mouse anti gfap  (Thermo Fisher)


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    Structured Review

    Thermo Fisher mouse anti gfap
    <t>NALP1</t> and <t>GFAP</t> double staining in the spinal dorsal horn of mice chronically treated with DMSO or curcumin (120 mg/kg) on the day 7 after sham or SNI surgery. The sham surgery induced a weak NALP1-IR in the ipsilateral spinal dorsal horn ( A,B ). Changes in the NALP1-IR were not observed in the sham group after intraperitoneal curcumin administration ( A,B ). After SNI surgery, increased NALP1-IR and GFAP-IR were detected ipsilateral to the lesion of the spinal dorsal horn, and a large part of the NALP1-IRs were co-labelled with GFAP-IR, indicating that NALP1 accumulated in the spinal cord astrocytes on the day 7 after SNI ( C,C ’). The intraperitoneal administration of curcumin inhibited the expression of NALP1-IR in the ipsilateral spinal dorsal horn after SNI ( D ). ( E ) Intensity of NALP1 staining in the superficial dorsal horn (laminae I–III). *p
    Mouse Anti Gfap, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Curcumin ameliorates neuropathic pain by down-regulating spinal IL-1β via suppressing astroglial NALP1 inflammasome and JAK2-STAT3 signalling"

    Article Title: Curcumin ameliorates neuropathic pain by down-regulating spinal IL-1β via suppressing astroglial NALP1 inflammasome and JAK2-STAT3 signalling

    Journal: Scientific Reports

    doi: 10.1038/srep28956

    NALP1 and GFAP double staining in the spinal dorsal horn of mice chronically treated with DMSO or curcumin (120 mg/kg) on the day 7 after sham or SNI surgery. The sham surgery induced a weak NALP1-IR in the ipsilateral spinal dorsal horn ( A,B ). Changes in the NALP1-IR were not observed in the sham group after intraperitoneal curcumin administration ( A,B ). After SNI surgery, increased NALP1-IR and GFAP-IR were detected ipsilateral to the lesion of the spinal dorsal horn, and a large part of the NALP1-IRs were co-labelled with GFAP-IR, indicating that NALP1 accumulated in the spinal cord astrocytes on the day 7 after SNI ( C,C ’). The intraperitoneal administration of curcumin inhibited the expression of NALP1-IR in the ipsilateral spinal dorsal horn after SNI ( D ). ( E ) Intensity of NALP1 staining in the superficial dorsal horn (laminae I–III). *p
    Figure Legend Snippet: NALP1 and GFAP double staining in the spinal dorsal horn of mice chronically treated with DMSO or curcumin (120 mg/kg) on the day 7 after sham or SNI surgery. The sham surgery induced a weak NALP1-IR in the ipsilateral spinal dorsal horn ( A,B ). Changes in the NALP1-IR were not observed in the sham group after intraperitoneal curcumin administration ( A,B ). After SNI surgery, increased NALP1-IR and GFAP-IR were detected ipsilateral to the lesion of the spinal dorsal horn, and a large part of the NALP1-IRs were co-labelled with GFAP-IR, indicating that NALP1 accumulated in the spinal cord astrocytes on the day 7 after SNI ( C,C ’). The intraperitoneal administration of curcumin inhibited the expression of NALP1-IR in the ipsilateral spinal dorsal horn after SNI ( D ). ( E ) Intensity of NALP1 staining in the superficial dorsal horn (laminae I–III). *p

    Techniques Used: Double Staining, Mouse Assay, Expressing, Staining

    2) Product Images from "Elevated p62/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide"

    Article Title: Elevated p62/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201507023

    p62 KO rescues the maintenance and differentiation defects of Fip200-null NSCs in vivo. (A) H E staining of SVZ and hippocampus from FIP GFAP cKO and 2cKO mice at P28. Dotted lines indicate boundaries of SVZ and DG ( n = 4 mice each). (B) Mean ± SEM of SVZ cellularity (left) and DG area (right) per section from Ctrl, FIP GFAP cKO, 2cKO, and p62 KO mice at P28. (C and D) Immunofluorescence for GFAP, Nestin (C), SOX2 (D), and DAPI in SVZ from Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO mice at P28. Mean ± SEM of GFAP+ and Nestin+ (C) and GFAP+ and Sox2+ (D) cell number per SVZ section ( n = 4 mice each). (E) Immunofluorescence of long-term retained BrdU and DAPI in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of BrdU+ cell number per SVZ section ( n = 4 mice each). (F and G) Immunofluorescence for p62 and DAPI (F; n = 3 mice each) and TUNEL and DAPI (G) in SVZ of Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO mice at P28. Mean ± SEM of TUNEL+ cells per 100 SVZ cells (G, n = 4 mice each, > 1,000 cells per mouse). (H and I) Immunofluorescence of Nestin, GFAP, and short-term labeled BrdU in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of the percentage of GFAP + Nestin + BrdU + to total GFAP + Nestin + cells (H) or total BrdU + cells (I) per section ( n = 5 mice each). (J and K) Immunofluorescence for Nestin, GFAP, and Ki67 in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of the percentage of GFAP + Nestin + Ki67 + to total GFAP + Nestin + cells (J) or total Ki67 + cells (K) per section ( n = 5 mice each). (L and M) Immunofluorescence for DCX in SVZ (L) and NeuN in olfactory bulb (M) in Ctrl, Fip200 GFAP cKO, 2cKO and p62 KO mice at P28. Mean ± SEM of DCX + cells per section (L) and NeuN + cells per square millimeter (M; n = 4 mice each). Dotted lines indicate the boundaries of SVZ (C–G). CC, corpus callosum; E, ependymal layer; LV, lateral ventricle; OB, olfactory bulb; RMS, rostral migratory stream; ST, striatum. Bars: (A, E, and G and insets) 50 µm; (C, D, and insets) 25 µm; (F and insets) 15 µm. *, P
    Figure Legend Snippet: p62 KO rescues the maintenance and differentiation defects of Fip200-null NSCs in vivo. (A) H E staining of SVZ and hippocampus from FIP GFAP cKO and 2cKO mice at P28. Dotted lines indicate boundaries of SVZ and DG ( n = 4 mice each). (B) Mean ± SEM of SVZ cellularity (left) and DG area (right) per section from Ctrl, FIP GFAP cKO, 2cKO, and p62 KO mice at P28. (C and D) Immunofluorescence for GFAP, Nestin (C), SOX2 (D), and DAPI in SVZ from Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO mice at P28. Mean ± SEM of GFAP+ and Nestin+ (C) and GFAP+ and Sox2+ (D) cell number per SVZ section ( n = 4 mice each). (E) Immunofluorescence of long-term retained BrdU and DAPI in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of BrdU+ cell number per SVZ section ( n = 4 mice each). (F and G) Immunofluorescence for p62 and DAPI (F; n = 3 mice each) and TUNEL and DAPI (G) in SVZ of Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO mice at P28. Mean ± SEM of TUNEL+ cells per 100 SVZ cells (G, n = 4 mice each, > 1,000 cells per mouse). (H and I) Immunofluorescence of Nestin, GFAP, and short-term labeled BrdU in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of the percentage of GFAP + Nestin + BrdU + to total GFAP + Nestin + cells (H) or total BrdU + cells (I) per section ( n = 5 mice each). (J and K) Immunofluorescence for Nestin, GFAP, and Ki67 in Ctrl, Fip200 GFAP cKO, 2cKO, and p62 KO SVZ at P28. Mean ± SEM of the percentage of GFAP + Nestin + Ki67 + to total GFAP + Nestin + cells (J) or total Ki67 + cells (K) per section ( n = 5 mice each). (L and M) Immunofluorescence for DCX in SVZ (L) and NeuN in olfactory bulb (M) in Ctrl, Fip200 GFAP cKO, 2cKO and p62 KO mice at P28. Mean ± SEM of DCX + cells per section (L) and NeuN + cells per square millimeter (M; n = 4 mice each). Dotted lines indicate the boundaries of SVZ (C–G). CC, corpus callosum; E, ependymal layer; LV, lateral ventricle; OB, olfactory bulb; RMS, rostral migratory stream; ST, striatum. Bars: (A, E, and G and insets) 50 µm; (C, D, and insets) 25 µm; (F and insets) 15 µm. *, P

    Techniques Used: In Vivo, Staining, Mouse Assay, Immunofluorescence, TUNEL Assay, Labeling

    3) Product Images from "Interferon-stimulated gene 15 as a general marker for acute and chronic neuronal injuries"

    Article Title: Interferon-stimulated gene 15 as a general marker for acute and chronic neuronal injuries

    Journal: Sheng li xue bao : [Acta physiologica Sinica]

    doi:

    Both common carotid arteries (BCCA) occlusion-induced increase of ISG15 expression in the hippocampus. Immunohistochemistry showing strong expression of ISG15 ( A ), GFAP ( B ), and Iba-1 ( C ) in the hippocampus of mice subjected to BCCA occlusion but not sham controls ( D , E , F ). Double-staining showing the co-localization of ISG15 ( G ) with GFAP-positive cells ( H ) in the hippocampus. ( I ): overlap of ( G ) and ( H ).
    Figure Legend Snippet: Both common carotid arteries (BCCA) occlusion-induced increase of ISG15 expression in the hippocampus. Immunohistochemistry showing strong expression of ISG15 ( A ), GFAP ( B ), and Iba-1 ( C ) in the hippocampus of mice subjected to BCCA occlusion but not sham controls ( D , E , F ). Double-staining showing the co-localization of ISG15 ( G ) with GFAP-positive cells ( H ) in the hippocampus. ( I ): overlap of ( G ) and ( H ).

    Techniques Used: Expressing, Immunohistochemistry, Mouse Assay, Double Staining

    4) Product Images from "In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses"

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses

    Journal: Journal of Virology

    doi: 10.1128/JVI.77.18.9799-9808.2003

    GFP expression in mouse brains after cerebral injections of AcVSVG-CAGFP. Mice were injected with 4 × 10 7 PFU of AcVSVG-CAGFP in the right lateral ventricle. GFP expression in the brain was examined by fluorescent stereomicroscopy 2 days after injection. (A) Panels A to D are stereomicroscopic images of whole brain (A and B) and brain cross sections (C and D). Panels A and C are bright-field views, while panels B and D are fluorescent views. Arrows and dark staining indicate the injection route, as the infiltrated viral inoculum contained 0.04% trypan blue. (B) Immunohistochemical staining of the cryostat sections was examined by fluorescence microscopy following staining with antibodies specific for GFP (A and D), GFAP as a glial marker (B), or MAP2 as a neuronal marker (E). Panels C and F are merged images.
    Figure Legend Snippet: GFP expression in mouse brains after cerebral injections of AcVSVG-CAGFP. Mice were injected with 4 × 10 7 PFU of AcVSVG-CAGFP in the right lateral ventricle. GFP expression in the brain was examined by fluorescent stereomicroscopy 2 days after injection. (A) Panels A to D are stereomicroscopic images of whole brain (A and B) and brain cross sections (C and D). Panels A and C are bright-field views, while panels B and D are fluorescent views. Arrows and dark staining indicate the injection route, as the infiltrated viral inoculum contained 0.04% trypan blue. (B) Immunohistochemical staining of the cryostat sections was examined by fluorescence microscopy following staining with antibodies specific for GFP (A and D), GFAP as a glial marker (B), or MAP2 as a neuronal marker (E). Panels C and F are merged images.

    Techniques Used: Expressing, Mouse Assay, Injection, Staining, Immunohistochemistry, Fluorescence, Microscopy, Marker

    Gene transduction into primary rat cerebellar cells with AcVSVG-CAGFP. Primary rat cerebellar and hippocampal cultures were infected with AcVSVG-CAGFP. Immunofluorescence was examined by confocal microscopy. (A, D, and G) Anti-GFP immunochemistry. (B) Anti-Calbindin immunochemistry as a purkinje marker. (E) Anti-GFAP immunochemistry as a glial marker. (H) Anti-MAP2 immunochemistry as a neuronal marker. C, F, and I are merged images.
    Figure Legend Snippet: Gene transduction into primary rat cerebellar cells with AcVSVG-CAGFP. Primary rat cerebellar and hippocampal cultures were infected with AcVSVG-CAGFP. Immunofluorescence was examined by confocal microscopy. (A, D, and G) Anti-GFP immunochemistry. (B) Anti-Calbindin immunochemistry as a purkinje marker. (E) Anti-GFAP immunochemistry as a glial marker. (H) Anti-MAP2 immunochemistry as a neuronal marker. C, F, and I are merged images.

    Techniques Used: Transduction, Infection, Immunofluorescence, Confocal Microscopy, Marker

    5) Product Images from "Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells"

    Article Title: Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5087-12.2013

    HFS activates microglia and astrocytes. Representative experiments show the changes in microglial ( A–C ) and astrocytic ( D–F ) specific markers after HFS of primary afferent fibers. A , D , Immunoreactivity for the microglia-specific marker Iba1 ( A , n = 15 slices) and the astrocyte-specific marker GFAP ( D , n = 16 slices) in untreated slices. B , 30 min after HFS, Iba1 immunofluorescence is increased on the ipsilateral site (right) but not contralaterally to HFS ( n = 14 slices). C , 120 min after HFS, Iba1 levels are back to control again ( n = 14 slices). E , Thirty minutes after HFS, GFAP immunoreactivity is unchanged compared with control ( n = 16 slices). F , Two hours after HFS of primary afferents, GFAP shows increased immunoreactivity on the ipsilateral (right) and contralateral (left) side ( n = 16 slices). G , Quantification of Iba1 and GFAP intensity 30 and 120 min after HFS. * p
    Figure Legend Snippet: HFS activates microglia and astrocytes. Representative experiments show the changes in microglial ( A–C ) and astrocytic ( D–F ) specific markers after HFS of primary afferent fibers. A , D , Immunoreactivity for the microglia-specific marker Iba1 ( A , n = 15 slices) and the astrocyte-specific marker GFAP ( D , n = 16 slices) in untreated slices. B , 30 min after HFS, Iba1 immunofluorescence is increased on the ipsilateral site (right) but not contralaterally to HFS ( n = 14 slices). C , 120 min after HFS, Iba1 levels are back to control again ( n = 14 slices). E , Thirty minutes after HFS, GFAP immunoreactivity is unchanged compared with control ( n = 16 slices). F , Two hours after HFS of primary afferents, GFAP shows increased immunoreactivity on the ipsilateral (right) and contralateral (left) side ( n = 16 slices). G , Quantification of Iba1 and GFAP intensity 30 and 120 min after HFS. * p

    Techniques Used: Marker, Immunofluorescence

    Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.
    Figure Legend Snippet: Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.

    Techniques Used: Double Immunostaining, Marker

    6) Product Images from "PlexinA4 Distribution in the Adult Rat Spinal Cord and Dorsal Root Ganglia"

    Article Title: PlexinA4 Distribution in the Adult Rat Spinal Cord and Dorsal Root Ganglia

    Journal: Journal of Chemical Neuroanatomy

    doi: 10.1016/j.jchemneu.2012.03.002

    PlexinA4 in adult spinal cord A) Aliquots of rat and mouse spinal cord tissue were subjected to SDS/7.5% PAGE and immunoblotted with polyclonal anti-PlexinA4 antibodies. Molecular mass standards (in kDa) are indicated. A band of approximately 210 kDa is detected in both rat and mouse tissue (arrow). Immunoreactivity is abolished when antibodies are preabsorbed with corresponding peptide and when primary antibodies are omitted. Micrographs of HEK293 cells transfected with PlexinA4 or PlexinA1 expressing plasmids co-stained with myc-tag and PlexinA4 antibodies. The PlexinA4 antibodies only immunostained cells expressing PlexinA4. B) Coronal and sagittal sections from adult rat spinal cord immunolabelled with PlexinA4 antibodies. PlexinA4 immunostaining is visible at every spinal cord level in both the dorsal and ventral horns, as well as white matter tracts. PlexinA4 staining is visible in neurons and fibers in the dorsal and ventral horns. At each level, the placement of the sagittal section is indicated on the coronal sections by a dashed line. C) Representative micrographs from PlexinA4 (green) and NeuN (red) or GFAP (red) co-immunostained rat cervical ventral horn as indicated. All PlexinA4 positive cells are also NeuN positive suggesting that PlexinA4 is expressed by neurons (arrowheads). There is no overlap between PlexinA4 and GFAP staining suggesting that PlexinA4 is expressed by neurons (arrowheads) but not by astroglia (arrows). Abbreviations: D: dorsal, V: ventral. Scale bars: A: 10μm, B: 0.5 mm, C: 50 μm.
    Figure Legend Snippet: PlexinA4 in adult spinal cord A) Aliquots of rat and mouse spinal cord tissue were subjected to SDS/7.5% PAGE and immunoblotted with polyclonal anti-PlexinA4 antibodies. Molecular mass standards (in kDa) are indicated. A band of approximately 210 kDa is detected in both rat and mouse tissue (arrow). Immunoreactivity is abolished when antibodies are preabsorbed with corresponding peptide and when primary antibodies are omitted. Micrographs of HEK293 cells transfected with PlexinA4 or PlexinA1 expressing plasmids co-stained with myc-tag and PlexinA4 antibodies. The PlexinA4 antibodies only immunostained cells expressing PlexinA4. B) Coronal and sagittal sections from adult rat spinal cord immunolabelled with PlexinA4 antibodies. PlexinA4 immunostaining is visible at every spinal cord level in both the dorsal and ventral horns, as well as white matter tracts. PlexinA4 staining is visible in neurons and fibers in the dorsal and ventral horns. At each level, the placement of the sagittal section is indicated on the coronal sections by a dashed line. C) Representative micrographs from PlexinA4 (green) and NeuN (red) or GFAP (red) co-immunostained rat cervical ventral horn as indicated. All PlexinA4 positive cells are also NeuN positive suggesting that PlexinA4 is expressed by neurons (arrowheads). There is no overlap between PlexinA4 and GFAP staining suggesting that PlexinA4 is expressed by neurons (arrowheads) but not by astroglia (arrows). Abbreviations: D: dorsal, V: ventral. Scale bars: A: 10μm, B: 0.5 mm, C: 50 μm.

    Techniques Used: Polyacrylamide Gel Electrophoresis, Transfection, Expressing, Staining, Immunostaining

    7) Product Images from "Fgf-Dependent Glial Cell Bridges Facilitate Spinal Cord Regeneration in Zebrafish"

    Article Title: Fgf-Dependent Glial Cell Bridges Facilitate Spinal Cord Regeneration in Zebrafish

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0758-12.2012

    Fgf ligands and their downstream target genes are induced in glia and neurons after SCI. a–o , Sections from Tg( GFAP:GFP ) transgenic fish documenting the location of GFAP-positive glia or traced neurons on each section before and after in situ hybridization. a–e , Intact control spinal cords. fgf8a is expressed at low levels on GFAP-positive glia cells in the ventricular zone ( a‴ and a⁗ show boxed region in a , a′ ) and fgf3 is expressed at low levels on large neurons ( b , arrowhead). spry4 is expressed at low levels in neurons ( c , arrowhead), whereas pea3 ( d ) and erm ( e ) are almost undetectable in adjacent sections. f–j , Three days after SCI. fgf8a expression is significantly increased on GFAP-positive glia cells found at the lesion site ( f′ is a higher magnification of boxed area in f and f‴ and f⁗ are higher magnifications of boxed region in f′ ). fgf3 expression increases in both neurons ( g ; g′ , boxed area in g , arrowhead) and in glia (arrow) at the cc surrounding the lesion site. spry4 expression is high in neurons ( h , arrowheads) and glia (arrows), while pea3 ( i ) and erm ( j ) expression is upregulated on glia cells. ( k–o , 2–3 weeks after SCI. fgf8a is highly expressed ( k ; k′ , boxed area in k ) in glia cells at the lesion site and around the cc. fgf3 expression is decreased in glia cells at the lesion site (l, arrow), but increased in neurons upstream of the lesion (l ′ , arrowhead). ( l″ ) fgf3 -expressing cells are identified as neurons by colabeling with NeuN and by accumulation of TMRD axonal tracer labeling ( n , arrowheads). spry4 expression is increased in neurons ( m , arrowheads), which are also labeled by TMRD tracer ( o , arrowheads). p , q , FgfR2 is upregulated on GFAP-positive glia cells (arrowheads) and neurons (arrows) 2 weeks post-SCI in injured ( p ; p′–p‴ , boxed area in p ) compared with intact ( q ; q′–q‴ , boxed area in q ) spinal cords. Caudal side of spinal cord to the left in all panels. cc, Central canal. Representative results from at least n = 4 fish in each condition. Scale bars: a , a′ , f , f′ , f″ , g , k , 100 μm; in high-power magnification panels, including a‴ , b–e , f‴ , g′–j , k′–o′ , p , q , 50 μm; p′–p‴ , q′-q‴ , 10 μm.
    Figure Legend Snippet: Fgf ligands and their downstream target genes are induced in glia and neurons after SCI. a–o , Sections from Tg( GFAP:GFP ) transgenic fish documenting the location of GFAP-positive glia or traced neurons on each section before and after in situ hybridization. a–e , Intact control spinal cords. fgf8a is expressed at low levels on GFAP-positive glia cells in the ventricular zone ( a‴ and a⁗ show boxed region in a , a′ ) and fgf3 is expressed at low levels on large neurons ( b , arrowhead). spry4 is expressed at low levels in neurons ( c , arrowhead), whereas pea3 ( d ) and erm ( e ) are almost undetectable in adjacent sections. f–j , Three days after SCI. fgf8a expression is significantly increased on GFAP-positive glia cells found at the lesion site ( f′ is a higher magnification of boxed area in f and f‴ and f⁗ are higher magnifications of boxed region in f′ ). fgf3 expression increases in both neurons ( g ; g′ , boxed area in g , arrowhead) and in glia (arrow) at the cc surrounding the lesion site. spry4 expression is high in neurons ( h , arrowheads) and glia (arrows), while pea3 ( i ) and erm ( j ) expression is upregulated on glia cells. ( k–o , 2–3 weeks after SCI. fgf8a is highly expressed ( k ; k′ , boxed area in k ) in glia cells at the lesion site and around the cc. fgf3 expression is decreased in glia cells at the lesion site (l, arrow), but increased in neurons upstream of the lesion (l ′ , arrowhead). ( l″ ) fgf3 -expressing cells are identified as neurons by colabeling with NeuN and by accumulation of TMRD axonal tracer labeling ( n , arrowheads). spry4 expression is increased in neurons ( m , arrowheads), which are also labeled by TMRD tracer ( o , arrowheads). p , q , FgfR2 is upregulated on GFAP-positive glia cells (arrowheads) and neurons (arrows) 2 weeks post-SCI in injured ( p ; p′–p‴ , boxed area in p ) compared with intact ( q ; q′–q‴ , boxed area in q ) spinal cords. Caudal side of spinal cord to the left in all panels. cc, Central canal. Representative results from at least n = 4 fish in each condition. Scale bars: a , a′ , f , f′ , f″ , g , k , 100 μm; in high-power magnification panels, including a‴ , b–e , f‴ , g′–j , k′–o′ , p , q , 50 μm; p′–p‴ , q′-q‴ , 10 μm.

    Techniques Used: Transgenic Assay, Fluorescence In Situ Hybridization, In Situ Hybridization, Expressing, Labeling

    8) Product Images from "Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells"

    Article Title: Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5087-12.2013

    Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.
    Figure Legend Snippet: Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.

    Techniques Used: Double Immunostaining, Marker

    9) Product Images from "Fgf-Dependent Glial Cell Bridges Facilitate Spinal Cord Regeneration in Zebrafish"

    Article Title: Fgf-Dependent Glial Cell Bridges Facilitate Spinal Cord Regeneration in Zebrafish

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0758-12.2012

    Spry4 expression after spinal cord injury in mouse. a , RT-PCR of RNA extracted from the brain, liver, muscle and spinal cord of two uninjured mice shows spry4 expression primarily in the brain and spinal cord, with some expression detected in the liver. GAPDH was used as a PCR amplification control. b , b′ , Spry4 is expressed in GFAP-expressing astrocytes in the gray matter of the intact spinal cord (arrowheads). c–c″ , Four days post-SCI. Spry4 is upregulated at the lesion site on GFAP-positive reactive astrocytes. d , d′ , High-power magnification of an astrocyte at the lesion site, which expresses high levels of GFAP and Spry4. Scale bars: b , d , 25 μm; c , 100 μm.
    Figure Legend Snippet: Spry4 expression after spinal cord injury in mouse. a , RT-PCR of RNA extracted from the brain, liver, muscle and spinal cord of two uninjured mice shows spry4 expression primarily in the brain and spinal cord, with some expression detected in the liver. GAPDH was used as a PCR amplification control. b , b′ , Spry4 is expressed in GFAP-expressing astrocytes in the gray matter of the intact spinal cord (arrowheads). c–c″ , Four days post-SCI. Spry4 is upregulated at the lesion site on GFAP-positive reactive astrocytes. d , d′ , High-power magnification of an astrocyte at the lesion site, which expresses high levels of GFAP and Spry4. Scale bars: b , d , 25 μm; c , 100 μm.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Polymerase Chain Reaction, Amplification

    Fgf ligands and their downstream target genes are induced in glia and neurons after SCI. a–o , Sections from Tg( GFAP:GFP ) transgenic fish documenting the location of GFAP-positive glia or traced neurons on each section before and after in situ hybridization. a–e , Intact control spinal cords. fgf8a is expressed at low levels on GFAP-positive glia cells in the ventricular zone ( a‴ and a⁗ show boxed region in a , a′ ) and fgf3 is expressed at low levels on large neurons ( b , arrowhead). spry4 is expressed at low levels in neurons ( c , arrowhead), whereas pea3 ( d ) and erm ( e ) are almost undetectable in adjacent sections. f–j , Three days after SCI. fgf8a expression is significantly increased on GFAP-positive glia cells found at the lesion site ( f′ is a higher magnification of boxed area in f and f‴ and f⁗ are higher magnifications of boxed region in f′ ). fgf3 expression increases in both neurons ( g ; g′ , boxed area in g , arrowhead) and in glia (arrow) at the cc surrounding the lesion site. spry4 expression is high in neurons ( h , arrowheads) and glia (arrows), while pea3 ( i ) and erm ( j ) expression is upregulated on glia cells. ( k–o , 2–3 weeks after SCI. fgf8a is highly expressed ( k ; k′ , boxed area in k ) in glia cells at the lesion site and around the cc. fgf3 expression is decreased in glia cells at the lesion site (l, arrow), but increased in neurons upstream of the lesion (l ′ , arrowhead). ( l″ ) fgf3 -expressing cells are identified as neurons by colabeling with NeuN and by accumulation of TMRD axonal tracer labeling ( n , arrowheads). spry4 expression is increased in neurons ( m , arrowheads), which are also labeled by TMRD tracer ( o , arrowheads). p , q , FgfR2 is upregulated on GFAP-positive glia cells (arrowheads) and neurons (arrows) 2 weeks post-SCI in injured ( p ; p′–p‴ , boxed area in p ) compared with intact ( q ; q′–q‴ , boxed area in q ) spinal cords. Caudal side of spinal cord to the left in all panels. cc, Central canal. Representative results from at least n = 4 fish in each condition. Scale bars: a , a′ , f , f′ , f″ , g , k , 100 μm; in high-power magnification panels, including a‴ , b–e , f‴ , g′–j , k′–o′ , p , q , 50 μm; p′–p‴ , q′-q‴ , 10 μm.
    Figure Legend Snippet: Fgf ligands and their downstream target genes are induced in glia and neurons after SCI. a–o , Sections from Tg( GFAP:GFP ) transgenic fish documenting the location of GFAP-positive glia or traced neurons on each section before and after in situ hybridization. a–e , Intact control spinal cords. fgf8a is expressed at low levels on GFAP-positive glia cells in the ventricular zone ( a‴ and a⁗ show boxed region in a , a′ ) and fgf3 is expressed at low levels on large neurons ( b , arrowhead). spry4 is expressed at low levels in neurons ( c , arrowhead), whereas pea3 ( d ) and erm ( e ) are almost undetectable in adjacent sections. f–j , Three days after SCI. fgf8a expression is significantly increased on GFAP-positive glia cells found at the lesion site ( f′ is a higher magnification of boxed area in f and f‴ and f⁗ are higher magnifications of boxed region in f′ ). fgf3 expression increases in both neurons ( g ; g′ , boxed area in g , arrowhead) and in glia (arrow) at the cc surrounding the lesion site. spry4 expression is high in neurons ( h , arrowheads) and glia (arrows), while pea3 ( i ) and erm ( j ) expression is upregulated on glia cells. ( k–o , 2–3 weeks after SCI. fgf8a is highly expressed ( k ; k′ , boxed area in k ) in glia cells at the lesion site and around the cc. fgf3 expression is decreased in glia cells at the lesion site (l, arrow), but increased in neurons upstream of the lesion (l ′ , arrowhead). ( l″ ) fgf3 -expressing cells are identified as neurons by colabeling with NeuN and by accumulation of TMRD axonal tracer labeling ( n , arrowheads). spry4 expression is increased in neurons ( m , arrowheads), which are also labeled by TMRD tracer ( o , arrowheads). p , q , FgfR2 is upregulated on GFAP-positive glia cells (arrowheads) and neurons (arrows) 2 weeks post-SCI in injured ( p ; p′–p‴ , boxed area in p ) compared with intact ( q ; q′–q‴ , boxed area in q ) spinal cords. Caudal side of spinal cord to the left in all panels. cc, Central canal. Representative results from at least n = 4 fish in each condition. Scale bars: a , a′ , f , f′ , f″ , g , k , 100 μm; in high-power magnification panels, including a‴ , b–e , f‴ , g′–j , k′–o′ , p , q , 50 μm; p′–p‴ , q′-q‴ , 10 μm.

    Techniques Used: Transgenic Assay, Fluorescence In Situ Hybridization, In Situ Hybridization, Expressing, Labeling

    Fgf-dependent MAPK signaling is induced in glia and neurons after spinal cord injury. a , b , In intact control spinal cords p-MAPK levels are low in GFAP-expressing glia along the central canal (cc) ( a–a″ , arrowheads), but become strongly upregulated 24 h post-SCI ( b ) ( n = 5). c–e , Nestin levels at the cc are also low ( n = 4) in intact spinal cord, but both p-MAPK and nestin levels increase shortly after SCI (24 h, d ; 3 d, e , arrowheads) in glia cells at the cc ( n = 5). f–l , MAPK is activated by Fgf signaling after SCI as confirmed by Western blot analysis of protein derived from spinal cord lesion sites in wild-type, spry4 −/− and dn-fgfr1 lines after SCI ( k ). In WT ( f ) and DMSO only control ( g ), p-MAPK levels increase 24 h post-SCI ( f′ , g′ , k ). When Fgf is inhibited by SU5402 injection ( h ) or heat shock of hsp70l:dn-fgfr1-EGFP fish ( i ), this increase in p-MAPK is abolished ( h′ , i′ , k ). Conversely, in spry4 −/− fish p-MAPK activity is increased in GFAP-positive cells as in wild-type, and additionally in neurons ( j″ , arrowheads) that colabel with the neural NeuN marker ( l–l″ ). Scale bars: a–j , 50 μm; l , 10 μm.
    Figure Legend Snippet: Fgf-dependent MAPK signaling is induced in glia and neurons after spinal cord injury. a , b , In intact control spinal cords p-MAPK levels are low in GFAP-expressing glia along the central canal (cc) ( a–a″ , arrowheads), but become strongly upregulated 24 h post-SCI ( b ) ( n = 5). c–e , Nestin levels at the cc are also low ( n = 4) in intact spinal cord, but both p-MAPK and nestin levels increase shortly after SCI (24 h, d ; 3 d, e , arrowheads) in glia cells at the cc ( n = 5). f–l , MAPK is activated by Fgf signaling after SCI as confirmed by Western blot analysis of protein derived from spinal cord lesion sites in wild-type, spry4 −/− and dn-fgfr1 lines after SCI ( k ). In WT ( f ) and DMSO only control ( g ), p-MAPK levels increase 24 h post-SCI ( f′ , g′ , k ). When Fgf is inhibited by SU5402 injection ( h ) or heat shock of hsp70l:dn-fgfr1-EGFP fish ( i ), this increase in p-MAPK is abolished ( h′ , i′ , k ). Conversely, in spry4 −/− fish p-MAPK activity is increased in GFAP-positive cells as in wild-type, and additionally in neurons ( j″ , arrowheads) that colabel with the neural NeuN marker ( l–l″ ). Scale bars: a–j , 50 μm; l , 10 μm.

    Techniques Used: Expressing, Western Blot, Derivative Assay, Injection, Fluorescence In Situ Hybridization, Activity Assay, Marker

    Fgf signaling regulates glia cell proliferation and migration. a–e , At 3 weeks post-SCI, Fgf inhibition by SU5402 injection ( n = 6) ( c ) or heat shock of hsp70l:dn-fgfr1-EGFP ( n = 7) ( d ) causes a significant reduction ( f ) in proliferation (assayed by BrdU incorporation) compared with WT DMSO ( a ) or wt heat shock ( b ) fish. In contrast, the delayed heat shock of hsp70l:dn-fgfr1-EGFP (initiated only at 6 d post-SCI) does not show a reduction in proliferation ( e , g ). h–n , Similarly, the resulting migration and accumulation of cells (DAPI labeled) at the lesion site is also reduced in SU5402-injected or heat-shocked hsp70l:dn-fgfr1-EGFP ( h ) fish compared with WT DMSO ( n = 6) ( j ) or heat-shocked fish ( n = 6) ( k ). Again delaying the heat shock by 5 d completely ameliorates the effect and there is no significant change in the number of cells that migrate and accumulate at the lesion in this treatment ( n = 6) ( h , n ). i , Assessing the degree of proliferation across different time periods reveals that the bulk of the proliferative response occurs within the first 5 d post-SCI ( n = 3 per time point). o–q , In contrast, Fgf signaling upregulation in spry4 −/− mutants ( o , n = 7) or Fgf8-injected fish ( p , n = 6) results in a significant increase in proliferation already by 5 d post-SCI ( q ). Additionally, glia show an accelerated differentiation and earlier expression of GFAP:GFP in both of these increased Fgf signaling conditions ( o–q ). r–t , Differentiation of the bipolar glial morphology is correlated with MAPK signaling activity in wild-type and hsp70:dn-fgfr1 fish with delayed heat shock starting 6 d after SCI. Scale bars: a–e , j–n–p , 100 μm; o′ , r, s′ , 50 μm. Results are presented in f–i , q , and t as mean ± SEM, ** p
    Figure Legend Snippet: Fgf signaling regulates glia cell proliferation and migration. a–e , At 3 weeks post-SCI, Fgf inhibition by SU5402 injection ( n = 6) ( c ) or heat shock of hsp70l:dn-fgfr1-EGFP ( n = 7) ( d ) causes a significant reduction ( f ) in proliferation (assayed by BrdU incorporation) compared with WT DMSO ( a ) or wt heat shock ( b ) fish. In contrast, the delayed heat shock of hsp70l:dn-fgfr1-EGFP (initiated only at 6 d post-SCI) does not show a reduction in proliferation ( e , g ). h–n , Similarly, the resulting migration and accumulation of cells (DAPI labeled) at the lesion site is also reduced in SU5402-injected or heat-shocked hsp70l:dn-fgfr1-EGFP ( h ) fish compared with WT DMSO ( n = 6) ( j ) or heat-shocked fish ( n = 6) ( k ). Again delaying the heat shock by 5 d completely ameliorates the effect and there is no significant change in the number of cells that migrate and accumulate at the lesion in this treatment ( n = 6) ( h , n ). i , Assessing the degree of proliferation across different time periods reveals that the bulk of the proliferative response occurs within the first 5 d post-SCI ( n = 3 per time point). o–q , In contrast, Fgf signaling upregulation in spry4 −/− mutants ( o , n = 7) or Fgf8-injected fish ( p , n = 6) results in a significant increase in proliferation already by 5 d post-SCI ( q ). Additionally, glia show an accelerated differentiation and earlier expression of GFAP:GFP in both of these increased Fgf signaling conditions ( o–q ). r–t , Differentiation of the bipolar glial morphology is correlated with MAPK signaling activity in wild-type and hsp70:dn-fgfr1 fish with delayed heat shock starting 6 d after SCI. Scale bars: a–e , j–n–p , 100 μm; o′ , r, s′ , 50 μm. Results are presented in f–i , q , and t as mean ± SEM, ** p

    Techniques Used: Migration, Inhibition, Injection, BrdU Incorporation Assay, Fluorescence In Situ Hybridization, Labeling, Expressing, Activity Assay

    10) Product Images from "Differentiated Adipose-derived Stem Cells Promote Reinnervation of Rat Skin Flaps"

    Article Title: Differentiated Adipose-derived Stem Cells Promote Reinnervation of Rat Skin Flaps

    Journal: Plastic and Reconstructive Surgery Global Open

    doi: 10.1097/GOX.0b013e318299134d

    A, Fibroblast-like ASCs (left) changed to bipolar, spindle-shaped dASCs (middle), which were similar to SCs (right). Scale bar: 50 µm. B, Western blotting analysis of cell lysates showed that ASCs and dASCs both expressed SC markers p75 and GFAP but not S100. C, The amounts of NGF (above) and BDNF (below) in the conditioned medium from 48-h culture were measured by ELISA. The data are expressed as mean ± SEM, n = 3 experiments; ** P
    Figure Legend Snippet: A, Fibroblast-like ASCs (left) changed to bipolar, spindle-shaped dASCs (middle), which were similar to SCs (right). Scale bar: 50 µm. B, Western blotting analysis of cell lysates showed that ASCs and dASCs both expressed SC markers p75 and GFAP but not S100. C, The amounts of NGF (above) and BDNF (below) in the conditioned medium from 48-h culture were measured by ELISA. The data are expressed as mean ± SEM, n = 3 experiments; ** P

    Techniques Used: Western Blot, Enzyme-linked Immunosorbent Assay

    11) Product Images from "Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells"

    Article Title: Induction of Thermal Hyperalgesia and Synaptic Long-Term Potentiation in the Spinal Cord Lamina I by TNF-α and IL-1β is Mediated by Glial Cells

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5087-12.2013

    HFS activates microglia and astrocytes. Representative experiments show the changes in microglial ( A–C ) and astrocytic ( D–F ) specific markers after HFS of primary afferent fibers. A , D , Immunoreactivity for the microglia-specific marker Iba1 ( A , n = 15 slices) and the astrocyte-specific marker GFAP ( D , n = 16 slices) in untreated slices. B , 30 min after HFS, Iba1 immunofluorescence is increased on the ipsilateral site (right) but not contralaterally to HFS ( n = 14 slices). C , 120 min after HFS, Iba1 levels are back to control again ( n = 14 slices). E , Thirty minutes after HFS, GFAP immunoreactivity is unchanged compared with control ( n = 16 slices). F , Two hours after HFS of primary afferents, GFAP shows increased immunoreactivity on the ipsilateral (right) and contralateral (left) side ( n = 16 slices). G , Quantification of Iba1 and GFAP intensity 30 and 120 min after HFS. * p
    Figure Legend Snippet: HFS activates microglia and astrocytes. Representative experiments show the changes in microglial ( A–C ) and astrocytic ( D–F ) specific markers after HFS of primary afferent fibers. A , D , Immunoreactivity for the microglia-specific marker Iba1 ( A , n = 15 slices) and the astrocyte-specific marker GFAP ( D , n = 16 slices) in untreated slices. B , 30 min after HFS, Iba1 immunofluorescence is increased on the ipsilateral site (right) but not contralaterally to HFS ( n = 14 slices). C , 120 min after HFS, Iba1 levels are back to control again ( n = 14 slices). E , Thirty minutes after HFS, GFAP immunoreactivity is unchanged compared with control ( n = 16 slices). F , Two hours after HFS of primary afferents, GFAP shows increased immunoreactivity on the ipsilateral (right) and contralateral (left) side ( n = 16 slices). G , Quantification of Iba1 and GFAP intensity 30 and 120 min after HFS. * p

    Techniques Used: Marker, Immunofluorescence

    Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.
    Figure Legend Snippet: Receptors for the pro-inflammatory cytokines TNF-α and IL-1β are expressed by lamina I neurons and glial cells. Double immunostaining shows colocalization of TNFR1 ( Aa , Ba , green) and TNFR2 ( Ca , Da , Ea , green) and IL1R ( Fa , Ga , Ha , green) with the neuronal marker NeuN ( Ab , Cb , Fb , red), the astrocytic marker GFAP ( Bb , Db , Gb , red), and the microglial marker Iba1 ( Eb , Hb , red) in spinal cord lamina I. Overlays reveal that lamina I neurons express TNFR1, TNFR2, and IL-1R. TNFR1 and IL-1R are also expressed by GFAP-positive cells in the spinal dorsal horn. There is weak costaining for the individual receptors and the microglial marker Iba1.

    Techniques Used: Double Immunostaining, Marker

    12) Product Images from "Astrocytes regulate adult hippocampal neurogenesis through ephrin-B signaling"

    Article Title: Astrocytes regulate adult hippocampal neurogenesis through ephrin-B signaling

    Journal: Nature neuroscience

    doi: 10.1038/nn.3212

    Fc-ephrin-B2 promoted the neuronal differentiation of NSCs in vitro . ( a,b ) Stimulation by Fc-Ephrin-B2 induced NSCs to undergo neuronal differentiation (βIII-Tubulin + or Tubb3 ) in a dose-responsive fashion as measured by immunocytochemistry and QPCR ( n = 3, experimental replicates). Glial fibrilary acidic protein (GFAP) staining was slightly increased with ephrin-B2, but no increase in expression was observed by QPCR. ( c ) Blockage of ephrin-B2 receptors, EphB2 and EphB4, during Fc-ephrin-B2 (10 µg/mL) stimulation revealed that EphB4 mediates Fc-ephrin-B2’s proneuronal effect on NSCs ( n = 3, experimental replicates). ANOVA plus a multi-variable Tukey-Kramer analysis was conducted, with * indicating P
    Figure Legend Snippet: Fc-ephrin-B2 promoted the neuronal differentiation of NSCs in vitro . ( a,b ) Stimulation by Fc-Ephrin-B2 induced NSCs to undergo neuronal differentiation (βIII-Tubulin + or Tubb3 ) in a dose-responsive fashion as measured by immunocytochemistry and QPCR ( n = 3, experimental replicates). Glial fibrilary acidic protein (GFAP) staining was slightly increased with ephrin-B2, but no increase in expression was observed by QPCR. ( c ) Blockage of ephrin-B2 receptors, EphB2 and EphB4, during Fc-ephrin-B2 (10 µg/mL) stimulation revealed that EphB4 mediates Fc-ephrin-B2’s proneuronal effect on NSCs ( n = 3, experimental replicates). ANOVA plus a multi-variable Tukey-Kramer analysis was conducted, with * indicating P

    Techniques Used: In Vitro, Immunocytochemistry, Real-time Polymerase Chain Reaction, Staining, Expressing

    Lineage tracing of ephrin-B2-induced NSC differentiation. ( a ) Time course where ( b,c ) initially 90.7 ± 1.79% of β-Gal + cells were Nestin + /Sox2 + , and 53.8 ± 5.49% were GFAP + along radial process (Type 1 NSCs). ( d,e ) By day 5, 38.7 ± 2.73% and 37.3 ± 4.51% of β-Gal + cells were Sox2 + /DCX + and DCX + /NeuroD1 + , respectively, in Fc-ephrin-B2 injected mice vs. 27.7 ± 5.16% and 13.8 ± 4.38% in Anti-Fc controls. (f) At day 14, 41.1 ± 4.86% of β-Gal + cells were NeuN + for Fc-ephrin-B2 vs. 25.6 ± 4.24% for controls. ( g,h ) Representative sections where β-Gal + and BrdU + cells proliferated and differentiated over 14 days. (i) Compared to Anti-Fc, Fc-ephrin-B2 decreased the number of single Type 1 (“1”, 8.01 ± 1.91% vs. 17.8 ± 1.93%) and Type 2a NSCs (“2a”, 2.51 ± 2.34% vs. 10.8 ± 3.22%) and increased single neuroblast or neurons (“N”, 49.2 ± 4.57% vs. 30.9 ± 2.28%). Furthermore, ephrin-B2 decreased the number of doublets containing a Type 1 (“1+X”, 5.95 ± 1.47% vs. 11.8 ± 2.03%) or Type 2 (“2a+X”, 7.37 ± 3.00% vs. 16.5 ± 6.03%) cell and increased neuroblasts or neuron doublets (“N+N”,16.3 ± 1.82% vs. 3.87 ± 1.65%). Cluster size (1.55 ± 0.03 vs. 1.54 ± 0.04) and overall β-Gal + cell numbers ( Supplementary Fig. 5 ) were indistinguishable in Fc-ephrin-B2 vs. control mice. Thus, ephrin-B2 signaling increases neuronal differentiation without altering proliferation. ** P
    Figure Legend Snippet: Lineage tracing of ephrin-B2-induced NSC differentiation. ( a ) Time course where ( b,c ) initially 90.7 ± 1.79% of β-Gal + cells were Nestin + /Sox2 + , and 53.8 ± 5.49% were GFAP + along radial process (Type 1 NSCs). ( d,e ) By day 5, 38.7 ± 2.73% and 37.3 ± 4.51% of β-Gal + cells were Sox2 + /DCX + and DCX + /NeuroD1 + , respectively, in Fc-ephrin-B2 injected mice vs. 27.7 ± 5.16% and 13.8 ± 4.38% in Anti-Fc controls. (f) At day 14, 41.1 ± 4.86% of β-Gal + cells were NeuN + for Fc-ephrin-B2 vs. 25.6 ± 4.24% for controls. ( g,h ) Representative sections where β-Gal + and BrdU + cells proliferated and differentiated over 14 days. (i) Compared to Anti-Fc, Fc-ephrin-B2 decreased the number of single Type 1 (“1”, 8.01 ± 1.91% vs. 17.8 ± 1.93%) and Type 2a NSCs (“2a”, 2.51 ± 2.34% vs. 10.8 ± 3.22%) and increased single neuroblast or neurons (“N”, 49.2 ± 4.57% vs. 30.9 ± 2.28%). Furthermore, ephrin-B2 decreased the number of doublets containing a Type 1 (“1+X”, 5.95 ± 1.47% vs. 11.8 ± 2.03%) or Type 2 (“2a+X”, 7.37 ± 3.00% vs. 16.5 ± 6.03%) cell and increased neuroblasts or neuron doublets (“N+N”,16.3 ± 1.82% vs. 3.87 ± 1.65%). Cluster size (1.55 ± 0.03 vs. 1.54 ± 0.04) and overall β-Gal + cell numbers ( Supplementary Fig. 5 ) were indistinguishable in Fc-ephrin-B2 vs. control mice. Thus, ephrin-B2 signaling increases neuronal differentiation without altering proliferation. ** P

    Techniques Used: Injection, Mouse Assay

    In vivo , SGZ Type 2a NSCs, Type 2b neuronal precursors, and Type 3 neuroblasts express EphB4, and hippocampal astrocytes express ephrin-B2. ( a ) Staining of the hippocampal dentate gyrus showed that GFAP + hilar (H) astrocytes express ephrin-B2. In addition, cells in the SGZ and neurons in the granule cell layer (GCL) express EphB4 + on the cell soma. Scale bar represents 100 µm. ( b ) GFAP + astrocytes adjacent to the SGZ co-express ephrin-B2. ( c,d ) EphB4 expression persists throughout NSC neuronal differentiation, including Type 2a NSCs (Sox2 + /DCX − /GFAP − ), Type 2b neuronal precursors (Sox2 + /DCX + ), and Type 3 neuroblasts (DCX + ). ( c ) Confocal images show EphB4 expression in Sox2 + /DCX − cells (large arrowheads) in the SGZ. Magnified region depicts the presence of EphB4 expression on a Sox2 + /DCX + Type 2b neuronal precursor (small arrowhead) and a DCX + Type 3 neuroblast (arrow). ( d ) Confocal images also show EphB4 expression by Sox2 + /GFAP − cells (small arrowheads); therefore, Sox2 + /DCX − /GFAP − Type 2a NSCs also express EphB4. ( e ) Given the close proximity of ephrin-B2 + astrocytes to EphB4 + cells in the SGZ, ephrin-B2/EphB4 juxtacrine signaling is in a position to induce NSC differentiation into ( f ) Sox2 + /DCX + Type 2b neuronal precursors (small arrowhead) and subsequently Sox2 − /DCX + Type 3 neuroblasts (arrow). The scale bars represent 10 µm.
    Figure Legend Snippet: In vivo , SGZ Type 2a NSCs, Type 2b neuronal precursors, and Type 3 neuroblasts express EphB4, and hippocampal astrocytes express ephrin-B2. ( a ) Staining of the hippocampal dentate gyrus showed that GFAP + hilar (H) astrocytes express ephrin-B2. In addition, cells in the SGZ and neurons in the granule cell layer (GCL) express EphB4 + on the cell soma. Scale bar represents 100 µm. ( b ) GFAP + astrocytes adjacent to the SGZ co-express ephrin-B2. ( c,d ) EphB4 expression persists throughout NSC neuronal differentiation, including Type 2a NSCs (Sox2 + /DCX − /GFAP − ), Type 2b neuronal precursors (Sox2 + /DCX + ), and Type 3 neuroblasts (DCX + ). ( c ) Confocal images show EphB4 expression in Sox2 + /DCX − cells (large arrowheads) in the SGZ. Magnified region depicts the presence of EphB4 expression on a Sox2 + /DCX + Type 2b neuronal precursor (small arrowhead) and a DCX + Type 3 neuroblast (arrow). ( d ) Confocal images also show EphB4 expression by Sox2 + /GFAP − cells (small arrowheads); therefore, Sox2 + /DCX − /GFAP − Type 2a NSCs also express EphB4. ( e ) Given the close proximity of ephrin-B2 + astrocytes to EphB4 + cells in the SGZ, ephrin-B2/EphB4 juxtacrine signaling is in a position to induce NSC differentiation into ( f ) Sox2 + /DCX + Type 2b neuronal precursors (small arrowhead) and subsequently Sox2 − /DCX + Type 3 neuroblasts (arrow). The scale bars represent 10 µm.

    Techniques Used: In Vivo, Staining, Expressing

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    Article Snippet: .. After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer. .. Coverslips were mounted onto slides with ProLong Gold Media (Invitrogen) and visualized with Zeiss AxioImage Olympus FV-1000 confocal microscope (Olympus America Inc.) and images captured with FluoView v. 5.0 software (Olympus America Inc.).

    Article Title: Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-?12,14-prostaglandin J2
    Article Snippet: To show the specificity of the PGDS staining, the antibody was first pre-incubated with the PGDS blocking peptide (w/w, Cayman) for 1 h at room temperature. .. After PGDS staining, primary culture was incubated with mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (1:100) overnight at 4°C followed by donkey anti-mouse IgG conjugated with Alexa Fluor 488 fluorescent dye (1:500; Molecular Probes).

    Plasmid Purification:

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: For immunohistochemical studies, the antibodies used were anti-CD106 (VCAM1, mouse-IgG1, 1.4C3; Acris), CD62P (P-selectin, mouse IgG2a, 1E3; Santa Cruz Biotechnology, Inc.), and PLP (Proteo lipid protein, mouse-IgG2a, plpc1; AbD Serotec). .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Incubation:

    Article Title: Free-Radical Scavenger Edaravone Treatment Confers Neuroprotection Against Traumatic Brain Injury in Rats
    Article Snippet: Immunohistochemical stains for light microscopy were performed using antibodies for the following proteins: glial fibrillary acidic protein (GFAP, 1:200 monoclonal mouse-anti-GFAP, ThermoFisher Scientific, Waltham, MA), vimentin (1:100 monoclonal mouse-anti-vimentin, ThermoFisher Scientific), S-100 (1:200 monoclonal mouse-anti-S-100, Chemicon, Billerica, MA), OX42 (1:200 monoclonal mouse-anti-S-100, Chemicon) and activated caspase-3 (1:100 polyclonal rabbit anti-cleaved caspase-3 [Asp175], Bioworld Technology Inc., Louis Park, MN). .. Endogenous peroxidase was blocked with 0.3% H2 O2 in methanol and was followed by incubation in blocking buffer (10% normal goat serum and 0.3% Triton-X in TBS) for 30 min.

    Article Title: Stathmin-Deficient Mice Develop an Age-Dependent Axonopathy of the Central and Peripheral Nervous Systems
    Article Snippet: Either 10-μm frozen or 1-μm epoxy sections were incubated with primary antibody and developed using the Vectastain ABC kit (Vector Laboratories, Burlingame, CA). .. The following primary antibodies were used: rabbit anti-stathmin C-terminal peptide, 1:1000; mouse anti-glial fibrillary acidic protein (GFAP) (Zymed, San Francisco, CA), 1:200; mouse anti-NFH-(strongly phosphorylated) SMI 35 (Sternberger Monoclonals, Baltimore, MD), 1:5000; mouse anti-NFH-(nonphosphorylated) SMI 32 (Sternberger Monoclonals), 1:2000; mouse anti-NFH-(intermediately phosphorylated) SMI 31 (Sternberger Monoclonals), 1:5000; mouse anti-NFL (Sigma Chemical Co., St. Louis, MO), 1:100.

    Article Title: The Impact of Alpha-Syntrophin Deletion on the Changes in Tissue Structure and Extracellular Diffusion Associated with Cell Swelling under Physiological and Pathological Conditions
    Article Snippet: .. The slices were then incubated with the primary mouse antibody against glial fibrillary acidic protein (anti-GFAP coupled to Alexa 488;1∶800) at 4°C overnight (eBioscience, San Diego, CA, USA) and mounted using Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA). .. The tissue slices were then examined using an LSM 5 DUO spectral confocal microscope equipped with Arg/HeNe lasers and 40× or 63× oil objectives (Zeiss, Germany).

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: .. Primary antibodies: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶500, polyclonal antibody; Santa-cruz), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), and anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam) diluted in antibody diluents solution (Invitrogen, Carlsbd, CA) were added to the tissue section samples and incubated at 4°C overnight. ..

    Article Title: Segregation of expression of mPeriod gene homologs in neurons and glia: possible divergent roles of mPeriod1 and mPeriod2 in the brain
    Article Snippet: For immunofluorescent labeling, thin (40 µm), free-floating brain sections were washed five times in PBST (PBS with 0.1% Triton X-100), blocked for 1 h in 5% horse serum/PBST, and incubated (overnight, 4°C) in primary antibodies. .. The following primary antibodies were used: rabbit polyclonal anti-GFP (1:2000; Eusera, Edmonton, Alberta, Canada), goat polyclonal anti-GFP (1:2000; Eusera), rabbit polyclonal anti-DsRED (1:200; Clontech), mouse monoclonal anti-NeuN (1:1000; Chemicon, Temecula, CA, USA), mouse monoclonal anti-GFAP (1:500; Molecular Probes, Eugene, OR, USA), goat polyclonal anti-SOX-2 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-doublecortin (1:1000; Santa Cruz Biotechnology), guinea pig polyclonal anti-AVP (1:4000; Peninsula Laboratories, San Carlos, CA, USA) and rabbit polyclonal anti-calbindin (1:400; Millipore, Billerica, MA, USA).

    Article Title: Novel botanical drug DA-9803 prevents deficits in Alzheimer’s mouse models
    Article Snippet: .. The coronal sections were then permeabilized with Triton X-100, blocked with normal goat serum (NGS), and incubated with the various primary antibodies: 6E10 (monoclonal 6E10, 1:500; Covance), IBA1 (rabbit anti-Iba1, 1:2000; Wako), glial fibrillary acidic protein (GFAP) (mouse anti-GFAP, 1:200; Thermo Scientific), and MAP2 (mouse anti-MAP2, 1:100; Abcam) for 2 hours at room temperature. .. The coronal sections were then incubated with the respective secondary antibodies for 1 hour and mounted with ProLong Antifade reagent (Invitrogen).

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore). .. Then the slides were washed in PBS three times and were incubated with appropriate secondary antibodies conjugated either with fluorescein isothiocyanate ( FITC) (1∶500, Chemicon) or rhodamine (1∶500, Chemicon) at 37°C for 2 hours.

    Article Title: Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury
    Article Snippet: .. After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer. .. Coverslips were mounted onto slides with ProLong Gold Media (Invitrogen) and visualized with Zeiss AxioImage Olympus FV-1000 confocal microscope (Olympus America Inc.) and images captured with FluoView v. 5.0 software (Olympus America Inc.).

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: These cultures were infected with either AcVSVG-CAGFP or AcRVG-CAGFP at an MOI of 100 at 8 days after incubation. .. Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C.

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: Tissue was incubated in primary antibody, 5% NGS, and 0.5% TritonX100 in 0.1M PB overnight at 4°C, washed three times for 10 minutes in 0.1M PB, and incubated in secondary antibody, 5% NGS, and 0.5% TritonX100 in 0.1M PB for 60–120 minutes at room temperature. .. The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461).

    Article Title: Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-?12,14-prostaglandin J2
    Article Snippet: .. After PGDS staining, primary culture was incubated with mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (1:100) overnight at 4°C followed by donkey anti-mouse IgG conjugated with Alexa Fluor 488 fluorescent dye (1:500; Molecular Probes). .. Following washes, stained samples were observed and acquired with a microscope IX 50 (Olympus) or with the Axiovert 200M microscope coupled to Apotome (Zeiss, Göttingen, Germany).

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: .. Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377). .. The next day, tissue was washed three times for 10 min in 0.1 M PB (pH = 7.4) and then incubated at room temperature in 5% NGS, 0.5% Triton-X100, and secondary antibody for 2 h. Secondary antibodies used for visualization were conjugated with Alexa-Fluor dyes (Alexa-488, 1:1000, Invitrogen, ThermoFisher Scientific, Cat#A-l 1001).

    Immunocytochemistry:

    Article Title: Stathmin-Deficient Mice Develop an Age-Dependent Axonopathy of the Central and Peripheral Nervous Systems
    Article Snippet: Paragraph title: Immunocytochemistry ... The following primary antibodies were used: rabbit anti-stathmin C-terminal peptide, 1:1000; mouse anti-glial fibrillary acidic protein (GFAP) (Zymed, San Francisco, CA), 1:200; mouse anti-NFH-(strongly phosphorylated) SMI 35 (Sternberger Monoclonals, Baltimore, MD), 1:5000; mouse anti-NFH-(nonphosphorylated) SMI 32 (Sternberger Monoclonals), 1:2000; mouse anti-NFH-(intermediately phosphorylated) SMI 31 (Sternberger Monoclonals), 1:5000; mouse anti-NFL (Sigma Chemical Co., St. Louis, MO), 1:100.

    Article Title: Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury
    Article Snippet: Paragraph title: Oxygen–glucose deprivation and immunocytochemistry ... After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer.

    Activity Assay:

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: Antigen retrieval was done using EnVision FLEX Target Retrieval Solution, Low pH (DAKO), and endogenous peroxidase activity was blocked with 3% hydrogen peroxide in methanol. .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Mass Spectrometry:

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: .. The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461). .. To visualize IL-1β and GFAP, Alexa Fluor conjugated secondary antibodies, goat anti-rabbit 488 (1:1000, ThermoFisher Scientific, Waltham, MA, Cat # ) and goat anti-GFAP Dylight 405 (1:1000, ThermoFisher Scientific, Waltham, MA, Cat # 35501BID) were used.

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: For histochemical studies, 4-µm-thick formalin fixed paraffin embedded (FFPE) tissues sections from three MS patients and three patients without inflammatory demyelinating CNS disease were stained with luxol fast blue for myelin integrity and H & E for inflammatory infiltrates. .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: .. Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377). .. The next day, tissue was washed three times for 10 min in 0.1 M PB (pH = 7.4) and then incubated at room temperature in 5% NGS, 0.5% Triton-X100, and secondary antibody for 2 h. Secondary antibodies used for visualization were conjugated with Alexa-Fluor dyes (Alexa-488, 1:1000, Invitrogen, ThermoFisher Scientific, Cat#A-l 1001).

    Modification:

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: Cerebellar cultures were maintained in Dulbecco's modified Eagle's medium-F12 medium (Gibco Laboratories) containing 0.5% FCS, putrescine (100 μM), sodium selenite (30 nM), l -glutamine (4 mM), triiodothyronine (0.5 μg/ml), progesterone (5 nM), bovine insulin (10 μg/ml), transferrin (100 μg/ml), and gentamicin (10 μg/ml). .. Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C.

    Derivative Assay:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F. .. For the quantification of hNSC differentiation, representative hippocampal sections from 4 to 6 distinct transplant sites derived from 4 engrafted animals were subjected to dual immunofluorescence staining.

    Immunohistochemistry:

    Article Title: Free-Radical Scavenger Edaravone Treatment Confers Neuroprotection Against Traumatic Brain Injury in Rats
    Article Snippet: .. Immunohistochemical stains for light microscopy were performed using antibodies for the following proteins: glial fibrillary acidic protein (GFAP, 1:200 monoclonal mouse-anti-GFAP, ThermoFisher Scientific, Waltham, MA), vimentin (1:100 monoclonal mouse-anti-vimentin, ThermoFisher Scientific), S-100 (1:200 monoclonal mouse-anti-S-100, Chemicon, Billerica, MA), OX42 (1:200 monoclonal mouse-anti-S-100, Chemicon) and activated caspase-3 (1:100 polyclonal rabbit anti-cleaved caspase-3 [Asp175], Bioworld Technology Inc., Louis Park, MN). .. Endogenous peroxidase was blocked with 0.3% H2 O2 in methanol and was followed by incubation in blocking buffer (10% normal goat serum and 0.3% Triton-X in TBS) for 30 min.

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Paragraph title: Immunohistochemistry, stereology, and confocal microscopy ... For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: The Impact of Alpha-Syntrophin Deletion on the Changes in Tissue Structure and Extracellular Diffusion Associated with Cell Swelling under Physiological and Pathological Conditions
    Article Snippet: Paragraph title: Immunohistochemistry ... The slices were then incubated with the primary mouse antibody against glial fibrillary acidic protein (anti-GFAP coupled to Alexa 488;1∶800) at 4°C overnight (eBioscience, San Diego, CA, USA) and mounted using Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA).

    Article Title: Segregation of expression of mPeriod gene homologs in neurons and glia: possible divergent roles of mPeriod1 and mPeriod2 in the brain
    Article Snippet: Paragraph title: Immunofluorescent and immunohistochemical detection ... The following primary antibodies were used: rabbit polyclonal anti-GFP (1:2000; Eusera, Edmonton, Alberta, Canada), goat polyclonal anti-GFP (1:2000; Eusera), rabbit polyclonal anti-DsRED (1:200; Clontech), mouse monoclonal anti-NeuN (1:1000; Chemicon, Temecula, CA, USA), mouse monoclonal anti-GFAP (1:500; Molecular Probes, Eugene, OR, USA), goat polyclonal anti-SOX-2 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-doublecortin (1:1000; Santa Cruz Biotechnology), guinea pig polyclonal anti-AVP (1:4000; Peninsula Laboratories, San Carlos, CA, USA) and rabbit polyclonal anti-calbindin (1:400; Millipore, Billerica, MA, USA).

    Article Title: Novel botanical drug DA-9803 prevents deficits in Alzheimer’s mouse models
    Article Snippet: Paragraph title: Immunohistochemistry ... The coronal sections were then permeabilized with Triton X-100, blocked with normal goat serum (NGS), and incubated with the various primary antibodies: 6E10 (monoclonal 6E10, 1:500; Covance), IBA1 (rabbit anti-Iba1, 1:2000; Wako), glial fibrillary acidic protein (GFAP) (mouse anti-GFAP, 1:200; Thermo Scientific), and MAP2 (mouse anti-MAP2, 1:100; Abcam) for 2 hours at room temperature.

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: Paragraph title: 2.3.2. Immunohistochemistry ... The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461).

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: Paragraph title: Immunohistochemistry. ... For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: Paragraph title: 2.6. Immunohistochemistry ... Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377).

    Infection:

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: Paragraph title: Primary cell cultures and infection with recombinant baculoviruses. ... Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C.

    Light Microscopy:

    Article Title: Free-Radical Scavenger Edaravone Treatment Confers Neuroprotection Against Traumatic Brain Injury in Rats
    Article Snippet: .. Immunohistochemical stains for light microscopy were performed using antibodies for the following proteins: glial fibrillary acidic protein (GFAP, 1:200 monoclonal mouse-anti-GFAP, ThermoFisher Scientific, Waltham, MA), vimentin (1:100 monoclonal mouse-anti-vimentin, ThermoFisher Scientific), S-100 (1:200 monoclonal mouse-anti-S-100, Chemicon, Billerica, MA), OX42 (1:200 monoclonal mouse-anti-S-100, Chemicon) and activated caspase-3 (1:100 polyclonal rabbit anti-cleaved caspase-3 [Asp175], Bioworld Technology Inc., Louis Park, MN). .. Endogenous peroxidase was blocked with 0.3% H2 O2 in methanol and was followed by incubation in blocking buffer (10% normal goat serum and 0.3% Triton-X in TBS) for 30 min.

    Immunostaining:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: For stereologic quantification, every 10th section through the entire hippocampus was processed for BrdUrd immunostaining (mouse anti-BrdUrd, clone AH4H7-1/131-14871, 1:200; Millipore). .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: Paragraph title: 3, 3′- diaminobenzidine Tetra Hydrochloride (DAB) Immunostaining for CAST Analysis ... Primary antibodies: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶500, polyclonal antibody; Santa-cruz), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), and anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam) diluted in antibody diluents solution (Invitrogen, Carlsbd, CA) were added to the tissue section samples and incubated at 4°C overnight.

    Recombinant:

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: Paragraph title: Primary cell cultures and infection with recombinant baculoviruses. ... Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C.

    Immunofluorescence:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F. .. Worley, Johns Hopkins University, Baltimore, MD).

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: Paragraph title: Immunofluorescence Studies Using Confocal Microscopy ... The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore).

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako). .. The secondary antibodies used were goat anti–rabbit-HRP (IgG; Life Technologies), donkey anti–mouse-568 (IgG; Invitrogen), and streptavidin-488 (Invitrogen).

    Article Title: Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-?12,14-prostaglandin J2
    Article Snippet: Paragraph title: Immunofluorescence staining ... After PGDS staining, primary culture was incubated with mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (1:100) overnight at 4°C followed by donkey anti-mouse IgG conjugated with Alexa Fluor 488 fluorescent dye (1:500; Molecular Probes).

    Flow Cytometry:

    Article Title: Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury
    Article Snippet: The cultures were then placed in an airtight incubation chamber (CBS Scientific) and flushed with a continuous influx of 1% O2 at a flow rate of 20 L/minute. .. After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer.

    Labeling:

    Article Title: Segregation of expression of mPeriod gene homologs in neurons and glia: possible divergent roles of mPeriod1 and mPeriod2 in the brain
    Article Snippet: For immunofluorescent labeling, thin (40 µm), free-floating brain sections were washed five times in PBST (PBS with 0.1% Triton X-100), blocked for 1 h in 5% horse serum/PBST, and incubated (overnight, 4°C) in primary antibodies. .. The following primary antibodies were used: rabbit polyclonal anti-GFP (1:2000; Eusera, Edmonton, Alberta, Canada), goat polyclonal anti-GFP (1:2000; Eusera), rabbit polyclonal anti-DsRED (1:200; Clontech), mouse monoclonal anti-NeuN (1:1000; Chemicon, Temecula, CA, USA), mouse monoclonal anti-GFAP (1:500; Molecular Probes, Eugene, OR, USA), goat polyclonal anti-SOX-2 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-doublecortin (1:1000; Santa Cruz Biotechnology), guinea pig polyclonal anti-AVP (1:4000; Peninsula Laboratories, San Carlos, CA, USA) and rabbit polyclonal anti-calbindin (1:400; Millipore, Billerica, MA, USA).

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: .. Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C. .. After incubation, cells were washed with PBS and incubated with Alexa 568-conjugated anti-mouse or Alexa 488-conjugated anti-rabbit immunoglobulin antibody (Molecular Probes) for 30 min at room temperature.

    Avidin-Biotin Assay:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Color was developed using the avidin-biotin-complex method and enhanced diaminobenzidine substrate (Vector Labs), and all sections were counterstained with hematoxylin. .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Microscopy:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Stereologic enumeration was conducted using an Olympus BX60 microscope equipped with an MBF CX9000 color digital camera, 100 × (oil-immersion, 1.30NA) objective lens, 3-axis motorized stage and StereoInvestigator software (MBF Biosciences, v9). .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: The Impact of Alpha-Syntrophin Deletion on the Changes in Tissue Structure and Extracellular Diffusion Associated with Cell Swelling under Physiological and Pathological Conditions
    Article Snippet: The slices were then incubated with the primary mouse antibody against glial fibrillary acidic protein (anti-GFAP coupled to Alexa 488;1∶800) at 4°C overnight (eBioscience, San Diego, CA, USA) and mounted using Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA). .. The tissue slices were then examined using an LSM 5 DUO spectral confocal microscope equipped with Arg/HeNe lasers and 40× or 63× oil objectives (Zeiss, Germany).

    Article Title: Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury
    Article Snippet: After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer. .. Coverslips were mounted onto slides with ProLong Gold Media (Invitrogen) and visualized with Zeiss AxioImage Olympus FV-1000 confocal microscope (Olympus America Inc.) and images captured with FluoView v. 5.0 software (Olympus America Inc.).

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C. .. Images were acquired on a Zeiss LSM510 confocal microscope (Carl Zeiss Inc., Thornwood, N.Y.).

    Article Title: Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-?12,14-prostaglandin J2
    Article Snippet: After PGDS staining, primary culture was incubated with mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (1:100) overnight at 4°C followed by donkey anti-mouse IgG conjugated with Alexa Fluor 488 fluorescent dye (1:500; Molecular Probes). .. Following washes, stained samples were observed and acquired with a microscope IX 50 (Olympus) or with the Axiovert 200M microscope coupled to Apotome (Zeiss, Göttingen, Germany).

    Confocal Microscopy:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Paragraph title: Immunohistochemistry, stereology, and confocal microscopy ... For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: Paragraph title: Immunofluorescence Studies Using Confocal Microscopy ... The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: .. The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat016-26461). .. To visualize IL-1β and GFAP, Alexa Fluor conjugated secondary antibodies, goat anti-rabbit 488 (1:1000, ThermoFisher Scientific, Waltham, MA, Cat # ) and goat anti-GFAP Dylight 405 (1:1000, ThermoFisher Scientific, Waltham, MA, Cat # 35501BID) were used.

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377). .. The next day, tissue was washed three times for 10 min in 0.1 M PB (pH = 7.4) and then incubated at room temperature in 5% NGS, 0.5% Triton-X100, and secondary antibody for 2 h. Secondary antibodies used for visualization were conjugated with Alexa-Fluor dyes (Alexa-488, 1:1000, Invitrogen, ThermoFisher Scientific, CatA-l 1001).

    Purification:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F. .. Worley, Johns Hopkins University, Baltimore, MD).

    Plasmid Preparation:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F. .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: Primary antibodies: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶500, polyclonal antibody; Santa-cruz), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), and anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam) diluted in antibody diluents solution (Invitrogen, Carlsbd, CA) were added to the tissue section samples and incubated at 4°C overnight. .. Tissue sections were washed again 3 times for 3 min each with PBS and stained with 3, 3′ - diamino-benzidine tetra hydrochloride (DAB, sigma, St. Louis, MO, USA) and SG substrate kit (Vector, Burlingame, CA, USA) for 5 min at room temperature.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore). .. The slides were stained with 4′-6-diamidino-2-phenylindole (DAPI) (Vector, Burlingame, USA) to counterstain the nuclei and were coverslipped with mounting medium.

    Article Title: In Vitro and In Vivo Gene Delivery by Recombinant Baculoviruses
    Article Snippet: Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C. .. Cells were then double labeled with either rabbit anti-GFP (Molecular Probes, Eugene, Oreg.), mouse anti-Calbindin (Swant, Bellinzona, Switzerland), mouse anti-MAP2 (ICN Biomedicals, Costa Mesa, Calif.), or mouse anti- glial fibrillary acidic protein (GFAP) (Zymed Laboratories, Inc., South San Francisco, Calif.) for 30 min to 1 h at 37°C.

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377). .. Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377).

    Software:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Stereologic enumeration was conducted using an Olympus BX60 microscope equipped with an MBF CX9000 color digital camera, 100 × (oil-immersion, 1.30NA) objective lens, 3-axis motorized stage and StereoInvestigator software (MBF Biosciences, v9). .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Article Title: Spatial distribution of insulin-like growth factor binding protein-2 following hypoxic-ischemic injury
    Article Snippet: After blocking for 1 h at room temperature, the cells were incubated in primary antibodies (IGFBP-2, R & D Systems, AF797; MAP2, Sigma, M9942; GFAP, Sigma, G6171) and secondary antibodies (Alexa Fluors, 1:1000, Invitrogen) in blocking buffer. .. Coverslips were mounted onto slides with ProLong Gold Media (Invitrogen) and visualized with Zeiss AxioImage Olympus FV-1000 confocal microscope (Olympus America Inc.) and images captured with FluoView v. 5.0 software (Olympus America Inc.).

    Next-Generation Sequencing:

    Article Title: Novel botanical drug DA-9803 prevents deficits in Alzheimer’s mouse models
    Article Snippet: .. The coronal sections were then permeabilized with Triton X-100, blocked with normal goat serum (NGS), and incubated with the various primary antibodies: 6E10 (monoclonal 6E10, 1:500; Covance), IBA1 (rabbit anti-Iba1, 1:2000; Wako), glial fibrillary acidic protein (GFAP) (mouse anti-GFAP, 1:200; Thermo Scientific), and MAP2 (mouse anti-MAP2, 1:100; Abcam) for 2 hours at room temperature. .. The coronal sections were then incubated with the respective secondary antibodies for 1 hour and mounted with ProLong Antifade reagent (Invitrogen).

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: Tissue was incubated in primary antibody, 5% NGS, and 0.5% TritonX100 in 0.1M PB overnight at 4°C, washed three times for 10 minutes in 0.1M PB, and incubated in secondary antibody, 5% NGS, and 0.5% TritonX100 in 0.1M PB for 60–120 minutes at room temperature. .. The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461).

    Article Title: Dorsal hippocampal neural immune signaling regulates heroin-conditioned immunomodulation but not heroin-conditioned place preference
    Article Snippet: .. Tissue was then incubated overnight at 4°C in 5% NGS, 0.5% Triton-X100, and primary antibody, mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat# MS-1376P) or mouse anti-NeuN (1:1000, Millipore, Burlington, MA, Cat# MAB377). .. The next day, tissue was washed three times for 10 min in 0.1 M PB (pH = 7.4) and then incubated at room temperature in 5% NGS, 0.5% Triton-X100, and secondary antibody for 2 h. Secondary antibodies used for visualization were conjugated with Alexa-Fluor dyes (Alexa-488, 1:1000, Invitrogen, ThermoFisher Scientific, Cat#A-l 1001).

    Sampling:

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: Sampling parameters (grid and counting frame size) were empirically determined to achieve low coefficients of error ( < 0.031 ± 0.002; n = 8) for hippocampus. .. For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F.

    Formalin-fixed Paraffin-Embedded:

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: For histochemical studies, 4-µm-thick formalin fixed paraffin embedded (FFPE) tissues sections from three MS patients and three patients without inflammatory demyelinating CNS disease were stained with luxol fast blue for myelin integrity and H & E for inflammatory infiltrates. .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Marker:

    Article Title: Segregation of expression of mPeriod gene homologs in neurons and glia: possible divergent roles of mPeriod1 and mPeriod2 in the brain
    Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-GFP (1:2000; Eusera, Edmonton, Alberta, Canada), goat polyclonal anti-GFP (1:2000; Eusera), rabbit polyclonal anti-DsRED (1:200; Clontech), mouse monoclonal anti-NeuN (1:1000; Chemicon, Temecula, CA, USA), mouse monoclonal anti-GFAP (1:500; Molecular Probes, Eugene, OR, USA), goat polyclonal anti-SOX-2 (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), goat polyclonal anti-doublecortin (1:1000; Santa Cruz Biotechnology), guinea pig polyclonal anti-AVP (1:4000; Peninsula Laboratories, San Carlos, CA, USA) and rabbit polyclonal anti-calbindin (1:400; Millipore, Billerica, MA, USA). .. In some experiments, sections were counterstained with the nuclear marker TO-PRO® -3 (1:1000; Invitrogen).

    Staining:

    Article Title: Free-Radical Scavenger Edaravone Treatment Confers Neuroprotection Against Traumatic Brain Injury in Rats
    Article Snippet: Paragraph title: Nissl staining and immunohistochemistry ... Immunohistochemical stains for light microscopy were performed using antibodies for the following proteins: glial fibrillary acidic protein (GFAP, 1:200 monoclonal mouse-anti-GFAP, ThermoFisher Scientific, Waltham, MA), vimentin (1:100 monoclonal mouse-anti-vimentin, ThermoFisher Scientific), S-100 (1:200 monoclonal mouse-anti-S-100, Chemicon, Billerica, MA), OX42 (1:200 monoclonal mouse-anti-S-100, Chemicon) and activated caspase-3 (1:100 polyclonal rabbit anti-cleaved caspase-3 [Asp175], Bioworld Technology Inc., Louis Park, MN).

    Article Title: Human Neural Stem Cell Transplantation Ameliorates Radiation-Induced Cognitive Dysfunction
    Article Snippet: For dual immunofluorescence analyses, we utilized anti-BrdUrd (purified rat, 1:200; Millipore), anti-neuron–specific nuclear antigen (anti-NeuN; mouse, 1:250; Millipore), anti-glial fibrillary acidic protein (anti-GFAP; mouse, 1:500; Invitrogen), antinuclei (human specific antinuclei, HuNu, mouse clone 235-1, 1:150; Millipore), nestin (mouse, 1:200; Millipore), Ki67 (rabbit, 1:150; Millipore), S100ß protein (S100, 1:200, rabbit; Millipore), and ß-III-tubulin (Tuj1, rabbit, 1:200; Covance) antibodies and polyclonal rabbit anti-Arc antibody (1:500; a kind gift from Dr. P.F. .. For the quantification of hNSC differentiation, representative hippocampal sections from 4 to 6 distinct transplant sites derived from 4 engrafted animals were subjected to dual immunofluorescence staining.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: Primary antibodies: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶500, polyclonal antibody; Santa-cruz), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), and anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam) diluted in antibody diluents solution (Invitrogen, Carlsbd, CA) were added to the tissue section samples and incubated at 4°C overnight. .. Tissue sections were washed again 3 times for 3 min each with PBS and stained with 3, 3′ - diamino-benzidine tetra hydrochloride (DAB, sigma, St. Louis, MO, USA) and SG substrate kit (Vector, Burlingame, CA, USA) for 5 min at room temperature.

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells
    Article Snippet: The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore). .. The slides were stained with 4′-6-diamidino-2-phenylindole (DAPI) (Vector, Burlingame, USA) to counterstain the nuclei and were coverslipped with mounting medium.

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β
    Article Snippet: For tissue stained with anti-Iba-1 antibody, tissue was incubated in endogenous biotin and streptavidin blocks (Vector Laboratories, Burlingame, CA, USA) for 30 minutes each at room temperature, according to the manufacturer’s instructions. .. The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461).

    Article Title: VLA-4 blockade promotes differential routes into human CNS involving PSGL-1 rolling of T cells and MCAM-adhesion of TH17 cells
    Article Snippet: For histochemical studies, 4-µm-thick formalin fixed paraffin embedded (FFPE) tissues sections from three MS patients and three patients without inflammatory demyelinating CNS disease were stained with luxol fast blue for myelin integrity and H & E for inflammatory infiltrates. .. For immunofluorescence studies, the primary antibodies used were CD146 (MCAM, rabbit monoclonal IgG, EPR3208; Millipore), CD8 (mouse monoclonal, IgG1, 144B; Abcam), rabbit anti-laminin α4 , GFAP (mouse monoclonal IgG1, 131-17719; Invitrogen) and CD68 (mouse monoclonal IgG3k, PG-M1; Dako).

    Article Title: Enteric glia modulate epithelial cell proliferation and differentiation through 15-deoxy-?12,14-prostaglandin J2
    Article Snippet: .. After PGDS staining, primary culture was incubated with mouse monoclonal anti-glial fibrillary acidic protein (GFAP) (1:100) overnight at 4°C followed by donkey anti-mouse IgG conjugated with Alexa Fluor 488 fluorescent dye (1:500; Molecular Probes). .. Following washes, stained samples were observed and acquired with a microscope IX 50 (Olympus) or with the Axiovert 200M microscope coupled to Apotome (Zeiss, Göttingen, Germany).

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  • 99
    Thermo Fisher mouse anti gfap
    <t>IL-1β</t> signal is colocalized with <t>GFAP,</t> and not with Iba-1 or NeuN, in the dorsal hippocampus in stressed and non-stressed animals A. Representative images of IL-1β, NeuN, Iba-1, and GFAP immunoreactivity in the dentate gyrus of the DH (AP −3.36 mm from bregma) acquired at 20X are shown. Because we did not detect any differences in colocalization between stressed and non-stressed rats, all images here are taken from animals that received stress exposure. Bitplane Imaris was used for background subtraction to better visualize individual cells presented. B. Bitplane Imaris software was used to calculate the colocalization of the IL-1β signal with GFAP, Iba-1, and NeuN. Colocalization analyses revealed that the percent of the IL-1β signal colocalized with GFAP was significantly greater than the percent of the IL-1β signal colocalized with either Iba-1 or NeuN. * p
    Mouse Anti Gfap, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti gfap/product/Thermo Fisher
    Average 99 stars, based on 31 article reviews
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    mouse anti gfap - by Bioz Stars, 2020-04
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    94
    Thermo Fisher rat monoclonal anti gfap
    Neonatal intravenous injection of AAV8.EFS.GFP enables neuronal transduction. a Brain imaged with fluorescence microscope in CD-1 pups injected intravenously with AAV8.EFS. GFP (GFP) and uninjected controls. b , c Representative images of GFP immunostaining in brain at b low and c at higher magnifications in mice injected with the GFP vector and uninjected controls show a decreasing rostro-caudal gradient with preferential transduction of forebrain and midbrain. d Computational quantification of GFP immunostaining showed a significant increase in AAV8.EFS. GFP -injected versus uninjected littermates ( n = 4). e Immunofluorescence of cortical staining for DAPI (blue), GFP (green), and NeuN, <t>GFAP,</t> Olig-2, CD68 (red) identifying neurons, astrocytes, oligodendrocytes and microglial cells, respectively. f Colocalisation measured by Pearson’s coefficient showed a restricted neuronal transduction. Horizontal lines display the mean ± SEM. d Unpaired two-tailed Student’s t test * p
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    Image Search Results


    IL-1β signal is colocalized with GFAP, and not with Iba-1 or NeuN, in the dorsal hippocampus in stressed and non-stressed animals A. Representative images of IL-1β, NeuN, Iba-1, and GFAP immunoreactivity in the dentate gyrus of the DH (AP −3.36 mm from bregma) acquired at 20X are shown. Because we did not detect any differences in colocalization between stressed and non-stressed rats, all images here are taken from animals that received stress exposure. Bitplane Imaris was used for background subtraction to better visualize individual cells presented. B. Bitplane Imaris software was used to calculate the colocalization of the IL-1β signal with GFAP, Iba-1, and NeuN. Colocalization analyses revealed that the percent of the IL-1β signal colocalized with GFAP was significantly greater than the percent of the IL-1β signal colocalized with either Iba-1 or NeuN. * p

    Journal: Brain, behavior, and immunity

    Article Title: Hippocampal interleukin-1 mediates stress-enhanced fear learning: A potential role for astrocyte-derived interleukin-1β

    doi: 10.1016/j.bbi.2017.09.016

    Figure Lengend Snippet: IL-1β signal is colocalized with GFAP, and not with Iba-1 or NeuN, in the dorsal hippocampus in stressed and non-stressed animals A. Representative images of IL-1β, NeuN, Iba-1, and GFAP immunoreactivity in the dentate gyrus of the DH (AP −3.36 mm from bregma) acquired at 20X are shown. Because we did not detect any differences in colocalization between stressed and non-stressed rats, all images here are taken from animals that received stress exposure. Bitplane Imaris was used for background subtraction to better visualize individual cells presented. B. Bitplane Imaris software was used to calculate the colocalization of the IL-1β signal with GFAP, Iba-1, and NeuN. Colocalization analyses revealed that the percent of the IL-1β signal colocalized with GFAP was significantly greater than the percent of the IL-1β signal colocalized with either Iba-1 or NeuN. * p

    Article Snippet: The following primary antibodies were used: rabbit anti-IL-1β (1:500, Abcam, Cambridge, MA, Cat# Ab9722), mouse anti-GFAP (1:1000, ThermoFisher Scientific, Waltham, MA, Cat #MS-1376P), mouse anti-NeuN-Alexa 568 (1:1000, Abcam Cambridge, MA, ), and rabbit anti-Iba-1-biotinylated (1:500, Wako, Richmond, VA, Cat#016-26461).

    Techniques: Software

    IBA1 and GFAP immunoreactivity after treatment with DA-9803 and vehicle. a – d Fluorescent images of GFAP-positive astrocytes ( a ), IBA1-positive microglia ( b ), methoxy-XO4-labeled plaques ( c ), and colocalization of GFAP, IBA1, and methoxy-XO4 ( d ) in APP/PS1 mice treated with vehicle compound. e – h Fluorescent microscope images of GFAP-positive astrocytes ( e ), IBA1-positive microglia ( f ), methoxy-XO4-labeled plaques ( g ), and colocalization of GFAP, IBA1, and methoxy-XO4 ( h ) in APP/PS1 mice treated daily with 100 mg/kg DA-9803. Scale bar, 20 μm. i Average number of astrocytes in close proximity (plaque) and away from plaques (no plaque) in a field of view across conditions. j Average number of microglia in close proximity (plaque) and away from plaques (no plaque) in a field of view across conditions. k , l High-magnification fluorescent images of GFAP-positive astrocytes treated with vehicle ( k ) and DA-9803 ( l ). m Astrocyte process length across conditions. n Astrocyte cell body diameter across conditions. o , p High magnification fluorescent images of IBA1-positive microglia treated with vehicle ( o ) and DA-9803 ( p ). q Microglial process length across conditions. r Microglial cell body diameter across conditions. Scale bar, 10 μm. Mean ± SEM. * p

    Journal: Alzheimer's Research & Therapy

    Article Title: Novel botanical drug DA-9803 prevents deficits in Alzheimer’s mouse models

    doi: 10.1186/s13195-018-0338-2

    Figure Lengend Snippet: IBA1 and GFAP immunoreactivity after treatment with DA-9803 and vehicle. a – d Fluorescent images of GFAP-positive astrocytes ( a ), IBA1-positive microglia ( b ), methoxy-XO4-labeled plaques ( c ), and colocalization of GFAP, IBA1, and methoxy-XO4 ( d ) in APP/PS1 mice treated with vehicle compound. e – h Fluorescent microscope images of GFAP-positive astrocytes ( e ), IBA1-positive microglia ( f ), methoxy-XO4-labeled plaques ( g ), and colocalization of GFAP, IBA1, and methoxy-XO4 ( h ) in APP/PS1 mice treated daily with 100 mg/kg DA-9803. Scale bar, 20 μm. i Average number of astrocytes in close proximity (plaque) and away from plaques (no plaque) in a field of view across conditions. j Average number of microglia in close proximity (plaque) and away from plaques (no plaque) in a field of view across conditions. k , l High-magnification fluorescent images of GFAP-positive astrocytes treated with vehicle ( k ) and DA-9803 ( l ). m Astrocyte process length across conditions. n Astrocyte cell body diameter across conditions. o , p High magnification fluorescent images of IBA1-positive microglia treated with vehicle ( o ) and DA-9803 ( p ). q Microglial process length across conditions. r Microglial cell body diameter across conditions. Scale bar, 10 μm. Mean ± SEM. * p

    Article Snippet: The coronal sections were then permeabilized with Triton X-100, blocked with normal goat serum (NGS), and incubated with the various primary antibodies: 6E10 (monoclonal 6E10, 1:500; Covance), IBA1 (rabbit anti-Iba1, 1:2000; Wako), glial fibrillary acidic protein (GFAP) (mouse anti-GFAP, 1:200; Thermo Scientific), and MAP2 (mouse anti-MAP2, 1:100; Abcam) for 2 hours at room temperature.

    Techniques: Labeling, Mouse Assay, Microscopy

    CAST analysis showed that agmatine treatment modulated BMP- 2/7 expressions in neurons oligodendrocytes and BMP- 4 expressions in astrocytes oligodendrocytes following SCI. (A) Stereological counting of BMP- 2 + /MAP-2 + and (B) BMP- 2 + /Olig-2 + cells in the injured spinal cord (Th 8–Th 10 segments). The number of BMP- 2 + /MAP-2 + and BMP- 2 + /Olig-2 + cells were significantly increased in the Agm treated group ( n = 5) compared with the EC group ( n = 5) at 7 35 DPI and 7 14 DPI (C) BMP- 7 + /MAP-2 + and (D) BMP- 7 + /Olig-2 + cells in the Th 8–Th 10 segments of the injured spinal cord. The number of BMP-7 + /MAP-2 + and BMP-7 + /Olig-2 + cells were significantly increased in the Agm treated group ( n = 5) compared with the EC group ( n = 5) at 7 14 days and 7 days respectively following SCI. (E) BMP- 4 + /GFAP + and (F) BMP- 4 + /Olig-2 + cells in Th 8–Th 10 segments of the injured spinal cord. The number of BMP- 4 + /GFAP + and BMP- 4 + /Olig-2 + cells were decreased at all the time periods in the Agm treated group ( n = 5) compared with the EC group ( n = 5) and the decrease was significant at 7 14 days and 14 35 days respectively following SCI. †, p

    Journal: PLoS ONE

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells

    doi: 10.1371/journal.pone.0053911

    Figure Lengend Snippet: CAST analysis showed that agmatine treatment modulated BMP- 2/7 expressions in neurons oligodendrocytes and BMP- 4 expressions in astrocytes oligodendrocytes following SCI. (A) Stereological counting of BMP- 2 + /MAP-2 + and (B) BMP- 2 + /Olig-2 + cells in the injured spinal cord (Th 8–Th 10 segments). The number of BMP- 2 + /MAP-2 + and BMP- 2 + /Olig-2 + cells were significantly increased in the Agm treated group ( n = 5) compared with the EC group ( n = 5) at 7 35 DPI and 7 14 DPI (C) BMP- 7 + /MAP-2 + and (D) BMP- 7 + /Olig-2 + cells in the Th 8–Th 10 segments of the injured spinal cord. The number of BMP-7 + /MAP-2 + and BMP-7 + /Olig-2 + cells were significantly increased in the Agm treated group ( n = 5) compared with the EC group ( n = 5) at 7 14 days and 7 days respectively following SCI. (E) BMP- 4 + /GFAP + and (F) BMP- 4 + /Olig-2 + cells in Th 8–Th 10 segments of the injured spinal cord. The number of BMP- 4 + /GFAP + and BMP- 4 + /Olig-2 + cells were decreased at all the time periods in the Agm treated group ( n = 5) compared with the EC group ( n = 5) and the decrease was significant at 7 14 days and 14 35 days respectively following SCI. †, p

    Article Snippet: The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore).

    Techniques:

    Confocal microscopic images of BMP- 2/4/7 expressions in neurons, oligodendrocytes and astrocytes after agmatine treatment following SCI. (A) Dual immunofluorescence was done to localize the BMP- 2 expression in neurons (MAP-2) and oligodendrocytes (Olig-2) after SCI. The BMP- 2 + /MAP- 2 + BMP- 2 + /Olig-2 + cells around the lesion site were higher in the Agm treated group ( n = 4) compared with the EC group ( n = 4) at7 DPI. Scale bars: 50 µm. (B) The immunostaining of BMP- 7 (red) with MAP-2 (green) showed increased number of BMP- 7 + /MAP-2 + cells in the Agm treated group ( n = 5) when compared with the EC group ( n = 5) at 7 days after SCI. The immunostaining of BMP- 7 with Olig-2 showed an increased number of BMP- 7 + /Olig-2 + cells in the Agm treated group ( n = 5) compared to EC group ( n = 5) at 14 DPI. Scale bars: 10 µm. (C) The immuno co-localization of BMP- 4 + cells in astrocytes (GFAP) showed reduced number of BMP- 4 + /GFAP + cellsaround the lesion site in the Agm treated group ( n = 4) compared with the EC group ( n = 4) at7 DPI. Whereas, BMP- 4 + /Olig-2 + cells were higher in the Agm treated group compared with EC at 35 DPI. The scale bars: 10 µm.

    Journal: PLoS ONE

    Article Title: The Multifaceted Effects of Agmatine on Functional Recovery after Spinal Cord Injury through Modulations of BMP-2/4/7 Expressions in Neurons and Glial Cells

    doi: 10.1371/journal.pone.0053911

    Figure Lengend Snippet: Confocal microscopic images of BMP- 2/4/7 expressions in neurons, oligodendrocytes and astrocytes after agmatine treatment following SCI. (A) Dual immunofluorescence was done to localize the BMP- 2 expression in neurons (MAP-2) and oligodendrocytes (Olig-2) after SCI. The BMP- 2 + /MAP- 2 + BMP- 2 + /Olig-2 + cells around the lesion site were higher in the Agm treated group ( n = 4) compared with the EC group ( n = 4) at7 DPI. Scale bars: 50 µm. (B) The immunostaining of BMP- 7 (red) with MAP-2 (green) showed increased number of BMP- 7 + /MAP-2 + cells in the Agm treated group ( n = 5) when compared with the EC group ( n = 5) at 7 days after SCI. The immunostaining of BMP- 7 with Olig-2 showed an increased number of BMP- 7 + /Olig-2 + cells in the Agm treated group ( n = 5) compared to EC group ( n = 5) at 14 DPI. Scale bars: 10 µm. (C) The immuno co-localization of BMP- 4 + cells in astrocytes (GFAP) showed reduced number of BMP- 4 + /GFAP + cellsaround the lesion site in the Agm treated group ( n = 4) compared with the EC group ( n = 4) at7 DPI. Whereas, BMP- 4 + /Olig-2 + cells were higher in the Agm treated group compared with EC at 35 DPI. The scale bars: 10 µm.

    Article Snippet: The following primary antibodies were used overnight at 4°C: anti-mouse GFAP (1∶500, monoclonal antibody; Thermo), anti-mouse BMP-2 (1∶500, monoclonal antibody; Abcam), anti-mouse BMP-4 (1∶250, polyclonal antibody; Santa-cruz), anti-rabbit BMP-7 (1∶500, monoclonal antibody; Abcam), anti-mouse MAP-2 (1∶500, monoclonal antibody; Sigma), anti-goat Olig-2 (1∶250, polyclonal antibody; Santa-cruz), anti-rat MBP (1∶500, Abcam), anti-mouse Neuronal Nuclei (NeuN) (1∶500, monoclonal antibody; Millipore), and anti-rabbit neurofilament (NF) (1∶500, monoclonal antibody; Millipore), anti-rabbit 5-HT (1∶10000, monoclonal antibody; Abcam), anti-rabbit p53 (1∶500, monoclonal antibody; santa-cruz) and anti-rabbit NG2 (1∶500, monoclonal antibody; Millipore).

    Techniques: Immunofluorescence, Expressing, Immunostaining

    Neonatal intravenous injection of AAV8.EFS.GFP enables neuronal transduction. a Brain imaged with fluorescence microscope in CD-1 pups injected intravenously with AAV8.EFS. GFP (GFP) and uninjected controls. b , c Representative images of GFP immunostaining in brain at b low and c at higher magnifications in mice injected with the GFP vector and uninjected controls show a decreasing rostro-caudal gradient with preferential transduction of forebrain and midbrain. d Computational quantification of GFP immunostaining showed a significant increase in AAV8.EFS. GFP -injected versus uninjected littermates ( n = 4). e Immunofluorescence of cortical staining for DAPI (blue), GFP (green), and NeuN, GFAP, Olig-2, CD68 (red) identifying neurons, astrocytes, oligodendrocytes and microglial cells, respectively. f Colocalisation measured by Pearson’s coefficient showed a restricted neuronal transduction. Horizontal lines display the mean ± SEM. d Unpaired two-tailed Student’s t test * p

    Journal: Nature Communications

    Article Title: Argininosuccinic aciduria fosters neuronal nitrosative stress reversed by Asl gene transfer

    doi: 10.1038/s41467-018-05972-1

    Figure Lengend Snippet: Neonatal intravenous injection of AAV8.EFS.GFP enables neuronal transduction. a Brain imaged with fluorescence microscope in CD-1 pups injected intravenously with AAV8.EFS. GFP (GFP) and uninjected controls. b , c Representative images of GFP immunostaining in brain at b low and c at higher magnifications in mice injected with the GFP vector and uninjected controls show a decreasing rostro-caudal gradient with preferential transduction of forebrain and midbrain. d Computational quantification of GFP immunostaining showed a significant increase in AAV8.EFS. GFP -injected versus uninjected littermates ( n = 4). e Immunofluorescence of cortical staining for DAPI (blue), GFP (green), and NeuN, GFAP, Olig-2, CD68 (red) identifying neurons, astrocytes, oligodendrocytes and microglial cells, respectively. f Colocalisation measured by Pearson’s coefficient showed a restricted neuronal transduction. Horizontal lines display the mean ± SEM. d Unpaired two-tailed Student’s t test * p

    Article Snippet: Antibodies used in this study include the following: rabbit polyclonal anti-GFP (1:10,000 for immunohistochemistry (IHC), 1:4000 for immunofluorescence (IF); Ab290, Abcam, Cambridge, UK), chicken polyclonal anti-GFP (1:1000 for IF; Ab13970, Abcam, Cambridge, UK), mouse polyclonal anti-nitrotyrosine (1:800 for IHC; 06-284, Merck Millipore, Temecula, CA, USA), mouse monoclonal anti-nitrotyrosine clone 1A6 (1:100 for IF and western blot (WB); 05-233, Merck Millipore, Temecula, CA, USA), rabbit polyclonal anti-nNOS (1:200 for IHC, 1:100 for IF; bs10197R, Bioss antibodies, Woburn, MA, USA), rabbit polyclonal anti-iNOS (1:500 for IHC, 1:100 for IF; NBP1-50606, Novus Biologicals, Abingdon, UK), mouse purified anti-eNOS (1:300 for IHC, 1:100 for IF; 610296, BD Transduction Lab), mouse monoclonal anti-GFAP (1:500 for IHC, 1:250 for IF; MAB3402, Merck Millipore, Temecula, CA, USA), rat monoclonal anti-GFAP (1:250 for IF, 13-0300, ThermoFisher Scientific, Rockford, IL, USA), rat monoclonal anti-CD68 (1:100 for IHC and IF; MCA1957, Bio-Rad, Oxford, UK), rabbit polyclonal anti-ASL (1:1000 for IHC; Ab97370, Abcam, Cambridge, UK), rabbit polyclonal anti-Olig-2 (1:100 for IF; Ab9610, Abcam, Cambridge, UK), mouse monoclonal anti-NeuN (1:1000 for IF; Millipore, Billerica, MA, USA), rabbit monoclonal anti-NeuN (1:1000 for IF; , Abcam, Cambridge, UK), mouse monoclonal anti-CD31 clone 390 (1:20 in IF, ThermoFisher Scientific, Rockford, IL, USA) and goat anti-rabbit secondary antibody (1:1000 for IHC and WB; Vector, Burlingame, CA, USA).

    Techniques: Injection, Transduction, Fluorescence, Microscopy, Immunostaining, Mouse Assay, Plasmid Preparation, Immunofluorescence, Staining, Two Tailed Test