rabbit anti mouse igg  (Valiant)

 
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    Name:
    Anti mouse IgG whole molecule rabbit antiserum
    Description:
    Product is the lyophilized powder of rabbit antiserum to mouse IgG whole molecule and buffer salts
    Catalog Number:
    0855436
    Price:
    95.25
    Category:
    Life Sciences Antibodies Antisera
    Applications:
    Immunoassays
    Size:
    2 mL
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    Structured Review

    Valiant rabbit anti mouse igg
    Anti mouse IgG whole molecule rabbit antiserum
    Product is the lyophilized powder of rabbit antiserum to mouse IgG whole molecule and buffer salts
    https://www.bioz.com/result/rabbit anti mouse igg/product/Valiant
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mouse igg - by Bioz Stars, 2021-05
    93/100 stars

    Images

    1) Product Images from "Inactivation of the RB family prevents thymus involution and promotes thymic function by direct control of Foxn1 expression"

    Article Title: Inactivation of the RB family prevents thymus involution and promotes thymic function by direct control of Foxn1 expression

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20121716

    The RB family controls the expression of Foxn1 . (A) Expression of Foxn1 in CD45 – MHCII + TECS from control (Ctrl) and mutant (Mut) mice relative to Eva1 expression ( n = 4; P = 0.03). (B) Immunoblot analysis of Foxn1 levels in control and mutant TECs from 12 pooled control and 5 pooled mutant thymi. α-Tubulin is included as a loading control ( n = 2). (C) Immunofluorescence staining of Foxn1 and CD205 individually and merged (right) in control (top) and mutant (bottom) thymi (one of two shown). Bars, 100 µm. (D) Schematic representation of the Foxn1 promoter. Region 1a is active in the skin and thymus, whereas region 1b is only active in the skin. (E) Sequence alignment on the UCSC genome browser ( Fujita et al., 2011 ) of the promoter region 1a encompassing two E2F sites shows conservation across multiple mammalian species ( Kent, 2002 ). Gray highlighting indicates conserved sequences, and black highlighting indicates putative E2F binding sites. (F) Transfection into 210R TECs of an expression plasmid coding for E2F3 together with a luciferase reporter plasmid for either the wild-type region 1a of the Foxn1 promoter or a region 1a where site 3 has been mutated ( n = 3; P = 0.03). (G) Transfection into 210R cells of E2F3 and RB expression and the Foxn1 luciferase reporter plasmid ( n = 3; P = 0.02). (H) ChIP analysis of E2F3 and E2F4 to sites 1 and 2–3 in TEC100 cells. To adjust for inter-experiment variability of scale of percent input, experiments were normalized to nonspecific antibodies (IgG and anti-p16 INK4a ). This ratio is represented here. In addition, binding to a nonspecific region of the fifth chromosome was included as a negative control ( n = 3; for site 1 P = 0.02, for site 2–3 P = 0.04). (I) Fold change in percent input as assessed by ChIP analysis of E2F3 and E2F4 binding to the Foxn1 promoter in TEC100 cells after transient overexpression of RB ( n = 3 compared with untransfected controls; p-values: E2F3 Foxn1 site 1 = 0.06 and site 2–3 = 0.04; E2F4 Foxn1 site 1 = 0.01 and site 2–3 = 0.02). Asterisks indicate P
    Figure Legend Snippet: The RB family controls the expression of Foxn1 . (A) Expression of Foxn1 in CD45 – MHCII + TECS from control (Ctrl) and mutant (Mut) mice relative to Eva1 expression ( n = 4; P = 0.03). (B) Immunoblot analysis of Foxn1 levels in control and mutant TECs from 12 pooled control and 5 pooled mutant thymi. α-Tubulin is included as a loading control ( n = 2). (C) Immunofluorescence staining of Foxn1 and CD205 individually and merged (right) in control (top) and mutant (bottom) thymi (one of two shown). Bars, 100 µm. (D) Schematic representation of the Foxn1 promoter. Region 1a is active in the skin and thymus, whereas region 1b is only active in the skin. (E) Sequence alignment on the UCSC genome browser ( Fujita et al., 2011 ) of the promoter region 1a encompassing two E2F sites shows conservation across multiple mammalian species ( Kent, 2002 ). Gray highlighting indicates conserved sequences, and black highlighting indicates putative E2F binding sites. (F) Transfection into 210R TECs of an expression plasmid coding for E2F3 together with a luciferase reporter plasmid for either the wild-type region 1a of the Foxn1 promoter or a region 1a where site 3 has been mutated ( n = 3; P = 0.03). (G) Transfection into 210R cells of E2F3 and RB expression and the Foxn1 luciferase reporter plasmid ( n = 3; P = 0.02). (H) ChIP analysis of E2F3 and E2F4 to sites 1 and 2–3 in TEC100 cells. To adjust for inter-experiment variability of scale of percent input, experiments were normalized to nonspecific antibodies (IgG and anti-p16 INK4a ). This ratio is represented here. In addition, binding to a nonspecific region of the fifth chromosome was included as a negative control ( n = 3; for site 1 P = 0.02, for site 2–3 P = 0.04). (I) Fold change in percent input as assessed by ChIP analysis of E2F3 and E2F4 binding to the Foxn1 promoter in TEC100 cells after transient overexpression of RB ( n = 3 compared with untransfected controls; p-values: E2F3 Foxn1 site 1 = 0.06 and site 2–3 = 0.04; E2F4 Foxn1 site 1 = 0.01 and site 2–3 = 0.02). Asterisks indicate P

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, Immunofluorescence, Staining, Sequencing, Binding Assay, Transfection, Plasmid Preparation, Luciferase, Chromatin Immunoprecipitation, Negative Control, Over Expression

    Related Articles

    Purification:

    Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity
    Article Snippet: .. Reagents Various reagents were purchased from following companies: mouse monoclonal antibodies anti-CD3 (145-2C11), anti-CD4 (GK1.5), anti-CD8 (53–6.7), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CCR7 (4B12), anti-CD11c (N418), anti-CD40 (3/23), anti-CD80 (16-10A1), anti-CD86 (GL-1), anti-I-Ab (AF6-120.1), anti-IL-2 (JES6-5H4), anti-IFN-γ (XMG1.2), anti-TNF-α (MP6-XT22) from Biolegend (San Diego, CA), purified anti-mouse IL-12 p70 (C18.2) and biotin anti-mouse IL-12/IL-23 p40 (C17.8) from Biolegend, purified anti-mouse IFN-γ (R4-6A2) and biotin anti-mouse IFN-γ (XMG1.2) from Biolegend, ELISA set of murine IL-1β from BD Bioscience (San Jose, CA), 10x RBC lysis buffer and fixation/permeabilization buffers from eBioscience (San Diego, CA), Alexa Fluor 488® protein labeling kit from Invitrogen (Grand Island, NY), HRP-conjugated goat anti-mouse IgG, IgG1 and rabbit anti-mouse IgG2a, 2b from MP Biomedicals (Solon, OH, USA), and Polymyxin B from Sigma-Aldrich (St. Louis, MO). .. CD4+ T cell epitope (FYQEAAPAQSDVAL) targeting NoV GII.4 strain VA387 were predicted based on T cell epitope prediction tools from Immune Epitope Database (IEDB) Analysis Resource (website: http://tools.immuneepitope.org/main/ ) and synthesized by GenScript (Piscataway, NJ).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity
    Article Snippet: .. Reagents Various reagents were purchased from following companies: mouse monoclonal antibodies anti-CD3 (145-2C11), anti-CD4 (GK1.5), anti-CD8 (53–6.7), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CCR7 (4B12), anti-CD11c (N418), anti-CD40 (3/23), anti-CD80 (16-10A1), anti-CD86 (GL-1), anti-I-Ab (AF6-120.1), anti-IL-2 (JES6-5H4), anti-IFN-γ (XMG1.2), anti-TNF-α (MP6-XT22) from Biolegend (San Diego, CA), purified anti-mouse IL-12 p70 (C18.2) and biotin anti-mouse IL-12/IL-23 p40 (C17.8) from Biolegend, purified anti-mouse IFN-γ (R4-6A2) and biotin anti-mouse IFN-γ (XMG1.2) from Biolegend, ELISA set of murine IL-1β from BD Bioscience (San Jose, CA), 10x RBC lysis buffer and fixation/permeabilization buffers from eBioscience (San Diego, CA), Alexa Fluor 488® protein labeling kit from Invitrogen (Grand Island, NY), HRP-conjugated goat anti-mouse IgG, IgG1 and rabbit anti-mouse IgG2a, 2b from MP Biomedicals (Solon, OH, USA), and Polymyxin B from Sigma-Aldrich (St. Louis, MO). .. CD4+ T cell epitope (FYQEAAPAQSDVAL) targeting NoV GII.4 strain VA387 were predicted based on T cell epitope prediction tools from Immune Epitope Database (IEDB) Analysis Resource (website: http://tools.immuneepitope.org/main/ ) and synthesized by GenScript (Piscataway, NJ).

    Lysis:

    Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity
    Article Snippet: .. Reagents Various reagents were purchased from following companies: mouse monoclonal antibodies anti-CD3 (145-2C11), anti-CD4 (GK1.5), anti-CD8 (53–6.7), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CCR7 (4B12), anti-CD11c (N418), anti-CD40 (3/23), anti-CD80 (16-10A1), anti-CD86 (GL-1), anti-I-Ab (AF6-120.1), anti-IL-2 (JES6-5H4), anti-IFN-γ (XMG1.2), anti-TNF-α (MP6-XT22) from Biolegend (San Diego, CA), purified anti-mouse IL-12 p70 (C18.2) and biotin anti-mouse IL-12/IL-23 p40 (C17.8) from Biolegend, purified anti-mouse IFN-γ (R4-6A2) and biotin anti-mouse IFN-γ (XMG1.2) from Biolegend, ELISA set of murine IL-1β from BD Bioscience (San Jose, CA), 10x RBC lysis buffer and fixation/permeabilization buffers from eBioscience (San Diego, CA), Alexa Fluor 488® protein labeling kit from Invitrogen (Grand Island, NY), HRP-conjugated goat anti-mouse IgG, IgG1 and rabbit anti-mouse IgG2a, 2b from MP Biomedicals (Solon, OH, USA), and Polymyxin B from Sigma-Aldrich (St. Louis, MO). .. CD4+ T cell epitope (FYQEAAPAQSDVAL) targeting NoV GII.4 strain VA387 were predicted based on T cell epitope prediction tools from Immune Epitope Database (IEDB) Analysis Resource (website: http://tools.immuneepitope.org/main/ ) and synthesized by GenScript (Piscataway, NJ).

    Labeling:

    Article Title: Norovirus P Particle Efficiently Elicits Innate, Humoral and Cellular Immunity
    Article Snippet: .. Reagents Various reagents were purchased from following companies: mouse monoclonal antibodies anti-CD3 (145-2C11), anti-CD4 (GK1.5), anti-CD8 (53–6.7), anti-CD44 (IM7), anti-CD62L (MEL-14), anti-CCR7 (4B12), anti-CD11c (N418), anti-CD40 (3/23), anti-CD80 (16-10A1), anti-CD86 (GL-1), anti-I-Ab (AF6-120.1), anti-IL-2 (JES6-5H4), anti-IFN-γ (XMG1.2), anti-TNF-α (MP6-XT22) from Biolegend (San Diego, CA), purified anti-mouse IL-12 p70 (C18.2) and biotin anti-mouse IL-12/IL-23 p40 (C17.8) from Biolegend, purified anti-mouse IFN-γ (R4-6A2) and biotin anti-mouse IFN-γ (XMG1.2) from Biolegend, ELISA set of murine IL-1β from BD Bioscience (San Jose, CA), 10x RBC lysis buffer and fixation/permeabilization buffers from eBioscience (San Diego, CA), Alexa Fluor 488® protein labeling kit from Invitrogen (Grand Island, NY), HRP-conjugated goat anti-mouse IgG, IgG1 and rabbit anti-mouse IgG2a, 2b from MP Biomedicals (Solon, OH, USA), and Polymyxin B from Sigma-Aldrich (St. Louis, MO). .. CD4+ T cell epitope (FYQEAAPAQSDVAL) targeting NoV GII.4 strain VA387 were predicted based on T cell epitope prediction tools from Immune Epitope Database (IEDB) Analysis Resource (website: http://tools.immuneepitope.org/main/ ) and synthesized by GenScript (Piscataway, NJ).

    Incubation:

    Article Title: A Conserved Organization of Transcription during Embryonic Stem Cell Differentiation and in Cells with High C Value
    Article Snippet: The estimated average volume of 10 fluorescent microspheres (6 and 15 μm in diameter; Molecular Probes) lay within 1% of the real volume. .. Br-RNA in cryosections on grids was incubated with the primary anti-BrdU, washed (all as described above) and incubated with either 1) goat anti-mouse IgG conjugated with 5-nm gold particles (1 μg/ml; British Biocell International, Cardiff, United Kingdom) for 3–4 h or 2) rabbit anti-mouse IgG (13 μg/ml; MP Biomedicals, Irvine, CA) for 1 h followed by goat anti-rabbit IgG conjugated with 5-nm particles (1 μg/ml; British Biocell International) for 3–4 h. Because double and triple sandwiches gave similar site densities and diameters, results were pooled. .. After the final antibody, cryosections were washed (4 times over 40 min) in PBS+, rewashed (16 h; 4°C) in PBS and then in water (5 drops; 1 h), and stained (10 min; 4°C) with 0.3% uranyl acetate in 2% methylcellulose.

    Article Title: Diverse molecular mechanisms contribute to differential expression of human duplicated genes
    Article Snippet: ChIP enrichment was performed by incubation for 16 h at 4°C with the following antibodies: 2 μg H3K27ac antibody (Active Motif #39133), 4 μg H3K4me1 antibody (Millipore 07-436), 2 μg H3K4me3 antibody (Active Motif #39915), or 2 μg RNA Polymerase II (PolII) antibody clone 8WG16 (Covance MMS-126R). .. RNA PolII samples were incubated for an additional hour with 2 μg Rabbit Anti-Mouse IgG (MP Biomedical #55436). .. Immune complexes were bound to 20 μl magnetic protein A/G beads (ThermoFisher) for 2 hours at 4°C.

    Article Title: Inactivation of the RB family prevents thymus involution and promotes thymic function by direct control of Foxn1 expression
    Article Snippet: Chromatin was sonicated using a fisher probe sonicator 30% output power for eight cycles of 30 s. The chromatin was precleared before being diluted and bound by 4 µg of the primary antibody overnight at 4°C. .. Each ChIP was then incubated with 8 µg rabbit anti–mouse IgG (MP Biomedicals) or anti–mouse p16 antibody (N-20; Santa Cruz Biotechnology, Inc.) as a secondary for 1 h. Nucleoprotein complexes were pulled down using Pansorbin cells (EMD Millipore). .. Complexes were digested with Proteinase K and RNase A and purified by a QIAquick PCR Purification kit (QIAGEN).

    Chromatin Immunoprecipitation:

    Article Title: Inactivation of the RB family prevents thymus involution and promotes thymic function by direct control of Foxn1 expression
    Article Snippet: Chromatin was sonicated using a fisher probe sonicator 30% output power for eight cycles of 30 s. The chromatin was precleared before being diluted and bound by 4 µg of the primary antibody overnight at 4°C. .. Each ChIP was then incubated with 8 µg rabbit anti–mouse IgG (MP Biomedicals) or anti–mouse p16 antibody (N-20; Santa Cruz Biotechnology, Inc.) as a secondary for 1 h. Nucleoprotein complexes were pulled down using Pansorbin cells (EMD Millipore). .. Complexes were digested with Proteinase K and RNase A and purified by a QIAquick PCR Purification kit (QIAGEN).

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  • 92
    Valiant rabbit anti β galactosidase
    Segregated distribution of D1R- and D2R/A2aR-expressing MSNs in the caudal part of the striatum . (A) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd2-EGFP mice ( n = 10). Scale bar, 200 μm. High magnification of the area delineated by the white rectangle (GFP, A 1 ; DARPP-32, A 2 ; merge A 3 ). Scale bar, 100 μm. Note the absence of D2R-expressing MSNs in a longitudinal stripe bordering the GPe. (B,C) Striatal sections from Drd2-EGFP mice ( n = 5) stained with antibodies against D1R (GFP, B 1 ; D1R, B 2 ; merge, B 3 ) and Gα olf (GFP, C 1 ; DARPP-32, C 2 ; merge C 3 ). Scale bar, 100 μm. (D) Analysis of Ripley's K -function for D1R-MSNs (GFP-negative) and D2R-MSNs (GFP-positive) distributions (green lines) with the average (black line) compared to simulated distributions following CSR (red lines). For the experimental data (green lines) the difference between the estimated K -function and the expected K -function ( ∧ K ( t ) − E [ ∧ K ( t )]) is negative indicating that the D1R- and D2R-MSNs occupy different parts of the sample area; in addition the distribution of the green lines does not overlap with the distribution of the simulated data (red lines) indicating a non-random distribution of D1R-MSNs and D2R-MSNs with respect to each other. Because of the non-random distribution, only the spatial relation between D1R- and D2R-positive MSNs (D1xD2) was evaluated. Inset, schematic illustration of the areas analyzed. (E,F) Pattern of Cre-mediated recombination visualized by immunofluorescence of GFP (green, E and F ) and <t>β-galactosidase</t> (β-Gal; magenta, E ) and DARPP-32 (magenta, F ) in mice carrying both Drd2-Cre and NLS-LacZ-Tau mGFP :LoxP ( n = 3) (E) or RCE:LoxP ( n = 5) transgenes (F) . (G) Pattern of Cre-mediated recombination visualized by immunofluorescence staining using antibodies against GFP (green) and DARPP-32 (magenta) in carrying both Adora2a-Cre and Rosa26:loxP ( n = 5) transgenes. Scale bar, 200 μm. (H) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd1a-EGFP mice ( n = 5). High magnification of the area delineated by the white rectangle (GFP, H 1 ; DARPP-32, H 2 ; merge H 3 ). Scale bar, 100 μm. Note the complete co-localization of DARPP-32 and GFP in a longitudinal stripe bordering the GPe. ( I ) Injection site of AF594-CTB in the SNr. Scale bar, 1 mm. (J) After intra-SNr injection of AF594-CTB retrogradely labeled MSNs are located in the longitudinal stripe bordering the GPe. Scale bar, 500 μm. High magnification of the area delineated by the yellow square (J 1 ) . Scale bar, 100 μm. (K) Injection site for AF594-CTB in the VTA. Scale bar, 1 mm. (L) MSNs located in the area lacking D2R/A2aR-expressing MSNs are not retrogradely labeled after AF594-CTB injection in the VTA. Scale bar, 500 μm. DStr, dorsal striatum; GPe, external globus pallidus; GFP, green fluorescent protein; Th, thalamus; GPi, internal globus pallidus; SNr, substantia nigra pars reticulata; VTA, ventral tegmental area; BlA, basolateral amygdala; CeA, central amygdala.
    Rabbit Anti β Galactosidase, supplied by Valiant, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti β galactosidase/product/Valiant
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti β galactosidase - by Bioz Stars, 2021-05
    92/100 stars
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    91
    Valiant rabbit anti sheep immunoglobulin g
    Podocyte IRE1α deletion result in a reduction of WT1 and synaptopodin. (A–C) Kidney sections from 9-mo-old mice were stained with antibodies to WT1 (A), synaptopodin (B), and podocalyxin (C). Staining with nonimmune <t>IgG</t> is also presented (negative controls). Scale bars, 50 μm. (D, E) The number of WT1-positive nuclei per glomerulus (determined by colocalization with Hoechst nuclear stain; not shown) was assessed by visual counting. WT1 counts per glomerulus were comparable, but when expressed per 1000 µm 2 of glomerular area, WT1 counts were significantly lower in M Cre mice (* p = 4.6 × 10 −5 ); 32 M + glomeruli from three mice and 48 M Cre glomeruli from three mice. (F) Synaptopodin-stained glomeruli were enlarged in M Cre mice (* p = 0.024). (G) Quantification of synaptopodin immunofluorescence per unit glomerular area showed that intensity was lower in M Cre mice (* p = 0.046). For F and G, 74 M + glomeruli from three mice and 69 M Cre glomeruli from three mice. (H) Podocalyxin-stained glomeruli were enlarged in M Cre mice (* p = 0.0013). (I) Podocalyxin fluorescence per glomerulus and fluorescence per unit glomerular area were comparable in both groups. For H and I, 58 M + glomeruli from three mice and 54 M Cre glomeruli from three mice.
    Rabbit Anti Sheep Immunoglobulin G, supplied by Valiant, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti sheep immunoglobulin g/product/Valiant
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti sheep immunoglobulin g - by Bioz Stars, 2021-05
    91/100 stars
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    N/A
    Product is horseradish peroxidase HRP conjugated rabbit affinity purified antibody to mouse IgG whole molecule and buffer salts
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    Image Search Results


    Segregated distribution of D1R- and D2R/A2aR-expressing MSNs in the caudal part of the striatum . (A) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd2-EGFP mice ( n = 10). Scale bar, 200 μm. High magnification of the area delineated by the white rectangle (GFP, A 1 ; DARPP-32, A 2 ; merge A 3 ). Scale bar, 100 μm. Note the absence of D2R-expressing MSNs in a longitudinal stripe bordering the GPe. (B,C) Striatal sections from Drd2-EGFP mice ( n = 5) stained with antibodies against D1R (GFP, B 1 ; D1R, B 2 ; merge, B 3 ) and Gα olf (GFP, C 1 ; DARPP-32, C 2 ; merge C 3 ). Scale bar, 100 μm. (D) Analysis of Ripley's K -function for D1R-MSNs (GFP-negative) and D2R-MSNs (GFP-positive) distributions (green lines) with the average (black line) compared to simulated distributions following CSR (red lines). For the experimental data (green lines) the difference between the estimated K -function and the expected K -function ( ∧ K ( t ) − E [ ∧ K ( t )]) is negative indicating that the D1R- and D2R-MSNs occupy different parts of the sample area; in addition the distribution of the green lines does not overlap with the distribution of the simulated data (red lines) indicating a non-random distribution of D1R-MSNs and D2R-MSNs with respect to each other. Because of the non-random distribution, only the spatial relation between D1R- and D2R-positive MSNs (D1xD2) was evaluated. Inset, schematic illustration of the areas analyzed. (E,F) Pattern of Cre-mediated recombination visualized by immunofluorescence of GFP (green, E and F ) and β-galactosidase (β-Gal; magenta, E ) and DARPP-32 (magenta, F ) in mice carrying both Drd2-Cre and NLS-LacZ-Tau mGFP :LoxP ( n = 3) (E) or RCE:LoxP ( n = 5) transgenes (F) . (G) Pattern of Cre-mediated recombination visualized by immunofluorescence staining using antibodies against GFP (green) and DARPP-32 (magenta) in carrying both Adora2a-Cre and Rosa26:loxP ( n = 5) transgenes. Scale bar, 200 μm. (H) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd1a-EGFP mice ( n = 5). High magnification of the area delineated by the white rectangle (GFP, H 1 ; DARPP-32, H 2 ; merge H 3 ). Scale bar, 100 μm. Note the complete co-localization of DARPP-32 and GFP in a longitudinal stripe bordering the GPe. ( I ) Injection site of AF594-CTB in the SNr. Scale bar, 1 mm. (J) After intra-SNr injection of AF594-CTB retrogradely labeled MSNs are located in the longitudinal stripe bordering the GPe. Scale bar, 500 μm. High magnification of the area delineated by the yellow square (J 1 ) . Scale bar, 100 μm. (K) Injection site for AF594-CTB in the VTA. Scale bar, 1 mm. (L) MSNs located in the area lacking D2R/A2aR-expressing MSNs are not retrogradely labeled after AF594-CTB injection in the VTA. Scale bar, 500 μm. DStr, dorsal striatum; GPe, external globus pallidus; GFP, green fluorescent protein; Th, thalamus; GPi, internal globus pallidus; SNr, substantia nigra pars reticulata; VTA, ventral tegmental area; BlA, basolateral amygdala; CeA, central amygdala.

    Journal: Frontiers in Neural Circuits

    Article Title: Spatial distribution of D1R- and D2R-expressing medium-sized spiny neurons differs along the rostro-caudal axis of the mouse dorsal striatum

    doi: 10.3389/fncir.2013.00124

    Figure Lengend Snippet: Segregated distribution of D1R- and D2R/A2aR-expressing MSNs in the caudal part of the striatum . (A) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd2-EGFP mice ( n = 10). Scale bar, 200 μm. High magnification of the area delineated by the white rectangle (GFP, A 1 ; DARPP-32, A 2 ; merge A 3 ). Scale bar, 100 μm. Note the absence of D2R-expressing MSNs in a longitudinal stripe bordering the GPe. (B,C) Striatal sections from Drd2-EGFP mice ( n = 5) stained with antibodies against D1R (GFP, B 1 ; D1R, B 2 ; merge, B 3 ) and Gα olf (GFP, C 1 ; DARPP-32, C 2 ; merge C 3 ). Scale bar, 100 μm. (D) Analysis of Ripley's K -function for D1R-MSNs (GFP-negative) and D2R-MSNs (GFP-positive) distributions (green lines) with the average (black line) compared to simulated distributions following CSR (red lines). For the experimental data (green lines) the difference between the estimated K -function and the expected K -function ( ∧ K ( t ) − E [ ∧ K ( t )]) is negative indicating that the D1R- and D2R-MSNs occupy different parts of the sample area; in addition the distribution of the green lines does not overlap with the distribution of the simulated data (red lines) indicating a non-random distribution of D1R-MSNs and D2R-MSNs with respect to each other. Because of the non-random distribution, only the spatial relation between D1R- and D2R-positive MSNs (D1xD2) was evaluated. Inset, schematic illustration of the areas analyzed. (E,F) Pattern of Cre-mediated recombination visualized by immunofluorescence of GFP (green, E and F ) and β-galactosidase (β-Gal; magenta, E ) and DARPP-32 (magenta, F ) in mice carrying both Drd2-Cre and NLS-LacZ-Tau mGFP :LoxP ( n = 3) (E) or RCE:LoxP ( n = 5) transgenes (F) . (G) Pattern of Cre-mediated recombination visualized by immunofluorescence staining using antibodies against GFP (green) and DARPP-32 (magenta) in carrying both Adora2a-Cre and Rosa26:loxP ( n = 5) transgenes. Scale bar, 200 μm. (H) DARPP-32 (magenta) and GFP (green) immunofluorescence in the caudal part of the dorsal striatum of Drd1a-EGFP mice ( n = 5). High magnification of the area delineated by the white rectangle (GFP, H 1 ; DARPP-32, H 2 ; merge H 3 ). Scale bar, 100 μm. Note the complete co-localization of DARPP-32 and GFP in a longitudinal stripe bordering the GPe. ( I ) Injection site of AF594-CTB in the SNr. Scale bar, 1 mm. (J) After intra-SNr injection of AF594-CTB retrogradely labeled MSNs are located in the longitudinal stripe bordering the GPe. Scale bar, 500 μm. High magnification of the area delineated by the yellow square (J 1 ) . Scale bar, 100 μm. (K) Injection site for AF594-CTB in the VTA. Scale bar, 1 mm. (L) MSNs located in the area lacking D2R/A2aR-expressing MSNs are not retrogradely labeled after AF594-CTB injection in the VTA. Scale bar, 500 μm. DStr, dorsal striatum; GPe, external globus pallidus; GFP, green fluorescent protein; Th, thalamus; GPi, internal globus pallidus; SNr, substantia nigra pars reticulata; VTA, ventral tegmental area; BlA, basolateral amygdala; CeA, central amygdala.

    Article Snippet: Finally, they were incubated overnight or 72 h at 4°C with the primary antibodies: chicken and rabbit anti-GFP (1:500 and 1:1000 respectively, Invitrogen), rabbit anti-vesicular glutamate transporter 1 (VGluT1) or anti-VGluT2 (1:1000 gift from S. El Mestikawy), mouse anti-tyrosine hydroxylase (TH) (1:1000, Millipore), rat anti-dopamine transporter (DAT) (1:1000, Millipore), mouse anti-NeuN (1:500, Millipore), mouse anti-D1R (1:500 gift from R. R. Luedtke), rabbit anti-Gα olf (1:500) (Hervé et al., ), rabbit anti-β-galactosidase (1:1000, Cappel, MP Biomedicals), guinea-pig anti-MOR (1:500 gift from T. Kaneko) mouse anti-DARPP-32 (1:1000 gift from P. Greengard), rabbit anti-calretinin (CalR), anti-calbindin-D28k and anti-parvalbumin (ParV) (1:1000, Swant), rabbit anti-neuropeptide Y (NPY) (1:400, Abcam), goat anti-ChAT (1:400, Millipore), rabbit anti-substance P (1:500, Millipore), rabbit anti-preproenkephalin (ppENK) (1:500, Neuromics) and rabbit anti-RFP (1:1000, MBL).

    Techniques: Expressing, Immunofluorescence, Mouse Assay, Staining, Injection, CtB Assay, Labeling

    Comparative Dll4 expression in postnatal and adult retinas. (A1-B3) β-gal activity in P1 postnatal retinas from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. Both lines are clearly active in non-endothelial cell types within the retina at this stage. (C1-D3) β-gal activity in P5 postnatal retinas from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. Dll4-BAC-nlacZ is more active than the endogenous reporter in the postnatal retinal arterial endothelium. (E1-F3) β-gal activity is detected in the arterial and capillary endothelium in both (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ retinas at P7. (G1-H3) Dll4 lacZ/+ is enriched in arteries and capillary vessels (and weakly detected in veins), while Dll4-BAC-nlacZ has no activity within the veins in the adult retina. (I1-I6) IHC and indirect immunofluorescent detection of isolectin (I1), smooth muscle actin (I2), β-gal (I3), and merged (I4-I6) images from representative Dll4-BAC-nlacZ adult retinas. a, artery; on, optic nerve; v, vein. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression in postnatal and adult retinas. (A1-B3) β-gal activity in P1 postnatal retinas from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. Both lines are clearly active in non-endothelial cell types within the retina at this stage. (C1-D3) β-gal activity in P5 postnatal retinas from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. Dll4-BAC-nlacZ is more active than the endogenous reporter in the postnatal retinal arterial endothelium. (E1-F3) β-gal activity is detected in the arterial and capillary endothelium in both (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ retinas at P7. (G1-H3) Dll4 lacZ/+ is enriched in arteries and capillary vessels (and weakly detected in veins), while Dll4-BAC-nlacZ has no activity within the veins in the adult retina. (I1-I6) IHC and indirect immunofluorescent detection of isolectin (I1), smooth muscle actin (I2), β-gal (I3), and merged (I4-I6) images from representative Dll4-BAC-nlacZ adult retinas. a, artery; on, optic nerve; v, vein. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Mouse Assay, Immunohistochemistry

    Comparative Dll4 expression during intermediate and late-stage embryonic development. (A-C) lacZ activity in E12.5 (A), E14.5 (B), and E18.5 (C) Dll4 lacZ/+ mouse embryos and yolk sacs. (D) Intra-littermate body measurements in a Dll4 lacZ/+ litter. Data are presented as averages ±s.e.m. Comparisons were made by Student's t -test (** P =0.0058). (E-G″) lacZ activity in E12.5, E14.5, and E18.5 Dll4-BAC-nlacZ mouse embryos (E-G) and yolk sacs (E′-G″). (H) Intra-littermate body measurements in a Dll4-BAC-nlacZ litter. Data are presented as averages ±s.e.m.; ns, nonsignificant. Comparisons were made by Student's t -test. Noticeable size differences can be observed between genotypes due to heterozygous Dll4 loss of function. (I-J) β-gal IHC on E14.5 skin from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ embryos. I′ and J′ are magnified views of a respective region shown in corresponding panels I and J. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression during intermediate and late-stage embryonic development. (A-C) lacZ activity in E12.5 (A), E14.5 (B), and E18.5 (C) Dll4 lacZ/+ mouse embryos and yolk sacs. (D) Intra-littermate body measurements in a Dll4 lacZ/+ litter. Data are presented as averages ±s.e.m. Comparisons were made by Student's t -test (** P =0.0058). (E-G″) lacZ activity in E12.5, E14.5, and E18.5 Dll4-BAC-nlacZ mouse embryos (E-G) and yolk sacs (E′-G″). (H) Intra-littermate body measurements in a Dll4-BAC-nlacZ litter. Data are presented as averages ±s.e.m.; ns, nonsignificant. Comparisons were made by Student's t -test. Noticeable size differences can be observed between genotypes due to heterozygous Dll4 loss of function. (I-J) β-gal IHC on E14.5 skin from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ embryos. I′ and J′ are magnified views of a respective region shown in corresponding panels I and J. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Immunohistochemistry

    β-gal reporter activity is restricted to arterial vasculature in the skin. (A1-A6) Single channel views of indirect immunofluorescence for CD31 (A1), Dll4 (A2), β-gal (A3), and merged (A4-A6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4 lacZ/+ mouse skin. (B1-B6) CD31 (B1), Podoplanin (B2), β-gal (B3), and merged (B4-B6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4 lacZ/+ mouse skin. (C1-C6) CD31 (C1), SMA (C2), β-gal (C3), and merged (C4-C6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker, smooth muscle actin (SMA). (D1-D6) CD31 (D1), Tuj1 (D2), β-gal (D3), and merged (D4-D6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. (E1-E6) CD31 (E1), Dll4 (E2), β-gal (E3), and merged (E4-E6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4-BAC-nlacZ mouse skin. (F1-F6) CD31 (F1), Podoplanin (F2), β-gal (F3), and merged (F4-F6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4-BAC-nlacZ mouse skin. (G1-G6) CD31 (G1), SMA (G2), β-gal (G3), and merged (G4-G6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker SMA. (H1-H6) CD31 (H1), Tuj1 (H2), β-gal (H3), and merged (H4-H6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: β-gal reporter activity is restricted to arterial vasculature in the skin. (A1-A6) Single channel views of indirect immunofluorescence for CD31 (A1), Dll4 (A2), β-gal (A3), and merged (A4-A6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4 lacZ/+ mouse skin. (B1-B6) CD31 (B1), Podoplanin (B2), β-gal (B3), and merged (B4-B6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4 lacZ/+ mouse skin. (C1-C6) CD31 (C1), SMA (C2), β-gal (C3), and merged (C4-C6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker, smooth muscle actin (SMA). (D1-D6) CD31 (D1), Tuj1 (D2), β-gal (D3), and merged (D4-D6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. (E1-E6) CD31 (E1), Dll4 (E2), β-gal (E3), and merged (E4-E6) images showing colocalization between β-gal-positive vasculature and endogenous Dll4 in Dll4-BAC-nlacZ mouse skin. (F1-F6) CD31 (F1), Podoplanin (F2), β-gal (F3), and merged (F4-F6) images showing a lack of colocalization between β-gal-positive vasculature and the lymphatic-specific marker Podoplanin in Dll4-BAC-nlacZ mouse skin. (G1-G6) CD31 (G1), SMA (G2), β-gal (G3), and merged (G4-G6) images showing colocalization between β-gal-positive vasculature and the arterial-specific marker SMA. (H1-H6) CD31 (H1), Tuj1 (H2), β-gal (H3), and merged (H4-H6) images showing lack of colocalization between β-gal-positive vasculature and the neuronal-specific marker Tuj1. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Activity Assay, Immunofluorescence, Marker, BAC Assay

    Comparative Dll4 expression in intermediate and late-stage embryonic hearts and lungs. (A1-B5) β-gal activity in E12.5 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5. β-gal activity is present within coronary plexus (A1,A2,B1,B2), the endocardium of the distal end (A1,B1) and root of the aorta (A4,B4) and pulmonary artery in both lines, as well as the endocardium and subepicardial vasculature (A5,B5), but absent from the epicardium and myocardium. (C1-D5) β-gal activity in E14.5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5. β-gal activity is localized to the endocardium of the aorta in both lines (C4,D4), as well as the chamber endocardium (C5,D5), and subepicardial coronary vasculature (C5,D5). β-gal activity was also detected within a small fraction of the myocardium in the BAC reporter line at this stage. (E1-F5) β-gal activity in E18.5 hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root in Dll4 lacZ/+ animals, but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5), and sparsely in the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle. (G1-H4) β-gal activity in E12.5 lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 are representative cross-sections through the lungs, and boxed in areas are magnified in G4 and H4, revealing activity within the endothelium. (I1-J4) β-gal activity in E14.5 lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-J2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4, revealing endothelial-specific activity in both lines. (K1-L4) β-gal activity in E18.5 lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4, demonstrating endothelial-specific activity in both lines. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression in intermediate and late-stage embryonic hearts and lungs. (A1-B5) β-gal activity in E12.5 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5. β-gal activity is present within coronary plexus (A1,A2,B1,B2), the endocardium of the distal end (A1,B1) and root of the aorta (A4,B4) and pulmonary artery in both lines, as well as the endocardium and subepicardial vasculature (A5,B5), but absent from the epicardium and myocardium. (C1-D5) β-gal activity in E14.5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5. β-gal activity is localized to the endocardium of the aorta in both lines (C4,D4), as well as the chamber endocardium (C5,D5), and subepicardial coronary vasculature (C5,D5). β-gal activity was also detected within a small fraction of the myocardium in the BAC reporter line at this stage. (E1-F5) β-gal activity in E18.5 hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root in Dll4 lacZ/+ animals, but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5), and sparsely in the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle. (G1-H4) β-gal activity in E12.5 lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 are representative cross-sections through the lungs, and boxed in areas are magnified in G4 and H4, revealing activity within the endothelium. (I1-J4) β-gal activity in E14.5 lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-J2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4, revealing endothelial-specific activity in both lines. (K1-L4) β-gal activity in E18.5 lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4, demonstrating endothelial-specific activity in both lines. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression in intermediate and late-stage wholemount embryonic brains. (A1-B9) β-gal activity in the E12.5 embryonic brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the E14.5 embryonic brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the E18.5 embryonic brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; AIC, anterior inferior cerebellar artery; azACA, azygos of the anterior cerebral artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression in intermediate and late-stage wholemount embryonic brains. (A1-B9) β-gal activity in the E12.5 embryonic brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the E14.5 embryonic brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the E18.5 embryonic brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; AIC, anterior inferior cerebellar artery; azACA, azygos of the anterior cerebral artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression during early embryonic development. (A) Schematic of the BAC transgene used for generating the Dll4-BAC-nlacZ mouse line, with a magnified schematic of the nuclear LacZ insertion at the ATG start site of Dll4 . (B-E) β-gal activity in E7.75-E10.5 Dll4 lacZ/+ mouse embryos, ventral (B,C) and sagittal (D,E) views. (F-I) β-gal activity in E7.75-E10.5 Dll4-BAC-nlacZ mouse embryos, ventral (F,G) and sagittal (H,I) views. E′ and I′ show β-gal activity in the embryonic yolk sac. E″ and I″ are magnified views of a representative region shown in corresponding panels E′ and I′, respectively. (J-M) Coronal view of X-gal-stained and Eosin-counterstained sections of E9.5 and E10.5 Dll4 lacZ/+ and Dll4-BAC-nlacZ embryos. aPCs, aortic progenitor cells; CC, cardiac crescent; CV, cardinal vein; DA, dorsal aorta; EC, endocardium; HB, hindbrain; ICA, internal carotid artery; ISA, intersegmental artery; LA, left atrium; LV, left ventricle; NT, neural tube; OFT, outflow tract; PNVP, perineural vascular plexus; RA, right atrium; RV, right ventricle; SV, sinus venosus; black caret, ventral V2 interneuron population. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression during early embryonic development. (A) Schematic of the BAC transgene used for generating the Dll4-BAC-nlacZ mouse line, with a magnified schematic of the nuclear LacZ insertion at the ATG start site of Dll4 . (B-E) β-gal activity in E7.75-E10.5 Dll4 lacZ/+ mouse embryos, ventral (B,C) and sagittal (D,E) views. (F-I) β-gal activity in E7.75-E10.5 Dll4-BAC-nlacZ mouse embryos, ventral (F,G) and sagittal (H,I) views. E′ and I′ show β-gal activity in the embryonic yolk sac. E″ and I″ are magnified views of a representative region shown in corresponding panels E′ and I′, respectively. (J-M) Coronal view of X-gal-stained and Eosin-counterstained sections of E9.5 and E10.5 Dll4 lacZ/+ and Dll4-BAC-nlacZ embryos. aPCs, aortic progenitor cells; CC, cardiac crescent; CV, cardinal vein; DA, dorsal aorta; EC, endocardium; HB, hindbrain; ICA, internal carotid artery; ISA, intersegmental artery; LA, left atrium; LV, left ventricle; NT, neural tube; OFT, outflow tract; PNVP, perineural vascular plexus; RA, right atrium; RV, right ventricle; SV, sinus venosus; black caret, ventral V2 interneuron population. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, BAC Assay, Activity Assay, Staining

    Comparative Dll4 expression in wholemount postnatal and adult brains. (A1-B3) β-gal activity in the P1 postnatal brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the P5 postnatal brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the adult brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; azACA, azygos of the anterior cerebral artery; AIC, anterior inferior cerebellar artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression in wholemount postnatal and adult brains. (A1-B3) β-gal activity in the P1 postnatal brain of (A) Dll4 lacZ/+ and (B) Dll4-BAC-nlacZ mice. A1-A3 and B1-B3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. A4-A9 and B4-B9 show representative coronal sections through the brain, from anterior to posterior. (C1-D9) β-gal activity in the P5 postnatal brain of (C) Dll4 lacZ/+ and (D) Dll4-BAC-nlacZ mice. C1-C3 and D1-D3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. C4-C9 and D4-D9 show representative coronal sections through the brain, from anterior to posterior. (E1-F9) β-gal activity in the adult brain of (E) Dll4 lacZ/+ and (F) Dll4-BAC-nlacZ mice. E1-E3 and F1-F3 show representative wholemount images of the brain from superior, inferior, and sagittal planes. E4-E9 and F4-F9 show representative coronal sections through the brain, from anterior to posterior. ACA, anterior cerebral artery; azACA, azygos of the anterior cerebral artery; AIC, anterior inferior cerebellar artery; BA, basilar artery; ICA, internal carotid artery; MCA, middle cerebral artery; PCA, posterior cerebral artery; SCA, superior cerebellar artery; VA, vertebral artery. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Comparative Dll4 expression in postnatal and adult hearts and lungs. (A1-B5) β-gal activity in P1 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5, with activity evident within the endocardial lining of the aorta in both lines (A4,B4), as well as the endocardium, coronary vasculature, and myocardium (A5,B5). (C1-D5) β-gal activity in P5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5, with signal evident within the endocardial lining of the aorta in both lines, and persisting in the endocardium of the aorta (C4,D4) and chambers, as well as the myocardium and coronary vasculature (C5,D5). (E1-F5) β-gal activity in adult hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from adult Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, magnified in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root Dll4 lacZ/+ but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5) (asterisks), and within the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle; sm, smooth muscle; asterisks – denote lumenized vasculature. (G1-H4) β-gal activity in P1 postnatal lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels G4 and H4. (I1-J4) β-gal activity in P5 postnatal lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-D2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4. (K1-L4) β-gal activity in adult lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4. In both lines, and at all stages, β-gal activity appears to be confined to the endothelium. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Journal: Biology Open

    Article Title: A novel reporter allele for monitoring Dll4 expression within the embryonic and adult mouse

    doi: 10.1242/bio.026799

    Figure Lengend Snippet: Comparative Dll4 expression in postnatal and adult hearts and lungs. (A1-B5) β-gal activity in P1 hearts from (A) Dll4 lacZ/+ or (B) Dll4-BAC-nlacZ mice. A1-A2 and B1-B2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. A3 and B3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels A4-A5 and B4-B5, with activity evident within the endocardial lining of the aorta in both lines (A4,B4), as well as the endocardium, coronary vasculature, and myocardium (A5,B5). (C1-D5) β-gal activity in P5 hearts from (C) Dll4 lacZ/+ or (D) Dll4-BAC-nlacZ mice. C1-C2 and D1-D2 show representative wholemount hearts from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. C3 and D3 show β-gal activity in a representative cross-section through the heart, which is magnified accordingly in panels C4-C5 and D4-D5, with signal evident within the endocardial lining of the aorta in both lines, and persisting in the endocardium of the aorta (C4,D4) and chambers, as well as the myocardium and coronary vasculature (C5,D5). (E1-F5) β-gal activity in adult hearts from (E) Dll4 lacZ/+ or (F) Dll4-BAC-nlacZ mice. E1-E2 and F1-F2 show representative wholemount hearts from adult Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. E3 and F3 show β-gal activity in a representative cross-section through the heart, magnified in panels E4-E5 and F4-F5. β-gal activity is localized to the endocardium of the aortic root Dll4 lacZ/+ but absent from Dll4-BAC-nlacZ mice (E4,F4), and present in both lines within the chamber endocardium and coronary vasculature (E5,F5) (asterisks), and within the myocardium. Ao, aorta; ec, endocardium; ep, epicardium; IVS, interventricular septum; LA, left atrium; LV, left ventricle; m, myocardium; PA, pulmonary artery; RA, right atrium; RV, right ventricle; sm, smooth muscle; asterisks – denote lumenized vasculature. (G1-H4) β-gal activity in P1 postnatal lungs from (G) Dll4 lacZ/+ or (H) Dll4-BAC-nlacZ mice. G1-G2 and H1-H2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. G3 and H3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels G4 and H4. (I1-J4) β-gal activity in P5 postnatal lungs from (I) Dll4 lacZ/+ or (J) Dll4-BAC-nlacZ mice. I1-I2 and J1-D2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. I3 and J3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels I4 and J4. (K1-L4) β-gal activity in adult lungs from (K) Dll4 lacZ/+ or (L) Dll4-BAC-nlacZ mice. K1-K2 and L1-L2 show representative wholemount lungs from Dll4 lacZ/+ and Dll4-BAC-nlacZ mice, respectively, from ventral and dorsal views. K3 and L3 show β-gal activity in a representative cross-section through the lungs, which is magnified accordingly in panels K4 and L4. In both lines, and at all stages, β-gal activity appears to be confined to the endothelium. D, dorsal; e, endothelium; L, left; R, right; sm, smooth muscle; V, ventral. Units depicted are in μm.

    Article Snippet: Primary antibodies used were: goat anti-Dll4 (1:100) (R & D Systems, AF1389), rat anti-CD31 (1:200) (BD Pharmingen, San Jose, USA, 550274), hamster anti-Podoplanin (1:100) (Developmental Studies Hybridoma Bank, 8.1.1), mouse anti-Tuj1 (1:500) (Covance, Princeton, USA, MMS-435P), rabbit anti-β-galactosidase (1:1000) (MP Biomedical, Santa Ana, USA, 55976), biotinylated Griffonia Simplicifolia Lectin I isolectin B4 (1:50) (Vector Laboratories, B-1205), and mouse anti-actin, alpha-smooth muscle FITC (1:100) (Sigma-Aldrich, F3777).

    Techniques: Expressing, Activity Assay, BAC Assay, Mouse Assay

    Analysis of Plexin-B2 expression in sensory neurons of the dorsal root ganglia (DRG) in adult mice and its regulation in inflammatory pain. a , b Expression of plxnb2 via immunofluorescence analysis of β-galactosidase in adult DRG in respective LacZ reporter knock-in mice. Typical examples a and quantitative analysis b of the distribution of DRG cell types expressing plxnb2 via co-immunolabeling with marker proteins ( n = 10–20 sections/group taken from at least three different mice/group). Scale bar , 50 µm. c , d Typical examples c and quantitative summary d of LacZ staining demonstrating plxnb2 expression in adult Plexin-B2-LacZ +/− mice following intraplantar injection of either vehicle (control) or Complete Freund´s Adjuvant (CFA) stimulating inflammatory pain ( n = 20–30 sections/group taken from at least 3 different mice/group). Scale bars represent 50 µm. In d , Student’s t -test (two sides) was performed. P

    Journal: Nature Communications

    Article Title: Semaphorin 4C Plexin-B2 signaling in peripheral sensory neurons is pronociceptive in a model of inflammatory pain

    doi: 10.1038/s41467-017-00341-w

    Figure Lengend Snippet: Analysis of Plexin-B2 expression in sensory neurons of the dorsal root ganglia (DRG) in adult mice and its regulation in inflammatory pain. a , b Expression of plxnb2 via immunofluorescence analysis of β-galactosidase in adult DRG in respective LacZ reporter knock-in mice. Typical examples a and quantitative analysis b of the distribution of DRG cell types expressing plxnb2 via co-immunolabeling with marker proteins ( n = 10–20 sections/group taken from at least three different mice/group). Scale bar , 50 µm. c , d Typical examples c and quantitative summary d of LacZ staining demonstrating plxnb2 expression in adult Plexin-B2-LacZ +/− mice following intraplantar injection of either vehicle (control) or Complete Freund´s Adjuvant (CFA) stimulating inflammatory pain ( n = 20–30 sections/group taken from at least 3 different mice/group). Scale bars represent 50 µm. In d , Student’s t -test (two sides) was performed. P

    Article Snippet: DRGs were stained with anti-β-galactosidase antibodies (rabbit 1:700; MP Biomedicals, 559762 and chicken 1:800; Abcam, ab9361), anti-PlexinB2 (rabbit 1:25, CSB-PA347467, CusAb-Cusabio), anti-CGRP antibody (1:2000; 24112, Immunostar), biotinylated Isolectin B4 (1:200; B-1205, Vector), and anti-NF200 antibody (1:300, N4142, Sigma).

    Techniques: Expressing, Mouse Assay, Immunofluorescence, Knock-In, Immunolabeling, Marker, Staining, Injection

    Sema4C is expressed in adult DRG and paw skin and plays a functional role in inflammatory pain. a Expression of Sema4c via β-galactosidase (LacZ) staining in adult DRG and plantar paw skin using LacZ reporter knock-in mice. Typical examples ( upper ) and quantitative summary (panel) of LacZ staining in the basal state or following CFA-induced paw inflammation. Scale bars , 50 µm. b Expression of Sema4C and its co-localization with immune cells markers via double immunofluorescence staining in plantar paw skin of mice at 24 h after vehicle or CFA injection, using anti-Sema4C antibody and antibodies against immune cells (CD3 to mark infiltrating T cells, upper ; or GR-1 to target macrophages, lower ). Higher magnification views of dermis are shown on extreme right to illustrate cells showing co-localization ( arrowheads ). Scale bars , 25 µm. c Quantitative measurement of intensity of Sema4C immunoreactivity paw tissue 24 h after intraplantar injection of vehicle or CFA; n = 3 mice/group. d , e Examples ( left ) and densitometric quantifications ( right ) of western blot analysis of Sema4C signal in lysates of L3-L4 DRGs d or paw tissue e 24 h after intraplantar CFA injection; n = 8 for DRGs, n = 9 for paw tissues. f Analysis of inflammatory mechanical hypersensitivity following hindpaw CFA injection in mice lacking Sema4C (Sema4C −/− ) and their wild-type littermates. Frequency of paw withdrawal in response to application of 0.07 g force via a von Frey filament is shown. g Changes in paw response latency to radiant heat following CFA injection in Sema4C −/− mice and their wild-type controls littermates. n = 5 (Sema4C −/− mice) and n = 7 (wild-type littermates) mice/group. Student’s t -test was performed in a – e and two-way ANOVA for repeated measures followed by Tukey’s test was performed in f and g . In f , P

    Journal: Nature Communications

    Article Title: Semaphorin 4C Plexin-B2 signaling in peripheral sensory neurons is pronociceptive in a model of inflammatory pain

    doi: 10.1038/s41467-017-00341-w

    Figure Lengend Snippet: Sema4C is expressed in adult DRG and paw skin and plays a functional role in inflammatory pain. a Expression of Sema4c via β-galactosidase (LacZ) staining in adult DRG and plantar paw skin using LacZ reporter knock-in mice. Typical examples ( upper ) and quantitative summary (panel) of LacZ staining in the basal state or following CFA-induced paw inflammation. Scale bars , 50 µm. b Expression of Sema4C and its co-localization with immune cells markers via double immunofluorescence staining in plantar paw skin of mice at 24 h after vehicle or CFA injection, using anti-Sema4C antibody and antibodies against immune cells (CD3 to mark infiltrating T cells, upper ; or GR-1 to target macrophages, lower ). Higher magnification views of dermis are shown on extreme right to illustrate cells showing co-localization ( arrowheads ). Scale bars , 25 µm. c Quantitative measurement of intensity of Sema4C immunoreactivity paw tissue 24 h after intraplantar injection of vehicle or CFA; n = 3 mice/group. d , e Examples ( left ) and densitometric quantifications ( right ) of western blot analysis of Sema4C signal in lysates of L3-L4 DRGs d or paw tissue e 24 h after intraplantar CFA injection; n = 8 for DRGs, n = 9 for paw tissues. f Analysis of inflammatory mechanical hypersensitivity following hindpaw CFA injection in mice lacking Sema4C (Sema4C −/− ) and their wild-type littermates. Frequency of paw withdrawal in response to application of 0.07 g force via a von Frey filament is shown. g Changes in paw response latency to radiant heat following CFA injection in Sema4C −/− mice and their wild-type controls littermates. n = 5 (Sema4C −/− mice) and n = 7 (wild-type littermates) mice/group. Student’s t -test was performed in a – e and two-way ANOVA for repeated measures followed by Tukey’s test was performed in f and g . In f , P

    Article Snippet: DRGs were stained with anti-β-galactosidase antibodies (rabbit 1:700; MP Biomedicals, 559762 and chicken 1:800; Abcam, ab9361), anti-PlexinB2 (rabbit 1:25, CSB-PA347467, CusAb-Cusabio), anti-CGRP antibody (1:2000; 24112, Immunostar), biotinylated Isolectin B4 (1:200; B-1205, Vector), and anti-NF200 antibody (1:300, N4142, Sigma).

    Techniques: Functional Assay, Expressing, Staining, Knock-In, Mouse Assay, Double Immunofluorescence Staining, Injection, Western Blot

    Podocyte IRE1α deletion result in a reduction of WT1 and synaptopodin. (A–C) Kidney sections from 9-mo-old mice were stained with antibodies to WT1 (A), synaptopodin (B), and podocalyxin (C). Staining with nonimmune IgG is also presented (negative controls). Scale bars, 50 μm. (D, E) The number of WT1-positive nuclei per glomerulus (determined by colocalization with Hoechst nuclear stain; not shown) was assessed by visual counting. WT1 counts per glomerulus were comparable, but when expressed per 1000 µm 2 of glomerular area, WT1 counts were significantly lower in M Cre mice (* p = 4.6 × 10 −5 ); 32 M + glomeruli from three mice and 48 M Cre glomeruli from three mice. (F) Synaptopodin-stained glomeruli were enlarged in M Cre mice (* p = 0.024). (G) Quantification of synaptopodin immunofluorescence per unit glomerular area showed that intensity was lower in M Cre mice (* p = 0.046). For F and G, 74 M + glomeruli from three mice and 69 M Cre glomeruli from three mice. (H) Podocalyxin-stained glomeruli were enlarged in M Cre mice (* p = 0.0013). (I) Podocalyxin fluorescence per glomerulus and fluorescence per unit glomerular area were comparable in both groups. For H and I, 58 M + glomeruli from three mice and 54 M Cre glomeruli from three mice.

    Journal: Molecular Biology of the Cell

    Article Title: Deletion of inositol-requiring enzyme-1α in podocytes disrupts glomerular capillary integrity and autophagy

    doi: 10.1091/mbc.E16-12-0828

    Figure Lengend Snippet: Podocyte IRE1α deletion result in a reduction of WT1 and synaptopodin. (A–C) Kidney sections from 9-mo-old mice were stained with antibodies to WT1 (A), synaptopodin (B), and podocalyxin (C). Staining with nonimmune IgG is also presented (negative controls). Scale bars, 50 μm. (D, E) The number of WT1-positive nuclei per glomerulus (determined by colocalization with Hoechst nuclear stain; not shown) was assessed by visual counting. WT1 counts per glomerulus were comparable, but when expressed per 1000 µm 2 of glomerular area, WT1 counts were significantly lower in M Cre mice (* p = 4.6 × 10 −5 ); 32 M + glomeruli from three mice and 48 M Cre glomeruli from three mice. (F) Synaptopodin-stained glomeruli were enlarged in M Cre mice (* p = 0.024). (G) Quantification of synaptopodin immunofluorescence per unit glomerular area showed that intensity was lower in M Cre mice (* p = 0.046). For F and G, 74 M + glomeruli from three mice and 69 M Cre glomeruli from three mice. (H) Podocalyxin-stained glomeruli were enlarged in M Cre mice (* p = 0.0013). (I) Podocalyxin fluorescence per glomerulus and fluorescence per unit glomerular area were comparable in both groups. For H and I, 58 M + glomeruli from three mice and 54 M Cre glomeruli from three mice.

    Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse complement C3 antibody (Cappel 55500) and rabbit anti-sheep immunoglobulin G (IgG; 0865205) were purchased from MP Biomedicals (Santa Ana, CA).

    Techniques: Mouse Assay, Staining, Immunofluorescence, Fluorescence