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  • 94
    Millipore antibodies against blbp
    Additional control data for Pcdhα function in cortical neuron migration. ( A ) RT-PCR of members of the Pcdhα family in E9, E10, and E14 embryonic mouse brain. ( B ) Western blot and its quantification of lysates of 293 T cells transfected with control or αKD plasmids. n = 3 experiments for each group. Statistical significance was assessed using one-way ANOVA, followed by a post hoc Tukey’s multiple comparisons test. ( C ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with a BrdU antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5, injected with BrdU at E16.5, and sectioned at E17.5. ( D ) Percentage of <t>GFP</t> + and BrdU + cells among GFP + cells in the VZ/SVZ region shown in ( C ). n = 5 brains for each group. Student’s t test. ( E ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with Tbr2 antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5. ( F ) Percentage of GFP + and BrdU + intermediate progenitor cells (IPCs) in GFP + cells in the VZ/SVZ region shown in ( E ). n = 3 brains for each group. Student’s t test. ( G ) The CP region of E17.5 cortical coronal sections immunostained with an anti-brain lipid binding protein <t>(BLBP)</t> antibody of embryonic brains electroporated at E15.5. Arrows, BLBP-labeled radial glia cells. ( H ) E19.5 cortical coronal sections immunostaining for activated Caspase3 of embryonic brains electroporated at E15.5. Nuclei were counterstained with DAPI. ( I ) Cortical coronal sections of E19.5 Pcdhα knockout mouse brains which were in utero electroporated at E15.5 with GFP plasmids. Slides were counterstained with DAPI. Quantification of GFP + cell distribution across the cortex is shown on the right. n = 6 brains for each group. Data as mean ±SEM. **p
    Antibodies Against Blbp, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti blbp
    Nkx6.1 expression in ventral spinal cord astrocytes. A-E : Transverse sections from wild-type mice spinal cord at early postnatal stages were double immunostained with anti-Nkx6.1 in conjunction with <t>anti-Olig2,</t> anti-Sox10, <t>anti-BLBP,</t> anti-GFAP, or anti-GS. Double-positive cells are represented by arrows. Orthogonal reconstructions of confocal images at z-axis level are shown in side panels (along the right-hand edge and beneath). Note that all Nkx6.1+ cells in ventral white matter (WM) co-express GFAP. F-G : Spinal cord sections from P4 wild-type animal were subjected to Fgfr3 (blue) ISH followed by anti-Nkx6.1 (brown) immunostaining. Representative double positive cells in WM and gray matter (GM) are indicated by red arrows. The insets are the higher magnification of double positive cells. H-I : Transverse sections from wild-type mice spinal cord at P0 were double immunostained with anti-Nkx6.1 and anti-Iba1 or anti-NeuN antibodies. Arrows represent double-positive cells in GM. Scale bars 50 µm.
    Anti Blbp, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 166 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    blbp  (Abcam)
    95
    Abcam blbp
    Reduced neocortical astrogliogenesis in Yap <t>nestin</t> -CKO mice. (A) Double immunostaining of <t>BLBP</t> (green) and NeuN (red) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice (sagittal sections). (B) Quantitative analysis of BLBP ( n =18 for neocortex per group, and n =5 for hippocampus per group) fluorescence intensity as shown in A. (C) Western blot analysis of BLBP expression in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (D) Quantitative analysis of western blot data as shown in C ( n =3 per group, normalized to WT). (E) Immunostaining of aldolase C (green) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (F,G) Quantitative analysis of (F) the density of aldolase C + cells and (G) the relative aldolase C fluorescence intensity ( n =8 per group, normalized to WT) as shown in E. Selected regions in Yap f/f (a,b) and Yap nestin -CKO (c,d) mice are shown at higher magnification. Data are mean±s.e.m. ** P
    Blbp, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Blbp, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 372 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Blbp, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Santa Cruz Biotechnology anti blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Anti Blbp, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam rabbit polyclonal anti blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Rabbit Polyclonal Anti Blbp, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Blbp, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Carl Zeiss blbp
    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker <t>BLBP</t> (A’, red). Merged images (A ″ ). Immuno-staining for <t>GFAP</t> (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).
    Blbp, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit anti blbp
    Comparative analysis of reprogramming potentials of distinct factor combinations. (A) The schematic illustration depicting the strategy for comparing the reprogramming efficiency of distinct factor combinations. (B) Time-course immunofluorescence analysis for comparing reprogramming potentials of distinct factor combinations. Scale bars, 100 μ m. (C) Morphology of hiNSC clusters at 2 weeks after transduction. Scale bars, 100 μ m. (D) The number of <t>BLBP</t> + / <t>MSI1</t> + colonies were counted in a time-course manner. Data are presented as mean±SD from six independent experiments. *p
    Rabbit Anti Blbp, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Millipore mouse anti blbp antibody
    Genome wide correlation analysis between Ca 2+ drug sensitivity and gene expression. (A) Nine novel GIC lines were subjected to Thapsigargin dose response analysis (0.1–100 µM), showing different response to moderate drug doses. (B, C) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and (B) NES and (C) <t>FABP7/BLBP</t> mRNA expression. U3047-MG was considered an outlier in the NES graph (marked in red) and excluded form the analysis. (D) Western blot analysis showing BLBP (FABP7) protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines, with β-actin as loading control. (E) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and <t>GRIA1</t> mRNA expression. (F) Western blot analysis showing GRIA1 protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines. β-actin was used as loading control.
    Mouse Anti Blbp Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck KGaA anti blbp
    Genome wide correlation analysis between Ca 2+ drug sensitivity and gene expression. (A) Nine novel GIC lines were subjected to Thapsigargin dose response analysis (0.1–100 µM), showing different response to moderate drug doses. (B, C) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and (B) NES and (C) <t>FABP7/BLBP</t> mRNA expression. U3047-MG was considered an outlier in the NES graph (marked in red) and excluded form the analysis. (D) Western blot analysis showing BLBP (FABP7) protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines, with β-actin as loading control. (E) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and <t>GRIA1</t> mRNA expression. (F) Western blot analysis showing GRIA1 protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines. β-actin was used as loading control.
    Anti Blbp, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Follett blbp antibody
    Genome wide correlation analysis between Ca 2+ drug sensitivity and gene expression. (A) Nine novel GIC lines were subjected to Thapsigargin dose response analysis (0.1–100 µM), showing different response to moderate drug doses. (B, C) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and (B) NES and (C) <t>FABP7/BLBP</t> mRNA expression. U3047-MG was considered an outlier in the NES graph (marked in red) and excluded form the analysis. (D) Western blot analysis showing BLBP (FABP7) protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines, with β-actin as loading control. (E) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and <t>GRIA1</t> mRNA expression. (F) Western blot analysis showing GRIA1 protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines. β-actin was used as loading control.
    Blbp Antibody, supplied by Follett, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Additional control data for Pcdhα function in cortical neuron migration. ( A ) RT-PCR of members of the Pcdhα family in E9, E10, and E14 embryonic mouse brain. ( B ) Western blot and its quantification of lysates of 293 T cells transfected with control or αKD plasmids. n = 3 experiments for each group. Statistical significance was assessed using one-way ANOVA, followed by a post hoc Tukey’s multiple comparisons test. ( C ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with a BrdU antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5, injected with BrdU at E16.5, and sectioned at E17.5. ( D ) Percentage of GFP + and BrdU + cells among GFP + cells in the VZ/SVZ region shown in ( C ). n = 5 brains for each group. Student’s t test. ( E ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with Tbr2 antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5. ( F ) Percentage of GFP + and BrdU + intermediate progenitor cells (IPCs) in GFP + cells in the VZ/SVZ region shown in ( E ). n = 3 brains for each group. Student’s t test. ( G ) The CP region of E17.5 cortical coronal sections immunostained with an anti-brain lipid binding protein (BLBP) antibody of embryonic brains electroporated at E15.5. Arrows, BLBP-labeled radial glia cells. ( H ) E19.5 cortical coronal sections immunostaining for activated Caspase3 of embryonic brains electroporated at E15.5. Nuclei were counterstained with DAPI. ( I ) Cortical coronal sections of E19.5 Pcdhα knockout mouse brains which were in utero electroporated at E15.5 with GFP plasmids. Slides were counterstained with DAPI. Quantification of GFP + cell distribution across the cortex is shown on the right. n = 6 brains for each group. Data as mean ±SEM. **p

    Journal: eLife

    Article Title: Alpha protocadherins and Pyk2 kinase regulate cortical neuron migration and cytoskeletal dynamics via Rac1 GTPase and WAVE complex in mice

    doi: 10.7554/eLife.35242

    Figure Lengend Snippet: Additional control data for Pcdhα function in cortical neuron migration. ( A ) RT-PCR of members of the Pcdhα family in E9, E10, and E14 embryonic mouse brain. ( B ) Western blot and its quantification of lysates of 293 T cells transfected with control or αKD plasmids. n = 3 experiments for each group. Statistical significance was assessed using one-way ANOVA, followed by a post hoc Tukey’s multiple comparisons test. ( C ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with a BrdU antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5, injected with BrdU at E16.5, and sectioned at E17.5. ( D ) Percentage of GFP + and BrdU + cells among GFP + cells in the VZ/SVZ region shown in ( C ). n = 5 brains for each group. Student’s t test. ( E ) The VZ/SVZ region of E17.5 cortical coronal sections immunostained with Tbr2 antibody. Embryonic mice were electroporated with control or αKD plasmids at E15.5. ( F ) Percentage of GFP + and BrdU + intermediate progenitor cells (IPCs) in GFP + cells in the VZ/SVZ region shown in ( E ). n = 3 brains for each group. Student’s t test. ( G ) The CP region of E17.5 cortical coronal sections immunostained with an anti-brain lipid binding protein (BLBP) antibody of embryonic brains electroporated at E15.5. Arrows, BLBP-labeled radial glia cells. ( H ) E19.5 cortical coronal sections immunostaining for activated Caspase3 of embryonic brains electroporated at E15.5. Nuclei were counterstained with DAPI. ( I ) Cortical coronal sections of E19.5 Pcdhα knockout mouse brains which were in utero electroporated at E15.5 with GFP plasmids. Slides were counterstained with DAPI. Quantification of GFP + cell distribution across the cortex is shown on the right. n = 6 brains for each group. Data as mean ±SEM. **p

    Article Snippet: The following antibodies were used for immunocytochemistry and immunohistochemistry: mouse anti-Tuj1 (1:300, Covance), rabbit anti-αCD (1:500, Synaptic Systems), rabbit anti-GFP (1:1000, Invitrogen), rabbit anti-BLBP (Brain lipid binding protein) (1:500, Chemicon), mouse anti-GM130 (1:1000, BD Bioscience), rat anti-BrdU (1:1000, Bio-Rad), rabbit anti-activated caspase 3 (1:500; Cell Signaling Technology), rabbit anti-Tbr2 (1:500, Abcam), rabbit anti-WAVE2 (1:500, Millipore), goat anti-rabbit Alexa Fluor 488 (1:300, Molecular Probes), goat anti-rabbit Alexa Fluor 568 (1:300, Molecular Probes), goat anti-mouse Alexa Fluor 568 (1:300, Molecular Probes), goat anti-mouse Alexa Fluor 647 (1:300, Molecular Probes).

    Techniques: Migration, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Mouse Assay, Injection, Binding Assay, Labeling, Immunostaining, Knock-Out, In Utero

    Nkx6.1 expression in ventral spinal cord astrocytes. A-E : Transverse sections from wild-type mice spinal cord at early postnatal stages were double immunostained with anti-Nkx6.1 in conjunction with anti-Olig2, anti-Sox10, anti-BLBP, anti-GFAP, or anti-GS. Double-positive cells are represented by arrows. Orthogonal reconstructions of confocal images at z-axis level are shown in side panels (along the right-hand edge and beneath). Note that all Nkx6.1+ cells in ventral white matter (WM) co-express GFAP. F-G : Spinal cord sections from P4 wild-type animal were subjected to Fgfr3 (blue) ISH followed by anti-Nkx6.1 (brown) immunostaining. Representative double positive cells in WM and gray matter (GM) are indicated by red arrows. The insets are the higher magnification of double positive cells. H-I : Transverse sections from wild-type mice spinal cord at P0 were double immunostained with anti-Nkx6.1 and anti-Iba1 or anti-NeuN antibodies. Arrows represent double-positive cells in GM. Scale bars 50 µm.

    Journal: PLoS ONE

    Article Title: Control of Astrocyte Progenitor Specification, Migration and Maturation by Nkx6.1 Homeodomain Transcription Factor

    doi: 10.1371/journal.pone.0109171

    Figure Lengend Snippet: Nkx6.1 expression in ventral spinal cord astrocytes. A-E : Transverse sections from wild-type mice spinal cord at early postnatal stages were double immunostained with anti-Nkx6.1 in conjunction with anti-Olig2, anti-Sox10, anti-BLBP, anti-GFAP, or anti-GS. Double-positive cells are represented by arrows. Orthogonal reconstructions of confocal images at z-axis level are shown in side panels (along the right-hand edge and beneath). Note that all Nkx6.1+ cells in ventral white matter (WM) co-express GFAP. F-G : Spinal cord sections from P4 wild-type animal were subjected to Fgfr3 (blue) ISH followed by anti-Nkx6.1 (brown) immunostaining. Representative double positive cells in WM and gray matter (GM) are indicated by red arrows. The insets are the higher magnification of double positive cells. H-I : Transverse sections from wild-type mice spinal cord at P0 were double immunostained with anti-Nkx6.1 and anti-Iba1 or anti-NeuN antibodies. Arrows represent double-positive cells in GM. Scale bars 50 µm.

    Article Snippet: The dilution ratio of antibodies is as follows: anti-S100B (Millipore Bioscience Research Reagents, 1∶1000), anti-GS (Sigma, 1∶500), anti-BLBP (Abcam, 1∶500), anti-Olig2 (Abcam, 1∶3000), anti-NeuN (Abcam, 1∶100), anti-Iba1 (Wako, 1∶500), anti-Sox10 (1∶3000, kind gift of Dr. Michael Wegner), anti-GFAP (Millipore Bioscience Research Reagents, 1∶500), anti-Nkx6.1 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA; 1∶50) .

    Techniques: Expressing, Mouse Assay, In Situ Hybridization, Immunostaining

    Oligodendrocyte progenitors derived from cerebral organoids. (A) Schematic representation of OPC isolation and culture. (B) Phase contrast of immature oligodendrocytes with initial ramifications (arrows in white). OPCs were stained for (C,D) olig2, oligodendrocyte lineage transcription factor, (E–G) BLBP and PAX6 progenitor markers, (H) CNPase (CNP), enzyme of early oligodendrocyte formation and myelin precursor, and (I) PDGFRA oligodendrocyte lineage marker. Scale bars shown are (B) 400 μm, (D,G) 30 μm and (H,I) 50 μm.

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: Human Cerebral Organoids and Fetal Brain Tissue Share Proteomic Similarities

    doi: 10.3389/fcell.2019.00303

    Figure Lengend Snippet: Oligodendrocyte progenitors derived from cerebral organoids. (A) Schematic representation of OPC isolation and culture. (B) Phase contrast of immature oligodendrocytes with initial ramifications (arrows in white). OPCs were stained for (C,D) olig2, oligodendrocyte lineage transcription factor, (E–G) BLBP and PAX6 progenitor markers, (H) CNPase (CNP), enzyme of early oligodendrocyte formation and myelin precursor, and (I) PDGFRA oligodendrocyte lineage marker. Scale bars shown are (B) 400 μm, (D,G) 30 μm and (H,I) 50 μm.

    Article Snippet: For isolated and differentiated OPCs, primary antibodies were: anti-BLBP (1:200, AB110099, Abcam), anti-CNPase (1:200, #5664, Cell Signaling), anti-PDGFRA (1:1000, #3174, Cell Signaling), anti-olig2 (1:100, MABN50, Millipore), and anti-PAX6 (1:200, sc-11357, KloneLife).

    Techniques: Derivative Assay, Isolation, Staining, Marker

    GLI3R rescues delayed cell differentiation in SuFu -deficient cerebella. A–I , Calbindin staining (green) reveals a complete restoration of normal Purkinje cell morphology in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella by P14 ( C , F ). Hoxb.7-Cre;SuFu −/ loxP Purkinje cells show decreased calbindin expression and are abnormal in dendritic morphology at P7 and P14 ( B , E ). By P21, Purkinje cells in Hoxb.7-Cre;SuFu −/ loxP cerebella are comparable to those observed in control and Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( G–I ). GFAP expression (red), a marker of Bergmann glia, in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella is comparable to that observed in control cerebella as early as P7 ( C , F , I ). In Hoxb.7-Cre;SuFu −/ loxP mutants, Bergmann glia are severely disorganized and radial fibers are misaligned ( B , E , H ). J–L , BLBP-positive Bergmann glia cell somas (white arrowheads) are severely disorganized and fail to alight with Purkinje cells (yellow asterisks) in Hoxb.7-Cre;SuFu −/ loxP cerebella. Normal Bergmann glia organization is restored with the exception of some Bergmann glia cell somas that are detected in the inner granule layer (IGL) in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella. M–O , PAX6 and NeuN double staining demonstrates normal expression of the granule cell markers in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( L ). Note that PAX6 expression is slightly upregulated in the IGL of Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella. In Hoxb.7-Cre;SuFu −/ loxP cerebella, the proportion of PAX6-expressing cells in the presumptive EGL to NeuN-expressing cells in the IGL is significantly increased compared with control and Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( K ). ML, Molecular layer. Scale bars: (in G ) A–I , 200 μm; J–L , 100 μm; (in M ) M–O as supplemental material).

    Journal: The Journal of Neuroscience

    Article Title: Suppressor of Fused Controls Mid-Hindbrain Patterning and Cerebellar Morphogenesis via GLI3 Repressor

    doi: 10.1523/JNEUROSCI.2166-10.2011

    Figure Lengend Snippet: GLI3R rescues delayed cell differentiation in SuFu -deficient cerebella. A–I , Calbindin staining (green) reveals a complete restoration of normal Purkinje cell morphology in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella by P14 ( C , F ). Hoxb.7-Cre;SuFu −/ loxP Purkinje cells show decreased calbindin expression and are abnormal in dendritic morphology at P7 and P14 ( B , E ). By P21, Purkinje cells in Hoxb.7-Cre;SuFu −/ loxP cerebella are comparable to those observed in control and Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( G–I ). GFAP expression (red), a marker of Bergmann glia, in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella is comparable to that observed in control cerebella as early as P7 ( C , F , I ). In Hoxb.7-Cre;SuFu −/ loxP mutants, Bergmann glia are severely disorganized and radial fibers are misaligned ( B , E , H ). J–L , BLBP-positive Bergmann glia cell somas (white arrowheads) are severely disorganized and fail to alight with Purkinje cells (yellow asterisks) in Hoxb.7-Cre;SuFu −/ loxP cerebella. Normal Bergmann glia organization is restored with the exception of some Bergmann glia cell somas that are detected in the inner granule layer (IGL) in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella. M–O , PAX6 and NeuN double staining demonstrates normal expression of the granule cell markers in Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( L ). Note that PAX6 expression is slightly upregulated in the IGL of Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella. In Hoxb.7-Cre;SuFu −/ loxP cerebella, the proportion of PAX6-expressing cells in the presumptive EGL to NeuN-expressing cells in the IGL is significantly increased compared with control and Hoxb.7-Cre;SuFu −/ loxP ; Gli3 Δ 699 cerebella ( K ). ML, Molecular layer. Scale bars: (in G ) A–I , 200 μm; J–L , 100 μm; (in M ) M–O as supplemental material).

    Article Snippet: Immunofluorescence was performed on PFA- or formalin-fixed, paraffin-embedded tissue sections using the following primary antibodies: mouse anti-calbindin D28K (1:200, Sigma), rabbit anti-SUFU (1:1500, kindly provided by C.-C. Hui), rabbit anti-BLBP (brain lipid-binding protein) (1:200, Abcam), mouse anti-GFAP (1:200, Millipore), mouse anti-NeuN (1:100, Millipore), rabbit anti-Pax2 (1:200, Covance), rabbit anti-Pax6 (1:300, Covance), mouse anti-PCNA (proliferating cell nuclear antigen) (1:500, Cell Signaling Technology), rabbit anti-Nestin (1:500, Abcam), mouse anti-Ki67 (1:200, BD Biosciences), and mouse anti-βIII tubulin (1:400, Covance).

    Techniques: Cell Differentiation, Staining, Expressing, Marker, Double Staining

    Hoxb.7-Cre is expressed in the mid-hindbrain and deletes SuFu in the cerebellum. A , B , Hoxb.7-Cre mice were crossed to ROSA26 mice to examine Cre expression. Hoxb.7 activates Cre in the mid-hindbrain at E9.5, before the onset of cerebellar morphogenesis ( A ). β-Galactosidase activity indicates Cre expression in the mid-hindbrain (arrows) and the Wolffian duct ( B , arrowhead). C , Strong Cre activity is detected in the cerebellum at E18.5. LacZ stain is denoted by blue stain and neutral red stain is denoted by pink stain. D – D‴ , SUFU and Nestin, a specific marker of radial glia precursors, are expressed in an overlapping pattern in the ventricular zone of E13.5 wild-type cerebella (arrows, D″ , D‴ ). E – E‴ , At E18.5, SUFU is detected in the Purkinje cell layer (PCL) and is expressed in a pattern similar to that of Bergmann glia radial fibers, identified by BLBP expression ( E″ , E‴ , arrows). F , F′ , SUFU is also observed in the presumptive deep cerebellar nuclei (DCN). G , G′ , The inner external granule layer (iEGL) and PCL express SUFU, whereas the outer EGL (oEGL) lacks SUFU expression at P6. H , H′ , SUFU is specifically expressed in Purkinje cells at P21. I , Western blot analysis reveals a marked decrease in SUFU protein levels in P7 cerebellar protein lysates. 1, SuFu +/ loxP ; 2–4, SuFu −/ loxP ; 5–7, Hoxb.7-Cre;SuFu −/ loxP ; 8, SuFu −/− . Scale bars: (in C ) A , B , 1.1 mm; C , 200 μm; (in D ) D–H , 200 μm; D′–H′ , 100 μm; (in D″ ) D″ , D‴ , E″ , E‴ , 30 μm.

    Journal: The Journal of Neuroscience

    Article Title: Suppressor of Fused Controls Mid-Hindbrain Patterning and Cerebellar Morphogenesis via GLI3 Repressor

    doi: 10.1523/JNEUROSCI.2166-10.2011

    Figure Lengend Snippet: Hoxb.7-Cre is expressed in the mid-hindbrain and deletes SuFu in the cerebellum. A , B , Hoxb.7-Cre mice were crossed to ROSA26 mice to examine Cre expression. Hoxb.7 activates Cre in the mid-hindbrain at E9.5, before the onset of cerebellar morphogenesis ( A ). β-Galactosidase activity indicates Cre expression in the mid-hindbrain (arrows) and the Wolffian duct ( B , arrowhead). C , Strong Cre activity is detected in the cerebellum at E18.5. LacZ stain is denoted by blue stain and neutral red stain is denoted by pink stain. D – D‴ , SUFU and Nestin, a specific marker of radial glia precursors, are expressed in an overlapping pattern in the ventricular zone of E13.5 wild-type cerebella (arrows, D″ , D‴ ). E – E‴ , At E18.5, SUFU is detected in the Purkinje cell layer (PCL) and is expressed in a pattern similar to that of Bergmann glia radial fibers, identified by BLBP expression ( E″ , E‴ , arrows). F , F′ , SUFU is also observed in the presumptive deep cerebellar nuclei (DCN). G , G′ , The inner external granule layer (iEGL) and PCL express SUFU, whereas the outer EGL (oEGL) lacks SUFU expression at P6. H , H′ , SUFU is specifically expressed in Purkinje cells at P21. I , Western blot analysis reveals a marked decrease in SUFU protein levels in P7 cerebellar protein lysates. 1, SuFu +/ loxP ; 2–4, SuFu −/ loxP ; 5–7, Hoxb.7-Cre;SuFu −/ loxP ; 8, SuFu −/− . Scale bars: (in C ) A , B , 1.1 mm; C , 200 μm; (in D ) D–H , 200 μm; D′–H′ , 100 μm; (in D″ ) D″ , D‴ , E″ , E‴ , 30 μm.

    Article Snippet: Immunofluorescence was performed on PFA- or formalin-fixed, paraffin-embedded tissue sections using the following primary antibodies: mouse anti-calbindin D28K (1:200, Sigma), rabbit anti-SUFU (1:1500, kindly provided by C.-C. Hui), rabbit anti-BLBP (brain lipid-binding protein) (1:200, Abcam), mouse anti-GFAP (1:200, Millipore), mouse anti-NeuN (1:100, Millipore), rabbit anti-Pax2 (1:200, Covance), rabbit anti-Pax6 (1:300, Covance), mouse anti-PCNA (proliferating cell nuclear antigen) (1:500, Cell Signaling Technology), rabbit anti-Nestin (1:500, Abcam), mouse anti-Ki67 (1:200, BD Biosciences), and mouse anti-βIII tubulin (1:400, Covance).

    Techniques: Mouse Assay, Expressing, Activity Assay, Staining, Marker, Western Blot

    Cerebellar cell differentiation and migration is abnormal in the absence of SuFu . A–F , PAX6 staining reveals that Hoxb.7-Cre;SuFu −/ loxP granule cell precursors are delayed in cell specification and form a defective EGL. G–L , As marked by BLBP, Bergmann glia arise from the ventricular zone (VZ) late and display abnormal migration and disorganization in Hoxb.7-Cre;SuFu −/ loxP mutants. M–R , Calbindin-expressing Purkinje cell precursors in Hoxb.7-Cre;SuFu −/ loxP cerebella are delayed in specification and fail to form a thin layer underneath the granule cells. Note that the number of Purkinje cells is markedly reduced compared with that of control cerebella. S–X , PAX2 staining demonstrates that GABAergic interneuron precursors in mutant cerebella exhibit a delay in cell specification and are significantly increased in number compared with control cerebella. Also, ectopic localization of interneuron precursors is detected in the isthmus (IS) at E18.5. CB, cerebellum; PCL, Purkinje cell layer; RL, rhombic lip. Arrowheads indicate cell marker-positive precursor cells. Scale bar: (in D ) A–F , M–X , 200 μm; G–L as supplemental material).

    Journal: The Journal of Neuroscience

    Article Title: Suppressor of Fused Controls Mid-Hindbrain Patterning and Cerebellar Morphogenesis via GLI3 Repressor

    doi: 10.1523/JNEUROSCI.2166-10.2011

    Figure Lengend Snippet: Cerebellar cell differentiation and migration is abnormal in the absence of SuFu . A–F , PAX6 staining reveals that Hoxb.7-Cre;SuFu −/ loxP granule cell precursors are delayed in cell specification and form a defective EGL. G–L , As marked by BLBP, Bergmann glia arise from the ventricular zone (VZ) late and display abnormal migration and disorganization in Hoxb.7-Cre;SuFu −/ loxP mutants. M–R , Calbindin-expressing Purkinje cell precursors in Hoxb.7-Cre;SuFu −/ loxP cerebella are delayed in specification and fail to form a thin layer underneath the granule cells. Note that the number of Purkinje cells is markedly reduced compared with that of control cerebella. S–X , PAX2 staining demonstrates that GABAergic interneuron precursors in mutant cerebella exhibit a delay in cell specification and are significantly increased in number compared with control cerebella. Also, ectopic localization of interneuron precursors is detected in the isthmus (IS) at E18.5. CB, cerebellum; PCL, Purkinje cell layer; RL, rhombic lip. Arrowheads indicate cell marker-positive precursor cells. Scale bar: (in D ) A–F , M–X , 200 μm; G–L as supplemental material).

    Article Snippet: Immunofluorescence was performed on PFA- or formalin-fixed, paraffin-embedded tissue sections using the following primary antibodies: mouse anti-calbindin D28K (1:200, Sigma), rabbit anti-SUFU (1:1500, kindly provided by C.-C. Hui), rabbit anti-BLBP (brain lipid-binding protein) (1:200, Abcam), mouse anti-GFAP (1:200, Millipore), mouse anti-NeuN (1:100, Millipore), rabbit anti-Pax2 (1:200, Covance), rabbit anti-Pax6 (1:300, Covance), mouse anti-PCNA (proliferating cell nuclear antigen) (1:500, Cell Signaling Technology), rabbit anti-Nestin (1:500, Abcam), mouse anti-Ki67 (1:200, BD Biosciences), and mouse anti-βIII tubulin (1:400, Covance).

    Techniques: Cell Differentiation, Migration, Staining, Expressing, Mutagenesis, Marker

    SuFu-deficient cells undergo enhanced gliogenesis and are delayed in neuronal differentiation. A , B , Double staining of BLBP (green) and βIII tubulin (red) demonstrates that the proportion of BLBP-expressing cells versus βIII tubulin cells is significantly increased in Hoxb.7-Cre;SuFu −/ loxP cerebellar cultures (control: 0.53 ± 0.08; mutant: 3.00 ± 0.45, p

    Journal: The Journal of Neuroscience

    Article Title: Suppressor of Fused Controls Mid-Hindbrain Patterning and Cerebellar Morphogenesis via GLI3 Repressor

    doi: 10.1523/JNEUROSCI.2166-10.2011

    Figure Lengend Snippet: SuFu-deficient cells undergo enhanced gliogenesis and are delayed in neuronal differentiation. A , B , Double staining of BLBP (green) and βIII tubulin (red) demonstrates that the proportion of BLBP-expressing cells versus βIII tubulin cells is significantly increased in Hoxb.7-Cre;SuFu −/ loxP cerebellar cultures (control: 0.53 ± 0.08; mutant: 3.00 ± 0.45, p

    Article Snippet: Immunofluorescence was performed on PFA- or formalin-fixed, paraffin-embedded tissue sections using the following primary antibodies: mouse anti-calbindin D28K (1:200, Sigma), rabbit anti-SUFU (1:1500, kindly provided by C.-C. Hui), rabbit anti-BLBP (brain lipid-binding protein) (1:200, Abcam), mouse anti-GFAP (1:200, Millipore), mouse anti-NeuN (1:100, Millipore), rabbit anti-Pax2 (1:200, Covance), rabbit anti-Pax6 (1:300, Covance), mouse anti-PCNA (proliferating cell nuclear antigen) (1:500, Cell Signaling Technology), rabbit anti-Nestin (1:500, Abcam), mouse anti-Ki67 (1:200, BD Biosciences), and mouse anti-βIII tubulin (1:400, Covance).

    Techniques: Double Staining, Expressing, Mutagenesis

    Reduced neocortical astrogliogenesis in Yap nestin -CKO mice. (A) Double immunostaining of BLBP (green) and NeuN (red) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice (sagittal sections). (B) Quantitative analysis of BLBP ( n =18 for neocortex per group, and n =5 for hippocampus per group) fluorescence intensity as shown in A. (C) Western blot analysis of BLBP expression in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (D) Quantitative analysis of western blot data as shown in C ( n =3 per group, normalized to WT). (E) Immunostaining of aldolase C (green) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (F,G) Quantitative analysis of (F) the density of aldolase C + cells and (G) the relative aldolase C fluorescence intensity ( n =8 per group, normalized to WT) as shown in E. Selected regions in Yap f/f (a,b) and Yap nestin -CKO (c,d) mice are shown at higher magnification. Data are mean±s.e.m. ** P

    Journal: Development (Cambridge, England)

    Article Title: YAP stabilizes SMAD1 and promotes BMP2-induced neocortical astrocytic differentiation

    doi: 10.1242/dev.130658

    Figure Lengend Snippet: Reduced neocortical astrogliogenesis in Yap nestin -CKO mice. (A) Double immunostaining of BLBP (green) and NeuN (red) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice (sagittal sections). (B) Quantitative analysis of BLBP ( n =18 for neocortex per group, and n =5 for hippocampus per group) fluorescence intensity as shown in A. (C) Western blot analysis of BLBP expression in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (D) Quantitative analysis of western blot data as shown in C ( n =3 per group, normalized to WT). (E) Immunostaining of aldolase C (green) in neocortex and hippocampus of P1 Yap f/f and Yap nestin -CKO mice. (F,G) Quantitative analysis of (F) the density of aldolase C + cells and (G) the relative aldolase C fluorescence intensity ( n =8 per group, normalized to WT) as shown in E. Selected regions in Yap f/f (a,b) and Yap nestin -CKO (c,d) mice are shown at higher magnification. Data are mean±s.e.m. ** P

    Article Snippet: Primary antibodies were rabbit polyclonal antibodies against nestin (1:200, SAB4200394, Sigma), BLBP (1:300, Ab32423, Abcam), Ki67 (1:200, AB9260, Millipore), PH3 (1:200, 06-570, Millipore), GFAP (1:500, AB5804, Millipore), GLAST (SLC1A3 or EAAT1; 1:200, AB416, Abcam), pSMAD1/5/8 (1:200, #13820, CST), Flag (1:2000, #14793, CST), YAP (1:200, #8418/D24E4, CST), CUX1 (1:200, M222, Santa Cruz), c leaved caspase 3 (1:100, #9661, CST), TBR1 (1:200, AB10554, Millipore) or TBR2 (1:400, AB2283, Millipore), or with monoclonal antibodies against YAP (1:200, WH0010413M1, Sigma), GFAP (1:500, MAB360, Millipore), NeuN (1:500, MAB377, Millipore), MAP2 (1:500, AB5622, Millipore), Tuj1 (1:500, T8660, Sigma) and nestin (1:1000, MAB353, Sigma), or with goat polyclonal antibodies against aldolase C (1:500, SC-12065, Santa Cruz).

    Techniques: Mouse Assay, Double Immunostaining, Fluorescence, Western Blot, Expressing, Immunostaining

    Reduced local proliferation of neocortical astrocytes in Yap nestin -CKO mice. (A,B) Double immunostaining analysis of (A) Ki67 (green) and GFAP (red) or (B) PH3 (green) and GFAP (red) in cultured astrocytes from P1 Yap f/f and Yap nestin -CKO mice. (C,D) Quantitative analysis of the percentage of Ki67 + (C, n =16 per group) and PH3 + (D, n =12 per group) astrocytes among total astrocytes. (E) Scheme of BrdU incorporation experiments in F. AS, astrocyte. (F) Double immunostaining analysis of Ki67 (green) and BrdU (red) in neocortex of P0 Yap f/f and Yap nestin -CKO mice (sagittal sections). (G,H) Quantitative analysis of (G) Ki67 + or (H) BrdU + cell density in neocortex of P0 Yap f/f and Yap nestin -CKO mice ( n =4 per group) as shown in F. (I) Scheme of BrdU incorporation experiments in J. (J) Double immunostaining analysis of BLBP (green) and BrdU (red) in neocortex of P1 Yap f/f and Yap nestin -CKO mice (sagittal sections). (K,L) Quantitative analysis of (K) BrdU + cell density and (L) the percentage of BLBP + and BrdU + cells among total BLBP + cells ( n =10 per group) in neocortex of P1 Yap f/f and Yap nestin -CKO mice as shown in J. Boxed regions are shown at higher magnification. DAPI (blue) was used to stain nuclei. Data are mean±s.e.m. ** P

    Journal: Development (Cambridge, England)

    Article Title: YAP stabilizes SMAD1 and promotes BMP2-induced neocortical astrocytic differentiation

    doi: 10.1242/dev.130658

    Figure Lengend Snippet: Reduced local proliferation of neocortical astrocytes in Yap nestin -CKO mice. (A,B) Double immunostaining analysis of (A) Ki67 (green) and GFAP (red) or (B) PH3 (green) and GFAP (red) in cultured astrocytes from P1 Yap f/f and Yap nestin -CKO mice. (C,D) Quantitative analysis of the percentage of Ki67 + (C, n =16 per group) and PH3 + (D, n =12 per group) astrocytes among total astrocytes. (E) Scheme of BrdU incorporation experiments in F. AS, astrocyte. (F) Double immunostaining analysis of Ki67 (green) and BrdU (red) in neocortex of P0 Yap f/f and Yap nestin -CKO mice (sagittal sections). (G,H) Quantitative analysis of (G) Ki67 + or (H) BrdU + cell density in neocortex of P0 Yap f/f and Yap nestin -CKO mice ( n =4 per group) as shown in F. (I) Scheme of BrdU incorporation experiments in J. (J) Double immunostaining analysis of BLBP (green) and BrdU (red) in neocortex of P1 Yap f/f and Yap nestin -CKO mice (sagittal sections). (K,L) Quantitative analysis of (K) BrdU + cell density and (L) the percentage of BLBP + and BrdU + cells among total BLBP + cells ( n =10 per group) in neocortex of P1 Yap f/f and Yap nestin -CKO mice as shown in J. Boxed regions are shown at higher magnification. DAPI (blue) was used to stain nuclei. Data are mean±s.e.m. ** P

    Article Snippet: Primary antibodies were rabbit polyclonal antibodies against nestin (1:200, SAB4200394, Sigma), BLBP (1:300, Ab32423, Abcam), Ki67 (1:200, AB9260, Millipore), PH3 (1:200, 06-570, Millipore), GFAP (1:500, AB5804, Millipore), GLAST (SLC1A3 or EAAT1; 1:200, AB416, Abcam), pSMAD1/5/8 (1:200, #13820, CST), Flag (1:2000, #14793, CST), YAP (1:200, #8418/D24E4, CST), CUX1 (1:200, M222, Santa Cruz), c leaved caspase 3 (1:100, #9661, CST), TBR1 (1:200, AB10554, Millipore) or TBR2 (1:400, AB2283, Millipore), or with monoclonal antibodies against YAP (1:200, WH0010413M1, Sigma), GFAP (1:500, MAB360, Millipore), NeuN (1:500, MAB377, Millipore), MAP2 (1:500, AB5622, Millipore), Tuj1 (1:500, T8660, Sigma) and nestin (1:1000, MAB353, Sigma), or with goat polyclonal antibodies against aldolase C (1:500, SC-12065, Santa Cruz).

    Techniques: Mouse Assay, Double Immunostaining, Cell Culture, BrdU Incorporation Assay, Staining

    Selective expression of YAP in neocortical NSCs and astrocytes. (A) Double immunostaining of YAP (green) and BLBP (red) in the layer III-V neocortex of P1 Yap f/f and Yap nestin -CKO mice. Arrows and arrowheads indicate typical YAP + /BLBP + and YAP + /BLBP − cells, respectively. The boxed region is magnified to the right as single-channel and merged images. (B) Double immunostaining of YAP (red) and aldolase C (green) in the neocortex of P1 Yap f/f and Yap nestin -CKO mice. Arrows indicate the blood vessels. (C,D) Double immunostaining of (C) YAP (green) and nestin (red) and (D) YAP (red) and NeuN (green) in the neocortex of P1 Yap f/f mice. (E,F) Double immunostaining of YAP (green) and GFAP (red) in primary cultured astrocytes with (E) serum-free medium or (F) 10% FBS+DMEM. (G,H) Double immunostaining of (G) YAP (green) and nestin (red) in primary cultured NSCs or (H) YAP (green) and MAP2 (red) in primary cultured neocortical neurons (4 days in vitro ) from WT neocortex. DAPI (blue) was used to stain nuclei. The YAP antibody used in B and D was from CST (#8418/D24E4), otherwise from Sigma (WH0010413M1). Scale bars: 20 μm.

    Journal: Development (Cambridge, England)

    Article Title: YAP stabilizes SMAD1 and promotes BMP2-induced neocortical astrocytic differentiation

    doi: 10.1242/dev.130658

    Figure Lengend Snippet: Selective expression of YAP in neocortical NSCs and astrocytes. (A) Double immunostaining of YAP (green) and BLBP (red) in the layer III-V neocortex of P1 Yap f/f and Yap nestin -CKO mice. Arrows and arrowheads indicate typical YAP + /BLBP + and YAP + /BLBP − cells, respectively. The boxed region is magnified to the right as single-channel and merged images. (B) Double immunostaining of YAP (red) and aldolase C (green) in the neocortex of P1 Yap f/f and Yap nestin -CKO mice. Arrows indicate the blood vessels. (C,D) Double immunostaining of (C) YAP (green) and nestin (red) and (D) YAP (red) and NeuN (green) in the neocortex of P1 Yap f/f mice. (E,F) Double immunostaining of YAP (green) and GFAP (red) in primary cultured astrocytes with (E) serum-free medium or (F) 10% FBS+DMEM. (G,H) Double immunostaining of (G) YAP (green) and nestin (red) in primary cultured NSCs or (H) YAP (green) and MAP2 (red) in primary cultured neocortical neurons (4 days in vitro ) from WT neocortex. DAPI (blue) was used to stain nuclei. The YAP antibody used in B and D was from CST (#8418/D24E4), otherwise from Sigma (WH0010413M1). Scale bars: 20 μm.

    Article Snippet: Primary antibodies were rabbit polyclonal antibodies against nestin (1:200, SAB4200394, Sigma), BLBP (1:300, Ab32423, Abcam), Ki67 (1:200, AB9260, Millipore), PH3 (1:200, 06-570, Millipore), GFAP (1:500, AB5804, Millipore), GLAST (SLC1A3 or EAAT1; 1:200, AB416, Abcam), pSMAD1/5/8 (1:200, #13820, CST), Flag (1:2000, #14793, CST), YAP (1:200, #8418/D24E4, CST), CUX1 (1:200, M222, Santa Cruz), c leaved caspase 3 (1:100, #9661, CST), TBR1 (1:200, AB10554, Millipore) or TBR2 (1:400, AB2283, Millipore), or with monoclonal antibodies against YAP (1:200, WH0010413M1, Sigma), GFAP (1:500, MAB360, Millipore), NeuN (1:500, MAB377, Millipore), MAP2 (1:500, AB5622, Millipore), Tuj1 (1:500, T8660, Sigma) and nestin (1:1000, MAB353, Sigma), or with goat polyclonal antibodies against aldolase C (1:500, SC-12065, Santa Cruz).

    Techniques: Expressing, Double Immunostaining, Mouse Assay, Cell Culture, In Vitro, Staining

    RC2 (red) and BLBP (green) staining of adult uninjured and 10 days pONC mouse coronal samples. Images of the OC and surrounding structures are shown and in (a–aii) adult-uninjured and (c–cii) -injured samples. Scale bar: 500 μm. High-power images of (b–bii) adult-uninjured and (d–dii) -injured samples show the chiasm structure in detail. Blue: DAPI. Scale bar: 100 μm. RC2-positive and BLBP-negative processes were observed extending through the optic chiasm midline (arrows, bii) in uninjured adult mice; while in 10-day pONC adult mice, only BLBP-positive staining was observed (arrows, di).

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Expression of Developmentally Important Axon Guidance Cues in the Adult Optic Chiasm

    doi: 10.1167/iovs.19-26732

    Figure Lengend Snippet: RC2 (red) and BLBP (green) staining of adult uninjured and 10 days pONC mouse coronal samples. Images of the OC and surrounding structures are shown and in (a–aii) adult-uninjured and (c–cii) -injured samples. Scale bar: 500 μm. High-power images of (b–bii) adult-uninjured and (d–dii) -injured samples show the chiasm structure in detail. Blue: DAPI. Scale bar: 100 μm. RC2-positive and BLBP-negative processes were observed extending through the optic chiasm midline (arrows, bii) in uninjured adult mice; while in 10-day pONC adult mice, only BLBP-positive staining was observed (arrows, di).

    Article Snippet: Radial glial markers: RC2 (mouse, 1:5; Developmental Studies Hybridoma Bank), BLBP (rabbit, 1:500, ab32423; Abcam, Cambridge, UK), Slit1 (rabbit, 1:500, PAB11326; AbNova, Taipei, Taiwan) and embryonic development marker Pax2 (rabbit, 1:100, 901001; BioLegend, San Diego, CA, USA) were diluted in blocking solution at 4°C overnight.

    Techniques: Staining, Mouse Assay

    RC2 (red) and BLBP (green) staining of E15.5 mouse coronal samples. (a–aii) Images of the OC and surrounding structures. Scale bar: 500 μm. (b–bii) High-power images show the chiasm structure in detail. (ai) BLBP and (aii) RC2 positive staining was observed immediately superior to the optic chiasm and glial processes (arrows, b) can be observed interacting with the optic chiasm. Blue: DAPI. Scale bar: 100 μm.

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: Expression of Developmentally Important Axon Guidance Cues in the Adult Optic Chiasm

    doi: 10.1167/iovs.19-26732

    Figure Lengend Snippet: RC2 (red) and BLBP (green) staining of E15.5 mouse coronal samples. (a–aii) Images of the OC and surrounding structures. Scale bar: 500 μm. (b–bii) High-power images show the chiasm structure in detail. (ai) BLBP and (aii) RC2 positive staining was observed immediately superior to the optic chiasm and glial processes (arrows, b) can be observed interacting with the optic chiasm. Blue: DAPI. Scale bar: 100 μm.

    Article Snippet: Radial glial markers: RC2 (mouse, 1:5; Developmental Studies Hybridoma Bank), BLBP (rabbit, 1:500, ab32423; Abcam, Cambridge, UK), Slit1 (rabbit, 1:500, PAB11326; AbNova, Taipei, Taiwan) and embryonic development marker Pax2 (rabbit, 1:100, 901001; BioLegend, San Diego, CA, USA) were diluted in blocking solution at 4°C overnight.

    Techniques: Staining

    In cultures of dentate gyrus neural precursor cells virtually all cells express cellular marker for proliferation, Ki67 (A), and for progenitor cells of the nervous system, such as Nestin (A–C), Sox2 (C) or vimentin (D) . Many cells express also the radial glia marker BLBP (B) and Id1 (F) , a dominant-negative bHLH transcriptional regulator expressed in a fraction of adult neural stem cells important for their self-renewal capacity. Only few cells express β3-tubulin (E) indicating neuronally committed cells. No cells were positive for S100β (F) , a marker that labels mature astrocytes. Cells were grown until about 80% confluent (3–5 days) before fixing and staining. Blue stain in (A–F) is the nucleic dye DAPI. Scale bar in (A) : 50 μm, in (A ′ ) and (B) for (A ′ –F) : 20 μm.

    Journal: Frontiers in Neuroscience

    Article Title: A Protocol for Isolation and Enriched Monolayer Cultivation of Neural Precursor Cells from Mouse Dentate Gyrus

    doi: 10.3389/fnins.2011.00089

    Figure Lengend Snippet: In cultures of dentate gyrus neural precursor cells virtually all cells express cellular marker for proliferation, Ki67 (A), and for progenitor cells of the nervous system, such as Nestin (A–C), Sox2 (C) or vimentin (D) . Many cells express also the radial glia marker BLBP (B) and Id1 (F) , a dominant-negative bHLH transcriptional regulator expressed in a fraction of adult neural stem cells important for their self-renewal capacity. Only few cells express β3-tubulin (E) indicating neuronally committed cells. No cells were positive for S100β (F) , a marker that labels mature astrocytes. Cells were grown until about 80% confluent (3–5 days) before fixing and staining. Blue stain in (A–F) is the nucleic dye DAPI. Scale bar in (A) : 50 μm, in (A ′ ) and (B) for (A ′ –F) : 20 μm.

    Article Snippet: 3.2 Reagents Culture medium Neurobasal A (Gibco, cat. no. 10888) or DMEM/F12 (Gibco, cat. no. 31330) for culture dish coating and cell separation procedure B27 Supplement (Gibco, cat. no. 17504) Glutamax-1 Supplement (Gibco, cat. no. 35050) Recombinant human Fibroblast growth factor 2 (FGF2; Peprotech, cat. no. 100-18B-B) Recombinant human Epidermal growth factor (EGF; Peprotech, cat. no. 100-15) Recombinant human Brain Derived Neurotrophic Factor (BDNF; Peprotech, cat. no. 450-02) Bovine serum albumin (BSA; Sigma, cat. no. A2153) d -Glucose (Sigma, cat. no. G7021) Retinoic acid (Sigma, cat. no. R2625) Cell detachment Accutase (PAA, cat. no. L11-007) Ovomucoid trypsin inhibitor (Worthington, cat. no. 3085) Trypsin EDTA 10× (Gibco, cat. no. 15400054) Cell extraction Papain (Worthington, cat. no. 3126) Dispase (Roche, cat. no. 10-165-859-001) DNase (Worthington, cat. no. 2139) Percoll (GE Healthcare, cat. no. 17-0891-01) PBS 10× (Gibco, cat. no. 70011) Coating Laminin (Roche, cat. no. 11-243-217-001) Poly-d -Lysine (PDL; Sigma, cat. no. P1024) Ethanol 70% vol/vol PBS (Gibco, cat. no. 10010-023) H2 SO4 (EMD, cat. no. SX1244-6) ddH2 O Cell banking Dimethyl sulfoxide (DMSO; Sigma Aldrich, cat. no. D8418) Immunocytochemistry Paraformaldehyde (PFA; Electron Microscopy Sciences, cat. no. 19210) Triton-X100 (Sigma, cat. no. T8787) Normal donkey serum (Jackson Immunoresearch, cat. no. 017-000-121) Hoechst 33258 (Gibco, cat. no. H3569) Mouse anti-Map2ab antibody (Sigma, cat. no. M1406; 1:500) Rabbit anti-GFAP antibody (Dako, cat. no. Z0334; 1:500) Mouse anti-O4 antibody (R & D Systems, cat. no. MAB1326; 1:100) Mouse anti-Nestin antibody (BD Pharmingen, cat. no. 611658; 1:200) Rabbit anti-Sox2 antibody (Chemicon, cat. no. AB5603; 1:500) Rabbit anti-Ki67 antibody (Novocastra, cat. no. NCL-Ki67p; 1:400) Rabbit anti-BLBP antibody (Abcam, cat. no. ab32423; 1:400) Goat anti-Vimentin antibody (Sigma, cat. no. V4630; 1:500) Rabbit anti-Id1 antibody (Biocheck, cat. no. BCH-1/37-2; 1:100) Mouse anti-β3-tubulin antibody (Promega, cat. no. G712A; 1:1000) Rabbit anti-S100b antibody (Swant, cat. no. 37A; 1:1000)

    Techniques: Marker, Dominant Negative Mutation, Staining

    Characterization of the brain endocannabinoid system in FABP5/7 KO mice. a RT-PCR analysis of FABP expression in brains of WT and FABP5/7 KO mice. Note that of the ten FABP isoform profiled, selective deletion of FABP5 and FABP7 was observed in the brains of FABP5/7 KO mice. b Western blot confirms the absence of FABP5 and FABP7 in the brains of FABP5/7 KO mice. c Brain PEA, OEA, AEA, and 2-AG levels in WT and FABP5/7 KO mice. *p

    Journal: Molecular Pain

    Article Title: Fatty acid binding protein deletion suppresses inflammatory pain through endocannabinoid/N-acylethanolamine-dependent mechanisms

    doi: 10.1186/s12990-015-0056-8

    Figure Lengend Snippet: Characterization of the brain endocannabinoid system in FABP5/7 KO mice. a RT-PCR analysis of FABP expression in brains of WT and FABP5/7 KO mice. Note that of the ten FABP isoform profiled, selective deletion of FABP5 and FABP7 was observed in the brains of FABP5/7 KO mice. b Western blot confirms the absence of FABP5 and FABP7 in the brains of FABP5/7 KO mice. c Brain PEA, OEA, AEA, and 2-AG levels in WT and FABP5/7 KO mice. *p

    Article Snippet: Blots were probed with the following antibodies: COX2 (1:1000, Abcam #Ab15191), FAAH (1:1000, Abcam #Ab54615), MAGL (1:400, Cayman Chemical #10212), GAPDH (1:5000, Abcam #Ab8245), CB1 (1:1000, Abcam #Ab172970), NAPE-PLD (1:400, Abcam #Ab95397), FABP5 (1:1000, BioVendor R & D #RD181060100), or FABP7 (1:200, Abcam #Ab32423).

    Techniques: Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

    CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker BLBP (A’, red). Merged images (A ″ ). Immuno-staining for GFAP (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).

    Journal: Hippocampus

    Article Title: Identification of a Sustained Neurogenic Zone at the Dorsal Surface of the Adult Mouse Hippocampus and Its Regulation by the Chemokine SDF-1

    doi: 10.1002/hipo.22428

    Figure Lengend Snippet: CXCR4-EGFP cells along the aCMS pathway have migratory appearance and express stem/neural progenitor markers. A–H: Representative confocal images showing CXCR4-EGFP cells (green) with morphologies resembling migratory progenitors, in which a large proportion have un or bipolar morphologies, indicating their potential to migrate. A-A ″ : Example of CXCR4-EGFP cells with one or two long processes (green, A) stained for the radial glia marker BLBP (A’, red). Merged images (A ″ ). Immuno-staining for GFAP (B, B’, B ″ ) and nestin (C, C’, C ″ ) showed the coexpression of GFAP (red) and nestin (red) in GFP cells (green), in the fimbria (B, C), and along the meninges (B’, C’), some with an apical radial glia process (arrows) with multiple endings at the meningeal-DG junction (B ″ , C ″ ). Similarly, cells in the fimbria (D), along the meninges (D’) and at the meningeal-DG junction also stained for the neuronal stem marker SOX-2 (red), including cells at the meningeal-DG junction (D ″ ). Immunostaining for the neuronal progenitor marker Tbr-2, which showed it expression around the cell bodies and in the cell processes (E’), revealed that some of the cells in the SHZ and virtually all migratory cells within the fimbria (arrows, E, E ″ ) stained for Tbr-2 (arrows, E’, E ″ ).

    Article Snippet: Briefly, sections were blocked in phosphate buffer containing 0.1% Triton X-100 and incubated overnight at 4 °C with the following primary antibodies: CD45 (1/300, rat, Millipore, CA) and Iba-1 (1/300, rabbit, Wako Chemicals USA, VA) for microglia; CD45 and F4/80 (1/300, rabbit, Santa Cruz Biotechnology) for macrophages; Nestin (1/150, rat, BD Pharmingen, CA) for early neural progenitors, SOX-2 (1/200, rabbit, Millipore, CA) for neuronal stem cells, GFAP (1/300, mouse, Sigma-Aldrich, MO) and BLBP (1/100, rabbit, Millipore) for radial glia, DCX [1:700; Guina pig, Millipore, CA) for migratory neuroblasts, NeuN (1/300, mouse, Milli-pore, MA), Prox-1 (1/500, rabbit, Millipore, CA)] for DG granular neurons, calretinin and calbindin (1/250, rabbit, Millipore, MA) for mature DG neurons, laminin (1/100, rabbit, Millipore, CA), vWF (1/100, rabbit, Santa Cruz Biotechnology, CA) for blood vessels.

    Techniques: Staining, Marker, Immunostaining, Expressing

    RT-PCR analysis Expression level of OTX2, cMYC, OCT3/4, Nestin, Vimentin, Sox2 and BLBP in the RG and TCL self-renewing cells.

    Journal: Oncotarget

    Article Title: Characterization of brain tumor initiating cells isolated from an animal model of CNS primitive neuroectodermal tumors

    doi: 10.18632/oncotarget.24460

    Figure Lengend Snippet: RT-PCR analysis Expression level of OTX2, cMYC, OCT3/4, Nestin, Vimentin, Sox2 and BLBP in the RG and TCL self-renewing cells.

    Article Snippet: The immunohistochemical panel comprised the following antibodies: Anti-Ki-67 (RM-9106, Rabbit monoclonal, 1:200, Thermo scientific), OCT3/4 (H-134) (sc-9081, Rabbit polyclonal, 1:50, Santa Cruz), Anti-Nestin (ab105389, Rabbit monoclonal, 1:30, abcam), Anti-Sox2 (ab97959, Rabbit polyclonal, 1:450, abcam), Anti-Vimentin (NBP1-97671, Mouse monoclonal, 1:500, Novus Biologicals), c-MYC (ab32072, Rabbit monoclonal[Y69], 1:500, abcam), Anti-c-MYC-(Phospho S62) (ab185656, Rabbit monoclonal, 1:500, abcam), p53 (FL-393) (sc-6243, Rabbit polyclonal, 1:200, Santa Cruz), Anti-BLBP (ABN14, Rabbit polyclonal, 1:400, Millipore), Anti-OTX2 (AB9566, Rabbit polyclonal, 1:750, Millipore), Anti-Trim22 (ab140966, Mouse monoclonal, 1:100, abcam), Caveolin (N-20) (sc-894, Rabbit polyclonal, 1:50, Santa Cruz), Cathepsin C/DPPI (AF1034, Goat polyclonal, 1:50, R & D), ENO1 (LS-B10960, Rabbit polyclonal, 1:500, LSBio), Anti-MALT1 (ab93661, Rabbit polyclonal, 1:200, abcam), EPHA3 (LS-C312723, Rabbit polyclonal, 1:200, LSBio), Anti-PRAME (ab135600, Rabbit polyclonal, 1:150, abcam), Anti-MDM2 (LS-C199239, Rabbit polyclonal, 1:100, LS Bio).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

    Analysis of RG, TCL and the TCL derived tumors (A1, A2) LCAS-R 2nd nsphrs/LCAS-RTL(138) 2nd nsphr, (A3) Tubb3 40x LCAS-RTL(138) 2nd nsphr, (A4) GFAP 40x LCAS-RTL(138) 2nd nsphr; (B1, B2) HE 40x LCAS-R/LCAS-RTL(138) - images show undifferentiated small round blue cells with scarce cytoplasm, (B3) HE 40x (LCAS-RTL(138) SVZ 15 weeks post-injection) - extensive proliferation of undifferentiated embryonal tumor cells infiltrating the adjacent brain parenchyma, (B4) Ki67 40x (LCAS-RTL(138) SVZ 15 weeks post-injection) - high proliferative activity of tumor cells is reflected by over 75% of cells expressing Ki-67, (C1) OCT3/4 40x (LC26-RTL(170) SVZ 12 weeks post-injection) - extensive proliferation of tumor cells invading adjacent brain parenchyma show high expression of OCT3/4 (over 80% of cells), (C2) Sox2 40x (LC26-RTL(170) SVZ 12 weeks post-injection) - extensively expressed in the tumor cells, (C3) PRAME 40x (LCAS-R 12 weeks post-injection) - extensively expressed in the tumor cells, (C4) Nestin 40x (LCAS-RTL(138) 15 weeks post-injection) - extensively expressed in the tumor cells, (D1) Vimentin 40x (LCAS-RTL(138) 15 weeks post-injection) - extensively expressed in the tumor cells within the main tumor mass and in the invading areas, (D2) BLBP 40x (LC26-RTL(170) 16 weeks post-injection) - extensively expressed in the tumor cells, (D3, D4) OTX2 20x, 40x (LC26-RTL(170) 16 weeks post-injection) - extensive expression (over 80% of cells) in tumor cells. TU-tumor; P-parenchyma; PI- perivascular invasion.

    Journal: Oncotarget

    Article Title: Characterization of brain tumor initiating cells isolated from an animal model of CNS primitive neuroectodermal tumors

    doi: 10.18632/oncotarget.24460

    Figure Lengend Snippet: Analysis of RG, TCL and the TCL derived tumors (A1, A2) LCAS-R 2nd nsphrs/LCAS-RTL(138) 2nd nsphr, (A3) Tubb3 40x LCAS-RTL(138) 2nd nsphr, (A4) GFAP 40x LCAS-RTL(138) 2nd nsphr; (B1, B2) HE 40x LCAS-R/LCAS-RTL(138) - images show undifferentiated small round blue cells with scarce cytoplasm, (B3) HE 40x (LCAS-RTL(138) SVZ 15 weeks post-injection) - extensive proliferation of undifferentiated embryonal tumor cells infiltrating the adjacent brain parenchyma, (B4) Ki67 40x (LCAS-RTL(138) SVZ 15 weeks post-injection) - high proliferative activity of tumor cells is reflected by over 75% of cells expressing Ki-67, (C1) OCT3/4 40x (LC26-RTL(170) SVZ 12 weeks post-injection) - extensive proliferation of tumor cells invading adjacent brain parenchyma show high expression of OCT3/4 (over 80% of cells), (C2) Sox2 40x (LC26-RTL(170) SVZ 12 weeks post-injection) - extensively expressed in the tumor cells, (C3) PRAME 40x (LCAS-R 12 weeks post-injection) - extensively expressed in the tumor cells, (C4) Nestin 40x (LCAS-RTL(138) 15 weeks post-injection) - extensively expressed in the tumor cells, (D1) Vimentin 40x (LCAS-RTL(138) 15 weeks post-injection) - extensively expressed in the tumor cells within the main tumor mass and in the invading areas, (D2) BLBP 40x (LC26-RTL(170) 16 weeks post-injection) - extensively expressed in the tumor cells, (D3, D4) OTX2 20x, 40x (LC26-RTL(170) 16 weeks post-injection) - extensive expression (over 80% of cells) in tumor cells. TU-tumor; P-parenchyma; PI- perivascular invasion.

    Article Snippet: The immunohistochemical panel comprised the following antibodies: Anti-Ki-67 (RM-9106, Rabbit monoclonal, 1:200, Thermo scientific), OCT3/4 (H-134) (sc-9081, Rabbit polyclonal, 1:50, Santa Cruz), Anti-Nestin (ab105389, Rabbit monoclonal, 1:30, abcam), Anti-Sox2 (ab97959, Rabbit polyclonal, 1:450, abcam), Anti-Vimentin (NBP1-97671, Mouse monoclonal, 1:500, Novus Biologicals), c-MYC (ab32072, Rabbit monoclonal[Y69], 1:500, abcam), Anti-c-MYC-(Phospho S62) (ab185656, Rabbit monoclonal, 1:500, abcam), p53 (FL-393) (sc-6243, Rabbit polyclonal, 1:200, Santa Cruz), Anti-BLBP (ABN14, Rabbit polyclonal, 1:400, Millipore), Anti-OTX2 (AB9566, Rabbit polyclonal, 1:750, Millipore), Anti-Trim22 (ab140966, Mouse monoclonal, 1:100, abcam), Caveolin (N-20) (sc-894, Rabbit polyclonal, 1:50, Santa Cruz), Cathepsin C/DPPI (AF1034, Goat polyclonal, 1:50, R & D), ENO1 (LS-B10960, Rabbit polyclonal, 1:500, LSBio), Anti-MALT1 (ab93661, Rabbit polyclonal, 1:200, abcam), EPHA3 (LS-C312723, Rabbit polyclonal, 1:200, LSBio), Anti-PRAME (ab135600, Rabbit polyclonal, 1:150, abcam), Anti-MDM2 (LS-C199239, Rabbit polyclonal, 1:100, LS Bio).

    Techniques: Derivative Assay, Injection, Activity Assay, Expressing

    Comparative analysis of reprogramming potentials of distinct factor combinations. (A) The schematic illustration depicting the strategy for comparing the reprogramming efficiency of distinct factor combinations. (B) Time-course immunofluorescence analysis for comparing reprogramming potentials of distinct factor combinations. Scale bars, 100 μ m. (C) Morphology of hiNSC clusters at 2 weeks after transduction. Scale bars, 100 μ m. (D) The number of BLBP + / MSI1 + colonies were counted in a time-course manner. Data are presented as mean±SD from six independent experiments. *p

    Journal: International Journal of Stem Cells

    Article Title: Robust and Reproducible Generation of Induced Neural Stem Cells from Human Somatic Cells by Defined Factors

    doi: 10.15283/ijsc19097

    Figure Lengend Snippet: Comparative analysis of reprogramming potentials of distinct factor combinations. (A) The schematic illustration depicting the strategy for comparing the reprogramming efficiency of distinct factor combinations. (B) Time-course immunofluorescence analysis for comparing reprogramming potentials of distinct factor combinations. Scale bars, 100 μ m. (C) Morphology of hiNSC clusters at 2 weeks after transduction. Scale bars, 100 μ m. (D) The number of BLBP + / MSI1 + colonies were counted in a time-course manner. Data are presented as mean±SD from six independent experiments. *p

    Article Snippet: The following primary antibodies were used: goat anti-SOX2 (Santa Cruz, 1:200), goat anti BRN2 (Santa Cruz, 1:200), rabbit anti-BLBP (Santa Cruz, 1:200), rat anti-MSI1 (MBL, 1:200), rabbit anti-MSI2 (Abcam, 1:200), mouse anti-TUJ1 (Covance, 1:500), rabbit anti-GFAP (Dako, 1:500), mouse anti-MBP (Abcam, 1:500), rabbit anti-GABA (Sigma, 1:200), rabbit anti-GLU (Sigma, 1:200), goat-anti ChAT (Merck, 1:200), and rabbit anti-TH (Merck, 1:200),and rabbit anti-MBP (Abcam, 1:200).

    Techniques: Immunofluorescence, Transduction

    Characterization of SKM LT hiNSCs. (A) Immunofluorescence images of hESC-derived NSCs and SKM LT hiNSCs using antibodies against BRN2 , BLBP , MSI1 , and MSI2 . Scale bars, 100 μ m. (B) A heat map representing expression profile of genes with more than two-fold expression level difference between hFFs and hESC-derived NSCs. The color represents z-score for gene expression level in log 2 scale. Clusters with red and blue bar on the left represent genes with lower and higher expression in hFFs compared to hESC-derived NSCs, respectively. (C, D) Differentiation potential of SKM LT hiNSCs into astrocytes (C) and neurons (D) as determined by immunocytochemistry with antibodies against GFAP and TUJ1 , respectively, Scale bars, 100 μ m. (E, F) The efficiency of differentiation into astrocytes (E) and neurons (F) from hESC-derived NSCs and SKM LT hiNSCs was quantified and compared via immunostaining with GFAP and TUJ1 , respectively. Data are presented as mean±SD from eight independent experiments. N.S.: not significant.

    Journal: International Journal of Stem Cells

    Article Title: Robust and Reproducible Generation of Induced Neural Stem Cells from Human Somatic Cells by Defined Factors

    doi: 10.15283/ijsc19097

    Figure Lengend Snippet: Characterization of SKM LT hiNSCs. (A) Immunofluorescence images of hESC-derived NSCs and SKM LT hiNSCs using antibodies against BRN2 , BLBP , MSI1 , and MSI2 . Scale bars, 100 μ m. (B) A heat map representing expression profile of genes with more than two-fold expression level difference between hFFs and hESC-derived NSCs. The color represents z-score for gene expression level in log 2 scale. Clusters with red and blue bar on the left represent genes with lower and higher expression in hFFs compared to hESC-derived NSCs, respectively. (C, D) Differentiation potential of SKM LT hiNSCs into astrocytes (C) and neurons (D) as determined by immunocytochemistry with antibodies against GFAP and TUJ1 , respectively, Scale bars, 100 μ m. (E, F) The efficiency of differentiation into astrocytes (E) and neurons (F) from hESC-derived NSCs and SKM LT hiNSCs was quantified and compared via immunostaining with GFAP and TUJ1 , respectively. Data are presented as mean±SD from eight independent experiments. N.S.: not significant.

    Article Snippet: The following primary antibodies were used: goat anti-SOX2 (Santa Cruz, 1:200), goat anti BRN2 (Santa Cruz, 1:200), rabbit anti-BLBP (Santa Cruz, 1:200), rat anti-MSI1 (MBL, 1:200), rabbit anti-MSI2 (Abcam, 1:200), mouse anti-TUJ1 (Covance, 1:500), rabbit anti-GFAP (Dako, 1:500), mouse anti-MBP (Abcam, 1:500), rabbit anti-GABA (Sigma, 1:200), rabbit anti-GLU (Sigma, 1:200), goat-anti ChAT (Merck, 1:200), and rabbit anti-TH (Merck, 1:200),and rabbit anti-MBP (Abcam, 1:200).

    Techniques: Immunofluorescence, Derivative Assay, Expressing, Immunocytochemistry, Immunostaining

    Genome wide correlation analysis between Ca 2+ drug sensitivity and gene expression. (A) Nine novel GIC lines were subjected to Thapsigargin dose response analysis (0.1–100 µM), showing different response to moderate drug doses. (B, C) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and (B) NES and (C) FABP7/BLBP mRNA expression. U3047-MG was considered an outlier in the NES graph (marked in red) and excluded form the analysis. (D) Western blot analysis showing BLBP (FABP7) protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines, with β-actin as loading control. (E) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and GRIA1 mRNA expression. (F) Western blot analysis showing GRIA1 protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines. β-actin was used as loading control.

    Journal: PLoS ONE

    Article Title: Selective Calcium Sensitivity in Immature Glioma Cancer Stem Cells

    doi: 10.1371/journal.pone.0115698

    Figure Lengend Snippet: Genome wide correlation analysis between Ca 2+ drug sensitivity and gene expression. (A) Nine novel GIC lines were subjected to Thapsigargin dose response analysis (0.1–100 µM), showing different response to moderate drug doses. (B, C) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and (B) NES and (C) FABP7/BLBP mRNA expression. U3047-MG was considered an outlier in the NES graph (marked in red) and excluded form the analysis. (D) Western blot analysis showing BLBP (FABP7) protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines, with β-actin as loading control. (E) Plot of correlation between cell viability after Ca 2+ drug exposure (Thapsigargin, 1 uM) and GRIA1 mRNA expression. (F) Western blot analysis showing GRIA1 protein expression in selected Thapsigargin sensitive (U3017-MG and U3084-MG) and less sensitive (U3013-MG and U3035-MG) cell lines. β-actin was used as loading control.

    Article Snippet: Membranes were blocked in 5% milk for 1 hour and incubated overnight at 4°C with the following primary antibodies: rabbit anti-GRIA1 antibody (Abcam), rabbit anti-S100 alpha 6 antibody (Abcam), mouse anti-BLBP antibody (Millipore) and mouse anti-beta actin antibody (Abcam).

    Techniques: Genome Wide, Expressing, Western Blot