSF. n = 10 mice per group 9–10 m/o at the end of the treatment. * - indicates one way ANOVA with post hoc Dunnett’s, p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Neurosin distribution in the brain following systemic administration of LV-Neurosin-apoB vector into MBP-α-syn tg mice. Non-tg and MBP-α-syn tg mice received a single intra-peritoneal injection of LV-Control or LV-NR-R80Q-apoB (1 × 10 9 TDU). Three months later mice were sacrificed, whole blood, CSF and brains were removed with half frozen for protein analysis and half fixed for tissue section immunohistochemistry. a Representative immunoblot from brain homogenates that were fractioned into soluble and insoluble fractions by ultracentrifugation and analyzed with antibodies for neurosin and the epitope tag, V5. Neurosin was detected as a double band at approximately 28 kDa. b , c Densitometry analysis of the levels of neurosin and V5 immunoreactivity plotted against the actin signal. d Hemibrains were serially sectioned on the longitudinal axis and immunostained with an antibody against neurosin. e Semiquantitative analysis of levels of neurosin immunostaining in the striatum expressed as optical density. f Representative immunoblot analysis of neurosin in plasma portion of the blood and in whole CSF. ( g , h ) Densitometry analysis of neurosin in the blood and CSF. n = 10 mice per group 9–10 m/o at the end of the treatment. * - indicates one way ANOVA with post hoc Dunnett’s, p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Plasmid Preparation, Mouse Assay, Injection, Immunohistochemistry, Immunostaining

    Delivery of LV-NR-R80Q-apoB reduces α-syn accumulation in MBP-α-syn tg mice. a Bright field microscopy analysis of serial longitudinal vibratome sections from the non-tg and MBP-α-syn tg mice immunostained with an antibody against total α-syn (syn-1) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. b Computer aided image analysis for numbers of α-syn positive cells in the striatum and the corpus. c Samples that included the cortex, corpus callosum and striatum were fractioned by ultracentrifugation and analyzed by immunoblot analysis with an antibody against total α-syn (syn-1). d Densitometry analysis of the levels of α-syn immunoreactivity plotted against actin. e Laser scanning confocal microscopy of vibratome sections from the non-tg and MBP-α-syn tg immunostained with an antibody against human α-syn (syn211) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. f Computer aided image analysis of the number of α-syn positive cells in the striatum and the corpus callosum. g Bright field microscopy analysis of serial longitudinal vibratome sections form the non-tg and MBP-α-syn tg immunostained with an antibody against phosphorylated α-syn (Ser129) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. h Computer aided image analysis for numbers of phosphorylated α-syn positive cells in the striatum and the corpus callosum. Scale bars – top panels (low power, 40X) = 250 μm, bottom panel (high power, 400X) = 50 μm. * indicates statistical significance ( p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Delivery of LV-NR-R80Q-apoB reduces α-syn accumulation in MBP-α-syn tg mice. a Bright field microscopy analysis of serial longitudinal vibratome sections from the non-tg and MBP-α-syn tg mice immunostained with an antibody against total α-syn (syn-1) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. b Computer aided image analysis for numbers of α-syn positive cells in the striatum and the corpus. c Samples that included the cortex, corpus callosum and striatum were fractioned by ultracentrifugation and analyzed by immunoblot analysis with an antibody against total α-syn (syn-1). d Densitometry analysis of the levels of α-syn immunoreactivity plotted against actin. e Laser scanning confocal microscopy of vibratome sections from the non-tg and MBP-α-syn tg immunostained with an antibody against human α-syn (syn211) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. f Computer aided image analysis of the number of α-syn positive cells in the striatum and the corpus callosum. g Bright field microscopy analysis of serial longitudinal vibratome sections form the non-tg and MBP-α-syn tg immunostained with an antibody against phosphorylated α-syn (Ser129) and analyzed for α-syn positive cells following treatment with LV-Control, or LV-NR-R80Q-apoB vector. h Computer aided image analysis for numbers of phosphorylated α-syn positive cells in the striatum and the corpus callosum. Scale bars – top panels (low power, 40X) = 250 μm, bottom panel (high power, 400X) = 50 μm. * indicates statistical significance ( p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Microscopy, Plasmid Preparation, Confocal Microscopy

    Cellular uptake of neurosin-apoB in the CNS following systemic delivery of the lentivector to MBP-a-syn tg mice. Non-tg and MBP-α-syn tg received a single i.p. injection with either LV-Control or LV-NR-R80Q-apoB. Three months later mice were sacrificed and brains were analyzed by double immunofluorescence and confocal microscopy for analysis of the co-localization of neurosin with cellular markers. Dotted box to the left depicts the image field zoomed represented under detail. a Double immunolabeling for V5 tag to identify neurosin (red) and the neuronal marker NeuN (green). Co-immunolabeling is represented by signal in yellow. b Computer aided image analysis of the % of NeuN cells displaying neurosin (V5). c Double immunolabeling for V5 tag to identify neurosin (red) and the microglial cell marker Iba-1 (green). d Computer aided image analysis of the % of microglial cells displaying neurosin (V5) immunostaining. e Double immunolabeling for V5 tag to identify neurosin (red) and the astrocytic marker GFAP (green). f Computer aided image analysis of the % of astroglial cells displaying neurosin (V5) immunostaining. g Double immunolabeling for V5 tag to identify neurosin (red) and the oligodendrocyte marker Olig-2 (green). h Computer aided image analysis of the % of oligodendroglial cells displaying neurosin (V5) immunostaining. All images are from the striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. * - indicates one way ANOVA post hoc Dunnett’s p

    Journal: Molecular Neurodegeneration

    Article Title: A brain-targeted, modified neurosin (kallikrein-6) reduces α-synuclein accumulation in a mouse model of multiple system atrophy

    doi: 10.1186/s13024-015-0043-6

    Figure Lengend Snippet: Cellular uptake of neurosin-apoB in the CNS following systemic delivery of the lentivector to MBP-a-syn tg mice. Non-tg and MBP-α-syn tg received a single i.p. injection with either LV-Control or LV-NR-R80Q-apoB. Three months later mice were sacrificed and brains were analyzed by double immunofluorescence and confocal microscopy for analysis of the co-localization of neurosin with cellular markers. Dotted box to the left depicts the image field zoomed represented under detail. a Double immunolabeling for V5 tag to identify neurosin (red) and the neuronal marker NeuN (green). Co-immunolabeling is represented by signal in yellow. b Computer aided image analysis of the % of NeuN cells displaying neurosin (V5). c Double immunolabeling for V5 tag to identify neurosin (red) and the microglial cell marker Iba-1 (green). d Computer aided image analysis of the % of microglial cells displaying neurosin (V5) immunostaining. e Double immunolabeling for V5 tag to identify neurosin (red) and the astrocytic marker GFAP (green). f Computer aided image analysis of the % of astroglial cells displaying neurosin (V5) immunostaining. g Double immunolabeling for V5 tag to identify neurosin (red) and the oligodendrocyte marker Olig-2 (green). h Computer aided image analysis of the % of oligodendroglial cells displaying neurosin (V5) immunostaining. All images are from the striatum. n = 10 mice per group 9–10 m/o at the end of the treatment. * - indicates one way ANOVA post hoc Dunnett’s p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia), Iba1 (microglia), Olig-2 (oligodendrocytes), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with FITC-tagged secondary antibody (Vector Laboratories, 1:75) and the V5 tagged neurosin with the Tyramide Signal Amplification™-Direct system (NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Injection, Immunofluorescence, Confocal Microscopy, Immunolabeling, Marker, Immunostaining

    Delivery of the 3RT-apoB antibody reduces neurodegeneration in the 3RTau tg model. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. The neocortex and hippocampus were immunostained with antibodies against the (a) astrocyte glial fibrillary acidic protein (GFAP), (c) neuronal marker NeuN, (e) and dendritic marker MAP2 (green). (b) GFAP immunoreactivity expressed as corrected optical density of sections. (d) Stereological estimates (dissector method) of total NeuN-positive neuronal counts in the frontal cortex. (f) Image analysis of % area of the neuropil covered by MAP2 post-synaptic processes. * indicates statistical significance p

    Journal: Acta neuropathologica

    Article Title: Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders

    doi: 10.1007/s00401-018-1869-0

    Figure Lengend Snippet: Delivery of the 3RT-apoB antibody reduces neurodegeneration in the 3RTau tg model. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. The neocortex and hippocampus were immunostained with antibodies against the (a) astrocyte glial fibrillary acidic protein (GFAP), (c) neuronal marker NeuN, (e) and dendritic marker MAP2 (green). (b) GFAP immunoreactivity expressed as corrected optical density of sections. (d) Stereological estimates (dissector method) of total NeuN-positive neuronal counts in the frontal cortex. (f) Image analysis of % area of the neuropil covered by MAP2 post-synaptic processes. * indicates statistical significance p

    Article Snippet: To determine the co-localization between 3RT scFv and cells in the CNS, 40 μm-thick vibratome sections were immunolabeled with the rabbit polyclonal antibody against the 3RT scFv (V5 epitope) and Map2 (neuronal dendritic marker, Chemicon), NeuN (neuronal nuclear marker, Millipore), GFAP (astrocyte marker, Chemicon) or Iba1 (microglia marker, Wako).

    Techniques: Mouse Assay, Injection, Marker

    The 3RT-apoB antibody co-localizes with neurons in the brain of 3RTau tg mice. Representative images of double immunocytochemical labeling in the neocortex of non-tg and 3RTau tg mice treated with LV-3RT, LV-3RT-apoB and LV-control with antibodies to V5 (red, 3RT epitope tag) and the neuronal markers (a) NeuN or (c) MAP2, astrocyte marker (e) GFAP, and microglia marker (g) Iba1 imaged analyzed with the laser scanning confocal microscope. Analysis of % cells showing co-localization between V5 (3RT) and (b) NeuN, (d) MAP2, (f) GFAP, and (h) Iba1. Scale bar represents 10μm, and 5μm in detail figure. For analysis, 3RTau tg LV-control (n=12); LV-3RT (n=12); LV-3RT-apoB (n=12) and non-tg mice LV-control (n=6).

    Journal: Acta neuropathologica

    Article Title: Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders

    doi: 10.1007/s00401-018-1869-0

    Figure Lengend Snippet: The 3RT-apoB antibody co-localizes with neurons in the brain of 3RTau tg mice. Representative images of double immunocytochemical labeling in the neocortex of non-tg and 3RTau tg mice treated with LV-3RT, LV-3RT-apoB and LV-control with antibodies to V5 (red, 3RT epitope tag) and the neuronal markers (a) NeuN or (c) MAP2, astrocyte marker (e) GFAP, and microglia marker (g) Iba1 imaged analyzed with the laser scanning confocal microscope. Analysis of % cells showing co-localization between V5 (3RT) and (b) NeuN, (d) MAP2, (f) GFAP, and (h) Iba1. Scale bar represents 10μm, and 5μm in detail figure. For analysis, 3RTau tg LV-control (n=12); LV-3RT (n=12); LV-3RT-apoB (n=12) and non-tg mice LV-control (n=6).

    Article Snippet: To determine the co-localization between 3RT scFv and cells in the CNS, 40 μm-thick vibratome sections were immunolabeled with the rabbit polyclonal antibody against the 3RT scFv (V5 epitope) and Map2 (neuronal dendritic marker, Chemicon), NeuN (neuronal nuclear marker, Millipore), GFAP (astrocyte marker, Chemicon) or Iba1 (microglia marker, Wako).

    Techniques: Mouse Assay, Labeling, Marker, Microscopy

    PGRN overexpression in hippocampus decreases amyloid plaque load in 5xFAD mice. ( a ) Representative images of the contralateral and ipsilateral hemispheres of Lenti-Ctrl- and Lenti-PGRN-injected 5xFAD mice. Dashed lines indicate boundaries used for quantification of dentate gyrus (DG) and non-dentate gyrus hippocampal (non-DG HP) areas. PGRN (red), 3D6 + plaques (green), and ThioS + plaques (magenta). Scale bar, 200 μm. ( b ) Quantification of PGRN immunostaining normalized to the uninjected contralateral hemisphere for Lenti-Ctrl- or Lenti-PGRN-injected mice. Two Lenti-PGRN-injected mice lacking sufficient overexpression (i.e.

    Journal: Nature medicine

    Article Title: Progranulin Protects against Amyloid β Deposition and Toxicity in Alzheimer’s Disease Mouse Models

    doi: 10.1038/nm.3672

    Figure Lengend Snippet: PGRN overexpression in hippocampus decreases amyloid plaque load in 5xFAD mice. ( a ) Representative images of the contralateral and ipsilateral hemispheres of Lenti-Ctrl- and Lenti-PGRN-injected 5xFAD mice. Dashed lines indicate boundaries used for quantification of dentate gyrus (DG) and non-dentate gyrus hippocampal (non-DG HP) areas. PGRN (red), 3D6 + plaques (green), and ThioS + plaques (magenta). Scale bar, 200 μm. ( b ) Quantification of PGRN immunostaining normalized to the uninjected contralateral hemisphere for Lenti-Ctrl- or Lenti-PGRN-injected mice. Two Lenti-PGRN-injected mice lacking sufficient overexpression (i.e.

    Article Snippet: For immunofluorescence staining, sections were incubated with anti-Iba1 (#019–19741, 1:500, Wako, Osaka, Japan), 3D6 (5 μg/ml, Elan), anti-NeuN (MAB377, 1:500, Millipore), or anti-MAP2 (MAB3418, 1:300, Millipore), Immunoreactive structures were detected with Alexa Fluor donkey anti-mouse or rabbit secondary antibodies in the 405-, 488-, 555-, or 647-nm range (Invitrogen, Carlsbad, CA).

    Techniques: Over Expression, Mouse Assay, Injection, Immunostaining

    Characterization of a novel 3RTau specific single chain antibody. Phage display libraries were positively and negatively panned across recombinant 3RTau and 4RTau protein until a single clone was identified that bound only to 3RTau and did not bind 4RTau. (a) ELISA assay with increasing concentrations of the 3RT antibody against immobilized 3R or 4RTau protein (2 μg/ml) EC 50 = 0.064nM. (b) Immunoblot confirmed the specificity and sensitivity of the 3RT antibody (0.1 μg/ml) for 3RTau protein. Further confirmation of specificity was performed by immunohistochemistry analysis with an antibody against the V5 epitope tag on the scFv with (c) N2A neuronal cells infected lentivirus expressing 3RTau or 4RTau, (d) 3RTau tg, 4RTau tg (PS19) or non-tg mouse brain sections, and (e) sections from control, Alzheimer’s, Pick’s, and Progressive Supranuclear Palsy disease patients. The control scFV has no known binding to human or mouse tissues.

    Journal: Acta neuropathologica

    Article Title: Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders

    doi: 10.1007/s00401-018-1869-0

    Figure Lengend Snippet: Characterization of a novel 3RTau specific single chain antibody. Phage display libraries were positively and negatively panned across recombinant 3RTau and 4RTau protein until a single clone was identified that bound only to 3RTau and did not bind 4RTau. (a) ELISA assay with increasing concentrations of the 3RT antibody against immobilized 3R or 4RTau protein (2 μg/ml) EC 50 = 0.064nM. (b) Immunoblot confirmed the specificity and sensitivity of the 3RT antibody (0.1 μg/ml) for 3RTau protein. Further confirmation of specificity was performed by immunohistochemistry analysis with an antibody against the V5 epitope tag on the scFv with (c) N2A neuronal cells infected lentivirus expressing 3RTau or 4RTau, (d) 3RTau tg, 4RTau tg (PS19) or non-tg mouse brain sections, and (e) sections from control, Alzheimer’s, Pick’s, and Progressive Supranuclear Palsy disease patients. The control scFV has no known binding to human or mouse tissues.

    Article Snippet: To determine the co-localization between 3RT scFv and cells in the CNS, 40 μm-thick vibratome sections were immunolabeled with the rabbit polyclonal antibody against the 3RT scFv (V5 epitope) and Map2 (neuronal dendritic marker, Chemicon), NeuN (neuronal nuclear marker, Millipore), GFAP (astrocyte marker, Chemicon) or Iba1 (microglia marker, Wako).

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Infection, Expressing, Binding Assay

    The 3RT-apoB antibody reduces Tau accumulation in the brain. (a) Representative, immunoblot analysis of brain homogenates from non-tg and 3RTau tg mice treated with LV-3RT, LV-3RT-apoB or LV-control showing levels of Tau proteins: 3RTau, p-Tau (PHF1), and total Tau (T-Tau) as well as the level of the 3RT scFv protein (V5) with ß-actin as a loading control. Image analysis of pixels of (b) V5, (c) 3RTau, (d) PHF1, and (e) T-Tau immunoreactivity analyzed as ratio to ß-actin signal. * indicates statistical significance p

    Journal: Acta neuropathologica

    Article Title: Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders

    doi: 10.1007/s00401-018-1869-0

    Figure Lengend Snippet: The 3RT-apoB antibody reduces Tau accumulation in the brain. (a) Representative, immunoblot analysis of brain homogenates from non-tg and 3RTau tg mice treated with LV-3RT, LV-3RT-apoB or LV-control showing levels of Tau proteins: 3RTau, p-Tau (PHF1), and total Tau (T-Tau) as well as the level of the 3RT scFv protein (V5) with ß-actin as a loading control. Image analysis of pixels of (b) V5, (c) 3RTau, (d) PHF1, and (e) T-Tau immunoreactivity analyzed as ratio to ß-actin signal. * indicates statistical significance p

    Article Snippet: To determine the co-localization between 3RT scFv and cells in the CNS, 40 μm-thick vibratome sections were immunolabeled with the rabbit polyclonal antibody against the 3RT scFv (V5 epitope) and Map2 (neuronal dendritic marker, Chemicon), NeuN (neuronal nuclear marker, Millipore), GFAP (astrocyte marker, Chemicon) or Iba1 (microglia marker, Wako).

    Techniques: Mouse Assay

    Systemic delivery of the 3RT antibody with a lentiviral vector reduces the accumulation of 3RTau in the brain of transgenic mice. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. (a,b) Immunocytochemical analysis with antibodies against the V5 tag to detect the 3RT scFv in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (c,d) Immunocytochemical analysis with antibodies against 3RTau in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (e,f) Immunocytochemical analysis with antibodies against phosphor-Tau (PHF-1) in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (g,h) Immunocytochemical analysis with antibodies against total Tau (T-Tau) in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (b,d,f,h) Immunoreactivity semi-quantified and expressed as corrected optical density of sections. * indicates statistical significance p

    Journal: Acta neuropathologica

    Article Title: Selective targeting of 3 repeat Tau with brain penetrating single chain antibodies for the treatment of neurodegenerative disorders

    doi: 10.1007/s00401-018-1869-0

    Figure Lengend Snippet: Systemic delivery of the 3RT antibody with a lentiviral vector reduces the accumulation of 3RTau in the brain of transgenic mice. 4-month-old 3RTau tg and non-tg mice received a single i.p. injection of LV-3RT, LV-3RT-apoB or LV-control and were sacrificed 4 months later for analysis. (a,b) Immunocytochemical analysis with antibodies against the V5 tag to detect the 3RT scFv in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (c,d) Immunocytochemical analysis with antibodies against 3RTau in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (e,f) Immunocytochemical analysis with antibodies against phosphor-Tau (PHF-1) in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (g,h) Immunocytochemical analysis with antibodies against total Tau (T-Tau) in the frontal cortex and hippocampus of non-tg and 3RTau tg mice. (b,d,f,h) Immunoreactivity semi-quantified and expressed as corrected optical density of sections. * indicates statistical significance p

    Article Snippet: To determine the co-localization between 3RT scFv and cells in the CNS, 40 μm-thick vibratome sections were immunolabeled with the rabbit polyclonal antibody against the 3RT scFv (V5 epitope) and Map2 (neuronal dendritic marker, Chemicon), NeuN (neuronal nuclear marker, Millipore), GFAP (astrocyte marker, Chemicon) or Iba1 (microglia marker, Wako).

    Techniques: Plasmid Preparation, Transgenic Assay, Mouse Assay, Injection

    Generation of GFAP–Sox4 transgenic mice.  A , Schematic representation of the  GFAP–Sox4  transgene consisting of human GFAP (hGFAP) promoter (positions −2163 to +47), rat Sox4 open reading frame, and IRES-EGFP cassette. Landmark restriction sites in the construct are indicated by letters N ( Nhe I), E ( Eco RI), and A ( Asc I).  B , Southern blot analysis of genomic DNA from wild-type and transgenic mice derived from female founders (#1, #2, #3) after  Eco RI digestion. The 3.4 kb band is indicative of the wild-type  Sox4  gene; the 2.0 kb band represents the  Sox4  transgene. Copy numbers were determined using a phosphorimager and are given below the lanes.  C , Quantitative RT-PCR. Amounts of Sox4 transcripts were determined in total RNA prepared from forebrain and cerebellum of wild-type mice and progeny of transgenic founders at P15. Values were normalized to β-actin for each sample and expressed as multiples of the wild-type values. Error bars indicate SD.  D , Gross morphology of the brain from wild-type (top) and GFAP–Sox4 transgenic (bottom) mice at P19. The right panels focus on the area of the cerebellum. Note the hydrocephalus and the strongly reduced foliation of the transgenic cerebellum.  E , Nissl staining of sections from the mesencephalic region of wild-type and GFAP–Sox4 transgenic brains at P3. Inlays show a magnification of the boxed areas for better visualization of the mesencephalic aqueduct.  F , Immunohistochemical staining of the choroid plexus of wild-type and GFAP–Sox4 transgenic mice at P15 using antibodies directed against Sox9.  G , Confocal imaging of ependymal cilia in wild-type and GFAP–Sox4 transgenic mice at P9 after immunohistochemical staining with antibodies directed against acetylated α tubulin. The same magnifications were used for pictures of wild-type and transgenic cerebella. Scale bars:  E , 1 mm (overview), 50 μm (inlays);  F , 50 μm;  G , 10 μm. wt, Wild type; tg, transgenic.

    Journal: The Journal of Neuroscience

    Article Title: Prolonged Glial Expression of Sox4 in the CNS Leads to Architectural Cerebellar Defects and Ataxia

    doi: 10.1523/JNEUROSCI.1384-07.2007

    Figure Lengend Snippet: Generation of GFAP–Sox4 transgenic mice. A , Schematic representation of the GFAP–Sox4 transgene consisting of human GFAP (hGFAP) promoter (positions −2163 to +47), rat Sox4 open reading frame, and IRES-EGFP cassette. Landmark restriction sites in the construct are indicated by letters N ( Nhe I), E ( Eco RI), and A ( Asc I). B , Southern blot analysis of genomic DNA from wild-type and transgenic mice derived from female founders (#1, #2, #3) after Eco RI digestion. The 3.4 kb band is indicative of the wild-type Sox4 gene; the 2.0 kb band represents the Sox4 transgene. Copy numbers were determined using a phosphorimager and are given below the lanes. C , Quantitative RT-PCR. Amounts of Sox4 transcripts were determined in total RNA prepared from forebrain and cerebellum of wild-type mice and progeny of transgenic founders at P15. Values were normalized to β-actin for each sample and expressed as multiples of the wild-type values. Error bars indicate SD. D , Gross morphology of the brain from wild-type (top) and GFAP–Sox4 transgenic (bottom) mice at P19. The right panels focus on the area of the cerebellum. Note the hydrocephalus and the strongly reduced foliation of the transgenic cerebellum. E , Nissl staining of sections from the mesencephalic region of wild-type and GFAP–Sox4 transgenic brains at P3. Inlays show a magnification of the boxed areas for better visualization of the mesencephalic aqueduct. F , Immunohistochemical staining of the choroid plexus of wild-type and GFAP–Sox4 transgenic mice at P15 using antibodies directed against Sox9. G , Confocal imaging of ependymal cilia in wild-type and GFAP–Sox4 transgenic mice at P9 after immunohistochemical staining with antibodies directed against acetylated α tubulin. The same magnifications were used for pictures of wild-type and transgenic cerebella. Scale bars: E , 1 mm (overview), 50 μm (inlays); F , 50 μm; G , 10 μm. wt, Wild type; tg, transgenic.

    Article Snippet: For immunohistochemistry, the following primary antibodies were used in various combinations: anti-Sox10 guinea pig antiserum (1:1000 dilution) , anti-acetylated α tubulin mouse monoclonal (1:100 dilution) , anti-NeuN mouse monoclonal (1:500 dilution, catalog #MAB377; Millipore, Temecula, CA), anti-Tuj1 mouse monoclonal (1:5000 dilution, catalog #MMS-435P; Covance, Denver, PA), anti-microtubule-associated protein 2 (MAP2) mouse monoclonal (1:125 dilution, catalog #M1406; Sigma, St. Louis, MO), anti-GFAP mouse monoclonal (1:1000 dilution, catalog #MAB360; Chemicon), anti-green fluorescent protein (GFP) mouse monoclonal (1:100 dilution, catalog #1814460; Roche), anti-GFP rabbit antiserum (1:500 dilution, catalog # ; Invitrogen), anti-brain fatty acid-binding protein (B-FABP) rabbit antiserum (1:10,000 dilution; gift of C. Birchmeier and T. Müller, Max-Delbrück-Centrum, Berlin, Germany), affinity-purified anti-Sox9 rabbit antiserum (1:2000 dilution) , anti-SoxB1 rabbit antiserum (1:500 dilution) , anti-calbindin D28K rabbit antiserum (1:3000 dilution, catalog #CB-38; Swant, Bellinzona, Switzerland), anti-calretinin rabbit antiserum (1:2000 dilution, catalog #7699/4; Swant), anti-Pax2 rabbit antiserum (1:200 dilution, catalog #71-6000; Zymed, San Francisco, CA), anti-Oct6 rabbit antiserum (1:1000 dilution) , anti-Ki67 rabbit monoclonal (1:500 dilution, catalog #RM-9106; LabVision, Fremont, CA), anti-retinoid-related orphan receptor α (RORα) goat antiserum (1:100 dilution, catalog #sc-6062; Santa Cruz Biotechnology, Santa Cruz, CA), and anti-Sox11 guinea pig antiserum (1:1000 dilution; M. Hoser and E. Sock, unpublished).

    Techniques: Transgenic Assay, Mouse Assay, Construct, Southern Blot, Derivative Assay, Quantitative RT-PCR, Staining, Immunohistochemistry, Imaging

    Trafficking of AFF 1-induced antibodies into the CNS of MBP-α-syn tg mice. (A) Monoclonal AFF 1-induced antibodies were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Alexa-488 tagged mAb-AFF 1 bounded α-syn within cell bodies (arrow-head) and blood vessels (bv). As negative control, a non-immune Alexa-488-tagged IgG1 was used. Scale bar = 5 μm (B) mAb-AFF 1 or non-immune IgG1 were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Time course analysis was performed every 24 h for 3 days, and fluorescence was only increased in brain sections of MBP-α-syn tg animals injected with Alexa-488-tagged mAb-AFF 1. Results are shown as corrected intensity values and expressed as average ± SEM. n = 3 animals per group and time point (C) AFF 1-induced antibodies were detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against Iba1 (microglia) or S100 (astrocytes) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in microglial cell bodies and projections, but not in astroglial cells (arrows) (D) AFF 1-induced antibodies detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against p25 (oligodendrocytes) or NeuN (neurons) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in oligodendroglial cell bodies, but not in neurons (arrows). Scale bar = 5 μm.

    Journal: Molecular Neurodegeneration

    Article Title: Active immunization against alpha-synuclein ameliorates the degenerative pathology and prevents demyelination in a model of multiple system atrophy

    doi: 10.1186/s13024-015-0008-9

    Figure Lengend Snippet: Trafficking of AFF 1-induced antibodies into the CNS of MBP-α-syn tg mice. (A) Monoclonal AFF 1-induced antibodies were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Alexa-488 tagged mAb-AFF 1 bounded α-syn within cell bodies (arrow-head) and blood vessels (bv). As negative control, a non-immune Alexa-488-tagged IgG1 was used. Scale bar = 5 μm (B) mAb-AFF 1 or non-immune IgG1 were tagged with Alexa-488 and administered to non-tg or MBP-α-syn mice. Time course analysis was performed every 24 h for 3 days, and fluorescence was only increased in brain sections of MBP-α-syn tg animals injected with Alexa-488-tagged mAb-AFF 1. Results are shown as corrected intensity values and expressed as average ± SEM. n = 3 animals per group and time point (C) AFF 1-induced antibodies were detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against Iba1 (microglia) or S100 (astrocytes) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in microglial cell bodies and projections, but not in astroglial cells (arrows) (D) AFF 1-induced antibodies detected with and FITC-tagged anti-mouse antibody in brain sections of immunized MBP-α-syn tg mice (green), together with an antibody against p25 (oligodendrocytes) or NeuN (neurons) (red). Cell nuclei were stained with DAPI (blue). Colocalization was observed in oligodendroglial cell bodies, but not in neurons (arrows). Scale bar = 5 μm.

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia, Sigma, 1:250), Iba1 (microglia), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with the Tyramide Signal Amplification™-Direct system (1:100, NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma, 1:250) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Negative Control, Fluorescence, Injection, Staining

    Immunization with AFF 1 increases microglial α-syn clearance in MBP-α-syn tg mice. (A) Double immunostaining for the microglial marker Iba1 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of Iba1-positive cells is denoted by arrows and the location of α-syn-positive cells by asterisks. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (B) Quantification of the percentage of colocalization between Iba1 and α-syn (C) Double immunostaining for the oligodendroglial marker p25 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of p25-positive cells is denoted by arrows. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (D) Quantification of the percentage of colocalization between p25 and α-syn (E) Double immunostaining for the astroglial marker S100 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of S100-positive cells is denoted by arrows and the location of α-syn-positive cells by asterisks. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (F) Quantification of the percentage of colocalization between S100 and α-syn (G) Double immunostaining for the neuronal marker NeuN (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. Cell nuclei were stained with DAPI (blue). Scale bar = 5 μm. (H) Quantification of the percentage of colocalization between NeuN and α-syn. Results are expressed as average ± SEM. (*) p

    Journal: Molecular Neurodegeneration

    Article Title: Active immunization against alpha-synuclein ameliorates the degenerative pathology and prevents demyelination in a model of multiple system atrophy

    doi: 10.1186/s13024-015-0008-9

    Figure Lengend Snippet: Immunization with AFF 1 increases microglial α-syn clearance in MBP-α-syn tg mice. (A) Double immunostaining for the microglial marker Iba1 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of Iba1-positive cells is denoted by arrows and the location of α-syn-positive cells by asterisks. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (B) Quantification of the percentage of colocalization between Iba1 and α-syn (C) Double immunostaining for the oligodendroglial marker p25 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of p25-positive cells is denoted by arrows. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (D) Quantification of the percentage of colocalization between p25 and α-syn (E) Double immunostaining for the astroglial marker S100 (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. The location of S100-positive cells is denoted by arrows and the location of α-syn-positive cells by asterisks. Cell nuclei were stained with DAPI (blue). Scale bar = 10 μm. (F) Quantification of the percentage of colocalization between S100 and α-syn (G) Double immunostaining for the neuronal marker NeuN (red) and α-syn (green) in vehicle- and AFF 1-immunized MBP-α-syn tg mice. Cell nuclei were stained with DAPI (blue). Scale bar = 5 μm. (H) Quantification of the percentage of colocalization between NeuN and α-syn. Results are expressed as average ± SEM. (*) p

    Article Snippet: Vibratome sections were immunolabeled with antibodies against S100 (astroglia, Sigma, 1:250), Iba1 (microglia), p25 (oligodendrocytes) or NeuN (neurons), and the immunoreactive structures were detected with the Tyramide Signal Amplification™-Direct system (1:100, NEN Life Sciences), while α-syn was detected with an antibody specific for human α-syn (SYN211, Sigma, 1:250) and a FITC-tagged secondary antibody (Vector Laboratories, 1:75).

    Techniques: Mouse Assay, Double Immunostaining, Marker, Staining

    Photomicrographs illustrating the vascular lesion appearing in the CA3 stratum lacunosum-moleculare after status epilepticus (SE), in pilocarpine-treated rats. The lesion was investigated using an antibody to laminin, which identifies the basal lamina in blood vessels in control tissue (A). Laminin immunoreactivity is markedly upregulated in the damaged area of a pilocarpine-treated rat of the saline-treated group (B) sacrificed 4 days after SE. Pretreatment with the GH secretagogue ghrelin (C) did not affect the increase of laminin immunoreactivity. Notably, pretreatment with JMV-1843 (D) prevented the changes observed in the other treatment groups, which are quantified in E. ## = P

    Journal: PLoS ONE

    Article Title: Protective but Not Anticonvulsant Effects of Ghrelin and JMV-1843 in the Pilocarpine Model of Status epilepticus

    doi: 10.1371/journal.pone.0072716

    Figure Lengend Snippet: Photomicrographs illustrating the vascular lesion appearing in the CA3 stratum lacunosum-moleculare after status epilepticus (SE), in pilocarpine-treated rats. The lesion was investigated using an antibody to laminin, which identifies the basal lamina in blood vessels in control tissue (A). Laminin immunoreactivity is markedly upregulated in the damaged area of a pilocarpine-treated rat of the saline-treated group (B) sacrificed 4 days after SE. Pretreatment with the GH secretagogue ghrelin (C) did not affect the increase of laminin immunoreactivity. Notably, pretreatment with JMV-1843 (D) prevented the changes observed in the other treatment groups, which are quantified in E. ## = P

    Article Snippet: After several washes, sections were blocked in phosphate buffered saline (PBS, pH 7.4) containing 2% goat serum and were respectively incubated with antibodies able to reveal astrocytic , neuronal and vessel wall lesions ; in particular, we used three monoclonal antibodies: anti-GFAP (1∶1000, Sigma-Aldrich); anti-neuron-specific nuclear protein (NeuN, 1∶200; Chemicon, Temecula, CA, USA); anti-laminin (1∶2000, Sigma-Aldrich).

    Techniques:

    Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, Sox2 ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.

    Journal: The Journal of Neuroscience

    Article Title: Sox21 Promotes Hippocampal Adult Neurogenesis via the Transcriptional Repression of the Hes5 Gene

    doi: 10.1523/JNEUROSCI.5803-11.2012

    Figure Lengend Snippet: Specific expression of Sox21 in NS/PCs in the adult DG. Immunohistochemical analyses of Sox21 in the adult DG. A–D , The areas surrounded by white rectangles in the top panels are shown in the magnified views in the middle three panels. Arrows indicate Sox21 and marker double-positive cells; arrowheads indicate Sox21 single-positive cells. The cells highlighted by yellow arrows are also shown using orthogonal views at higher magnification in the bottom panels (top planes are through the x-z axes, and right planes are through the y-z axes). In the DG, Sox21 expression was restricted to cells in the SGZ. Sox21 was expressed in FABP7-positive ( A ) and GFAP-positive ( B ) NS/PCs with horizontally oriented cell bodies typical of type 2a cells and also in GFAP-positive stem-like cells with radial glia-like fibers ( B , asterisk) typical of type 1 cells. C , D , The Sox21 expression pattern showed incomplete concordance with the other NS/PC markers, Sox2 ( C ) and Nestin ( D ). E , Very few Sox21-positive cells coexpressed PSA-NCAM, a marker for immature neurons. Arrowheads indicate Sox21 single-positive cells. F , The Sox21-positive cell population was mutually exclusive of NeuN-positive neurons in the DG. Arrowheads show Sox21 single-positive cells. G , H , Sox21 was not expressed in S100β-positive astrocytes or GSTπ-positive oligodendrocytes in the DG, although some S100β-positive cells outside the GCL (arrowhead) expressed Sox21. I , A schematic summary of neuronal differentiation in the adult DG and the markers expressed at each stage. Scale bars: A–D , top, 100 μm; middle, 50 μm; bottom, 20 μm; E , F (panels with orthogonal views) G , H , 50 μm; E , F (magnified views), 20 μm.

    Article Snippet: The following primary antibodies were used in this study: anti-βIII-tubulin (rabbit IgG; Covance, PRB-435P; 1:500), anti-glial fibrillary acidic protein (GFAP; rabbit IgG; DAKO, Z0334; 1:500), anti-GFAP (mouse IgG; Sigma-Aldrich, G3893; 1:200), anti-fatty acid binding protein 7 (FABP7; rabbit IgG; Millipore, AB9558; 1:250), anti-Sox21 (goat IgG; R & D Systems, AF3538; 1:100), anti-Sox2 (rabbit IgG; Millipore, AB5603; 1:200), anti-Sox1/(2)/3 (rabbit IgG; gift from Dr. H. Kondoh, Osaka University, Osaka, Japan; 1:5000) , anti-Nestin (mouse IgG; BD Biosciences/PharMingen, 556309; 1:200), anti-doublecortin (DCX; goat IgG; Santa Cruz Biotechnology, sc-8066; 1:100), anti- DCX (rabbit IgG; Abcam, ab18723; 1:100), anti-polysialylated neuronal cell adhesion molecule (PSA-NCAM; mouse IgMκ; gift from Dr. T. Seki, Tokyo Medical University, Tokyo, Japan; 1:200), anti-NeuN (mouse IgG; Millipore, MAB377; 1:100), anti-S100β (mouse IgG; Sigma-Aldrich, S2532; 1:200), anti-glutathione S -transferase π (mouse IgG; BD Biosciences/PharMingen, 610719; 1:200), anti-Brn2 (Santa Cruz Biotechnology, sc-6029; 1:100), anti-Tbr1 (rabbit IgG; gift from Dr. R. F. Hevner, University of Washington, Seattle, WA; 1:10,000), anti-Ascl1 (mouse IgG; BD Biosciences/PharMingen, 556604; 1:100), anti-Tbr2 (rabbit IgG; Abcam, ab23345; 1:200), anti-Ki67 (rabbit IgG; Abcam, ab15580; 1:3000), anti-GFP (rabbit IgG, Medical and Biological Laboratories, 598, 1:500; goat IgG, Rockland, 600-101-215, 1:500), anti-BrdU (rat IgG; Abcam, ab6326; 1:100), anti-lacZ (rabbit IgG, Cappel, 55976, 1:500; goat IgG, Cappel, 56028, 1:500), and normal IgG (rabbit IgG, Millipore, DAM1421465; goat IgG, Santa Cruz Biotechnology, sc-2028).

    Techniques: Expressing, Immunohistochemistry, Marker

    Repression of Hes5 gene promoter activity by Sox21 but not Sox2. AHP cells were transfected with Hes5 promoter–luciferase reporter constructs (1 μg per well) with or without expression vectors for NICD , Sox21 , and/or Sox2 . The histogram shows the ratio of firefly luciferase to that in cells transfected with the reporter construct and the NICD -expressing vector. The transfection efficiency was normalized to Renilla luciferase activity. A , B , The Hes5 DRE and proximal promoter (−2767 to +73 bp from TSS) and the proximal promoter alone (−687 to +73 bp) were used as reporter constructs in A and B , respectively. The open columns in A show the relative luciferase activities using the intact reporter construct, and the closed columns reflect the construct mutagenized at the Sox-binding sequence of the DRE (ACAAAGG to AagcttG). NICD -expression vector, 0.1 μg per well; Sox21 -expression vector, 0.0001, 0.001, and 0.01 μg per well. C , The Hes5 DRE (−2676 to −2244 bp) combined with the β-globin minimal promoter was used. The open columns indicate the intact reporter construct, and the closed columns indicate the mutagenized construct as shown in A . NICD, 1 μg per well; Sox21, 0.0001, 0.001, and 0.01 μg per well. D , E , The Hes5 DRE and proximal promoter were used. D , NICD, 0.1 μg per well, Sox2, 0.0001, 0.001, and 0.01 μg per well. E , NICD, 0.1 μg per well; Sox21, 0.01 μg per well; Sox2, 0.0001, 0.001, and 0.01 μg per well. Data represent the mean ± SE. Statistical significance versus the intact promoter was assessed by two-sided t test ( n > 4). ** p

    Journal: The Journal of Neuroscience

    Article Title: Sox21 Promotes Hippocampal Adult Neurogenesis via the Transcriptional Repression of the Hes5 Gene

    doi: 10.1523/JNEUROSCI.5803-11.2012

    Figure Lengend Snippet: Repression of Hes5 gene promoter activity by Sox21 but not Sox2. AHP cells were transfected with Hes5 promoter–luciferase reporter constructs (1 μg per well) with or without expression vectors for NICD , Sox21 , and/or Sox2 . The histogram shows the ratio of firefly luciferase to that in cells transfected with the reporter construct and the NICD -expressing vector. The transfection efficiency was normalized to Renilla luciferase activity. A , B , The Hes5 DRE and proximal promoter (−2767 to +73 bp from TSS) and the proximal promoter alone (−687 to +73 bp) were used as reporter constructs in A and B , respectively. The open columns in A show the relative luciferase activities using the intact reporter construct, and the closed columns reflect the construct mutagenized at the Sox-binding sequence of the DRE (ACAAAGG to AagcttG). NICD -expression vector, 0.1 μg per well; Sox21 -expression vector, 0.0001, 0.001, and 0.01 μg per well. C , The Hes5 DRE (−2676 to −2244 bp) combined with the β-globin minimal promoter was used. The open columns indicate the intact reporter construct, and the closed columns indicate the mutagenized construct as shown in A . NICD, 1 μg per well; Sox21, 0.0001, 0.001, and 0.01 μg per well. D , E , The Hes5 DRE and proximal promoter were used. D , NICD, 0.1 μg per well, Sox2, 0.0001, 0.001, and 0.01 μg per well. E , NICD, 0.1 μg per well; Sox21, 0.01 μg per well; Sox2, 0.0001, 0.001, and 0.01 μg per well. Data represent the mean ± SE. Statistical significance versus the intact promoter was assessed by two-sided t test ( n > 4). ** p

    Article Snippet: The following primary antibodies were used in this study: anti-βIII-tubulin (rabbit IgG; Covance, PRB-435P; 1:500), anti-glial fibrillary acidic protein (GFAP; rabbit IgG; DAKO, Z0334; 1:500), anti-GFAP (mouse IgG; Sigma-Aldrich, G3893; 1:200), anti-fatty acid binding protein 7 (FABP7; rabbit IgG; Millipore, AB9558; 1:250), anti-Sox21 (goat IgG; R & D Systems, AF3538; 1:100), anti-Sox2 (rabbit IgG; Millipore, AB5603; 1:200), anti-Sox1/(2)/3 (rabbit IgG; gift from Dr. H. Kondoh, Osaka University, Osaka, Japan; 1:5000) , anti-Nestin (mouse IgG; BD Biosciences/PharMingen, 556309; 1:200), anti-doublecortin (DCX; goat IgG; Santa Cruz Biotechnology, sc-8066; 1:100), anti- DCX (rabbit IgG; Abcam, ab18723; 1:100), anti-polysialylated neuronal cell adhesion molecule (PSA-NCAM; mouse IgMκ; gift from Dr. T. Seki, Tokyo Medical University, Tokyo, Japan; 1:200), anti-NeuN (mouse IgG; Millipore, MAB377; 1:100), anti-S100β (mouse IgG; Sigma-Aldrich, S2532; 1:200), anti-glutathione S -transferase π (mouse IgG; BD Biosciences/PharMingen, 610719; 1:200), anti-Brn2 (Santa Cruz Biotechnology, sc-6029; 1:100), anti-Tbr1 (rabbit IgG; gift from Dr. R. F. Hevner, University of Washington, Seattle, WA; 1:10,000), anti-Ascl1 (mouse IgG; BD Biosciences/PharMingen, 556604; 1:100), anti-Tbr2 (rabbit IgG; Abcam, ab23345; 1:200), anti-Ki67 (rabbit IgG; Abcam, ab15580; 1:3000), anti-GFP (rabbit IgG, Medical and Biological Laboratories, 598, 1:500; goat IgG, Rockland, 600-101-215, 1:500), anti-BrdU (rat IgG; Abcam, ab6326; 1:100), anti-lacZ (rabbit IgG, Cappel, 55976, 1:500; goat IgG, Cappel, 56028, 1:500), and normal IgG (rabbit IgG, Millipore, DAM1421465; goat IgG, Santa Cruz Biotechnology, sc-2028).

    Techniques: Activity Assay, Transfection, Luciferase, Construct, Expressing, Plasmid Preparation, Binding Assay, Sequencing