pv immunostaining  (Vector Laboratories)


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    Name:
    Unconjugated Anti Streptavidin Antibody
    Description:
    Anti Streptavidin Antibody has been widely used as amplifying reagents in immunohistochemistry in situ hybridization microarray assays ELISAs blots and many other applications Our antibodies to streptavidin are produced in goats using our highly purified streptavidin and isolated by affinity chromatography Anti Streptavidin does not bind avidin and Anti Avidin does not recognize streptavidin These antibodies provide opportunities to significantly amplify signals in many applications
    Catalog Number:
    sp-4000
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    None
    Host:
    Goat
    Size:
    1 mg
    Category:
    Antibodies
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    Structured Review

    Vector Laboratories pv immunostaining
    Unconjugated Anti Streptavidin Antibody
    Anti Streptavidin Antibody has been widely used as amplifying reagents in immunohistochemistry in situ hybridization microarray assays ELISAs blots and many other applications Our antibodies to streptavidin are produced in goats using our highly purified streptavidin and isolated by affinity chromatography Anti Streptavidin does not bind avidin and Anti Avidin does not recognize streptavidin These antibodies provide opportunities to significantly amplify signals in many applications
    https://www.bioz.com/result/pv immunostaining/product/Vector Laboratories
    Average 85 stars, based on 21579 article reviews
    Price from $9.99 to $1999.99
    pv immunostaining - by Bioz Stars, 2021-02
    85/100 stars

    Images

    1) Product Images from "Pericellular Innervation of Neurons Expressing Abnormally Hyperphosphorylated Tau in the Hippocampal Formation of Alzheimer's Disease Patients"

    Article Title: Pericellular Innervation of Neurons Expressing Abnormally Hyperphosphorylated Tau in the Hippocampal Formation of Alzheimer's Disease Patients

    Journal: Frontiers in Neuroanatomy

    doi: 10.3389/fnana.2010.00020

    Correlative light and electron microscopy of PHF-tau-ir neurons in the pyramidal layer of CA1 from patient P1 . (A) Photomicrograph of a plastic-embedded 100 μm-thick section immunostained for PHF-tau. (B) Photomicrograph of a 2-μm-thick semithin section stained with 1% toluidine blue obtained from the section shown in (A) . (C) , (F) , higher magnification of cells 1 (pattern I of PHF-tau-immunostaining) and 7 (pattern II of PHF-tau-immunostaining), respectively, also shown in (A) and (B) . (D) , (E,G) , electron micrographs of neurons 1 and 7, respectively, obtained after the resectioning of the semithin section shown in (B) . The cytoplasm of neuron 1 (D) has a normal appearance, whereas the cytoplasm of neuron 7 (G) and (E) has NFT of paired helical filaments stained for PHF-tau (asterisks in E and I ). (H,I,J) Electron micrographs illustrating the neuropil around PHF-tau-ir cells. Arrows indicate some normal looking synaptic contacts situated in close proximity of the PHF-tau-ir cell bodies which have been pseudo-colored in blue. Scale bar: (A) , 104 μm; (B) , 95 μm; (C , F) , 23 μm; (D) , 1.5 μm; (E) , 1.1 μm; (G) , 10 μm; (H) , 0.6 μm; (I) , 0.8 μm; (J) , 0.5 μm.
    Figure Legend Snippet: Correlative light and electron microscopy of PHF-tau-ir neurons in the pyramidal layer of CA1 from patient P1 . (A) Photomicrograph of a plastic-embedded 100 μm-thick section immunostained for PHF-tau. (B) Photomicrograph of a 2-μm-thick semithin section stained with 1% toluidine blue obtained from the section shown in (A) . (C) , (F) , higher magnification of cells 1 (pattern I of PHF-tau-immunostaining) and 7 (pattern II of PHF-tau-immunostaining), respectively, also shown in (A) and (B) . (D) , (E,G) , electron micrographs of neurons 1 and 7, respectively, obtained after the resectioning of the semithin section shown in (B) . The cytoplasm of neuron 1 (D) has a normal appearance, whereas the cytoplasm of neuron 7 (G) and (E) has NFT of paired helical filaments stained for PHF-tau (asterisks in E and I ). (H,I,J) Electron micrographs illustrating the neuropil around PHF-tau-ir cells. Arrows indicate some normal looking synaptic contacts situated in close proximity of the PHF-tau-ir cell bodies which have been pseudo-colored in blue. Scale bar: (A) , 104 μm; (B) , 95 μm; (C , F) , 23 μm; (D) , 1.5 μm; (E) , 1.1 μm; (G) , 10 μm; (H) , 0.6 μm; (I) , 0.8 μm; (J) , 0.5 μm.

    Techniques Used: Electron Microscopy, Staining, Immunostaining

    2) Product Images from "Arsenic Exposure Increases Monocyte Adhesion to the Vascular Endothelium, a Pro-Atherogenic Mechanism"

    Article Title: Arsenic Exposure Increases Monocyte Adhesion to the Vascular Endothelium, a Pro-Atherogenic Mechanism

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0136592

    Arsenic induces monocyte adhesion to endothelial cells, with maximal binding achieved following exposure of both cell types. (A) U937 and/or HUVEC cells (1000 cells/ml) were exposed to arsenic overnight (0, 10 or 200 ppb). U937 cells were fluorescently-labelled and were incubated with HUVEC cells. The non-adhered cells were washed away, and the adherent fluorescent cells were counted. Data are expressed as relative number of U937 over total HUVEC stained cells. ** = p
    Figure Legend Snippet: Arsenic induces monocyte adhesion to endothelial cells, with maximal binding achieved following exposure of both cell types. (A) U937 and/or HUVEC cells (1000 cells/ml) were exposed to arsenic overnight (0, 10 or 200 ppb). U937 cells were fluorescently-labelled and were incubated with HUVEC cells. The non-adhered cells were washed away, and the adherent fluorescent cells were counted. Data are expressed as relative number of U937 over total HUVEC stained cells. ** = p

    Techniques Used: Binding Assay, Incubation, Staining

    3) Product Images from "Early maturation and distinct tau pathology in induced pluripotent stem cell-derived neurons from patients with MAPT mutations"

    Article Title: Early maturation and distinct tau pathology in induced pluripotent stem cell-derived neurons from patients with MAPT mutations

    Journal: Brain

    doi: 10.1093/brain/awv222

    Immunohistochemistry of brain sections from the cortex of the patient with the P301L mutation and a control subject. ( A ) Confocal microscope image of control and P301L human brain frontal cortex sections stained with anti-βIII-tubulin antibody. The arrow points to a varicosity-like structure. No similar structures were found in the human control brain. ( B ) Examples of varicosity-like structures stained by phosphorylation-dependent anti-tau antibody AT8, in the frontal cortex of the P301L patient from whom IPSCs were generated. Red arrows point to varicosity-like structures. Blue arrows point to cell bodies where nuclei are stained by cresyl violet. Scale bars: A = 17 μm; B = 68 µm.
    Figure Legend Snippet: Immunohistochemistry of brain sections from the cortex of the patient with the P301L mutation and a control subject. ( A ) Confocal microscope image of control and P301L human brain frontal cortex sections stained with anti-βIII-tubulin antibody. The arrow points to a varicosity-like structure. No similar structures were found in the human control brain. ( B ) Examples of varicosity-like structures stained by phosphorylation-dependent anti-tau antibody AT8, in the frontal cortex of the P301L patient from whom IPSCs were generated. Red arrows point to varicosity-like structures. Blue arrows point to cell bodies where nuclei are stained by cresyl violet. Scale bars: A = 17 μm; B = 68 µm.

    Techniques Used: Immunohistochemistry, Mutagenesis, Microscopy, Staining, Generated

    Tau hyperphosphorylation and morphological characterization of IPSC-derived neurons. ( A ) Immunocytochemistry of control, P301L and N279K IPSC-derived neurons at Day 55 using the phosphorylation-dependent anti-tau antibody AT8 (green), βIII-tubulin (red) and Hoechst (blue). Staining for AT8 is present in several P301L and N279K neurons where it often consists of ring-like structures. ( B ) Immunocytochemistry for phosphorylation-dependent anti-tau antibody AT100 (green), βIII-tubulin (red) and Hoechst (blue) in control, P301L and N279K neurons at Day 150. Some N279K neurons show AT100 staining in cell bodies. The image magnification in the insert shows accumulation of tau protein stained by AT100 in neurites and in cell bodies where sometimes the staining has a dotted appearance. ( C ) Statistical analysis of the AT8-positive neurons within the βIII-tubulin positive cells shows a significant increase in tau hyperphosphorylation in P301L and N279K neurons compared to controls (CRL). Cell count was performed using ImageJ and statistical analysis was evaluated with Student’s t -test. ** P
    Figure Legend Snippet: Tau hyperphosphorylation and morphological characterization of IPSC-derived neurons. ( A ) Immunocytochemistry of control, P301L and N279K IPSC-derived neurons at Day 55 using the phosphorylation-dependent anti-tau antibody AT8 (green), βIII-tubulin (red) and Hoechst (blue). Staining for AT8 is present in several P301L and N279K neurons where it often consists of ring-like structures. ( B ) Immunocytochemistry for phosphorylation-dependent anti-tau antibody AT100 (green), βIII-tubulin (red) and Hoechst (blue) in control, P301L and N279K neurons at Day 150. Some N279K neurons show AT100 staining in cell bodies. The image magnification in the insert shows accumulation of tau protein stained by AT100 in neurites and in cell bodies where sometimes the staining has a dotted appearance. ( C ) Statistical analysis of the AT8-positive neurons within the βIII-tubulin positive cells shows a significant increase in tau hyperphosphorylation in P301L and N279K neurons compared to controls (CRL). Cell count was performed using ImageJ and statistical analysis was evaluated with Student’s t -test. ** P

    Techniques Used: Derivative Assay, Immunocytochemistry, Staining, Cell Counting

    4) Product Images from "Preclinical ex-vivo Testing of Anti-inflammatory Drugs in a Bovine Intervertebral Degenerative Disc Model"

    Article Title: Preclinical ex-vivo Testing of Anti-inflammatory Drugs in a Bovine Intervertebral Degenerative Disc Model

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2020.00583

    IL-1β immunohistochemistry staining of IVD tissue from the Etanercept experiments. Representative IL-1β IHC image of NP region, inner AF region (iAF) and outer AF (oAF) region from day 0 control samples (Day0.inj.), IVDs cultured under physiological condition on day 4 (Phy.inj.), IVDs cultured under degenerative condition on day 4 (Deg.inj.), and IVDs cultured under degenerative condition and treated with Etanercept on day 4 (Etanercept.inj.). Scale bar: 100 μm. The percentage of positively stained cells were counted, as presented in the bar graph. n = 8, Means + SEM, * p
    Figure Legend Snippet: IL-1β immunohistochemistry staining of IVD tissue from the Etanercept experiments. Representative IL-1β IHC image of NP region, inner AF region (iAF) and outer AF (oAF) region from day 0 control samples (Day0.inj.), IVDs cultured under physiological condition on day 4 (Phy.inj.), IVDs cultured under degenerative condition on day 4 (Deg.inj.), and IVDs cultured under degenerative condition and treated with Etanercept on day 4 (Etanercept.inj.). Scale bar: 100 μm. The percentage of positively stained cells were counted, as presented in the bar graph. n = 8, Means + SEM, * p

    Techniques Used: Immunohistochemistry, Staining, Cell Culture

    IL-1β immunohistochemistry staining of IVD tissue from the Tofacitinib experiments. Representative IL-1β IHC image of NP region, inner AF region (iAF) and outer AF (oAF) region from day 0 control samples (Day0.med.), IVDs cultured under physiological condition on day 4 (Phy.med.), IVDs cultured under degenerative condition on day 4 (Deg.med.), and IVDs cultured under degenerative condition and treated with Tofacitinib on day 4 (Tofacitinib.med.). Scale bar: 100 μm. The percentage of positively stained cells were counted, as presented in the bar graph. n = 8, Means + SEM, ** p
    Figure Legend Snippet: IL-1β immunohistochemistry staining of IVD tissue from the Tofacitinib experiments. Representative IL-1β IHC image of NP region, inner AF region (iAF) and outer AF (oAF) region from day 0 control samples (Day0.med.), IVDs cultured under physiological condition on day 4 (Phy.med.), IVDs cultured under degenerative condition on day 4 (Deg.med.), and IVDs cultured under degenerative condition and treated with Tofacitinib on day 4 (Tofacitinib.med.). Scale bar: 100 μm. The percentage of positively stained cells were counted, as presented in the bar graph. n = 8, Means + SEM, ** p

    Techniques Used: Immunohistochemistry, Staining, Cell Culture

    5) Product Images from "Cdk8 is required for establishment of H3K27me3 and gene repression by Xist and mouse development"

    Article Title: Cdk8 is required for establishment of H3K27me3 and gene repression by Xist and mouse development

    Journal: Development (Cambridge, England)

    doi: 10.1242/dev.175141

    Cdk8 is required for efficient PRC2 recruitment by Xist . (A) Images of combined H2AK119ub immunofluorescence with Xist RNA FISH (red) for wild-type and ΔCdk8 cells. DAPI was used to stain DNA (blue). (B) Quantification of the percentage of Xist clusters with H2AK119ub foci after 24 h of Xist expression in ΔCdk8 and wild-type ESCs. Percentages are relative to counted Xist clusters; experiments were performed in triplicate; data are mean±s.d. (C) Combined immunofluorescence (Ezh2 and H3K27me3) with Xist -FISH (red) for wild-type and ΔCdk8 ESCs. White arrows indicate Xist clusters lacking PRC2 marks. DNA was stained with DAPI (blue). (D-F) Quantification of the percentage of Xist clusters with PRC2 (Ezh2 and H3K27me3) foci after 24 h of Xist expression in (D) ΔCdk8 ESCs and wild-type Cdk8 transgene complemented ΔCdk8 ESCs, (E) D151A and (F) ΔATP mutant Cdk8 transgene complemented ΔCdk8 ESCs. Percentages are relative to counted Xist clusters. Wild-type, ΔCdk8 #8 and ΔCdk8 #8 TG#15 samples are the same in E and F. The experiments were performed in triplicate. Data are mean±s.d.; asterisk indicates significant changes relative to WT ( P
    Figure Legend Snippet: Cdk8 is required for efficient PRC2 recruitment by Xist . (A) Images of combined H2AK119ub immunofluorescence with Xist RNA FISH (red) for wild-type and ΔCdk8 cells. DAPI was used to stain DNA (blue). (B) Quantification of the percentage of Xist clusters with H2AK119ub foci after 24 h of Xist expression in ΔCdk8 and wild-type ESCs. Percentages are relative to counted Xist clusters; experiments were performed in triplicate; data are mean±s.d. (C) Combined immunofluorescence (Ezh2 and H3K27me3) with Xist -FISH (red) for wild-type and ΔCdk8 ESCs. White arrows indicate Xist clusters lacking PRC2 marks. DNA was stained with DAPI (blue). (D-F) Quantification of the percentage of Xist clusters with PRC2 (Ezh2 and H3K27me3) foci after 24 h of Xist expression in (D) ΔCdk8 ESCs and wild-type Cdk8 transgene complemented ΔCdk8 ESCs, (E) D151A and (F) ΔATP mutant Cdk8 transgene complemented ΔCdk8 ESCs. Percentages are relative to counted Xist clusters. Wild-type, ΔCdk8 #8 and ΔCdk8 #8 TG#15 samples are the same in E and F. The experiments were performed in triplicate. Data are mean±s.d.; asterisk indicates significant changes relative to WT ( P

    Techniques Used: Immunofluorescence, Fluorescence In Situ Hybridization, Staining, Expressing, Mutagenesis

    Cdk8 contributes to the initiation of XCI in female mouse development. (A) Western blot analysis of Cdk8 with β-Actin as loading control in ESCs established from embryos of crosses between Cdk8 WT/1lox Sox2-Cre +/− males and Cdk8 2lox/2lox females. (B) RNA FISH expression analysis of Xist and X-linked genes Lamp2 (middle row) or G6pdx (bottom row) in homozygous female Cdk8 1lox and control female ESCs after 96 h RA differentiation. Top row, Xist (red); middle and bottom rows, Xist (green) and X-linked genes (red), Lamp2 (middle) and G6pdx (bottom) RNA FISH. DNA was stained with DAPI (blue). (C) Quantification of Xist RNA FISH analysis of Cdk8 1lox and control (WT/2LOX) female ESCs after 96 h RA differentiation. The percentage of Xist clusters is shown relative to counted nuclei ( > 100 counted). The experiments were performed in triplicate. Data are mean±s.d. a.u., arbitrary units. (D) Quantification of the fluorescence intensity of Xist clusters from RNA FISH ( > 100 clusters measured). The experiments were performed in triplicate. Data are mean±s.d. (E) Double FISH quantification of biallelic expression of the X-linked genes Lamp2 and G6pdx , and overlap with an Xist cluster. Percentage ( > 100 Xist clusters analysed) of Xist clusters without detectable biallelic X-linked gene expression (green bars), and Xist clusters with overlapping X-linked gene expression (red bars) are shown. The experiment was performed in triplicate. Data are mean±s.d. Asterisk indicates a significant difference from the wild-type control ( P
    Figure Legend Snippet: Cdk8 contributes to the initiation of XCI in female mouse development. (A) Western blot analysis of Cdk8 with β-Actin as loading control in ESCs established from embryos of crosses between Cdk8 WT/1lox Sox2-Cre +/− males and Cdk8 2lox/2lox females. (B) RNA FISH expression analysis of Xist and X-linked genes Lamp2 (middle row) or G6pdx (bottom row) in homozygous female Cdk8 1lox and control female ESCs after 96 h RA differentiation. Top row, Xist (red); middle and bottom rows, Xist (green) and X-linked genes (red), Lamp2 (middle) and G6pdx (bottom) RNA FISH. DNA was stained with DAPI (blue). (C) Quantification of Xist RNA FISH analysis of Cdk8 1lox and control (WT/2LOX) female ESCs after 96 h RA differentiation. The percentage of Xist clusters is shown relative to counted nuclei ( > 100 counted). The experiments were performed in triplicate. Data are mean±s.d. a.u., arbitrary units. (D) Quantification of the fluorescence intensity of Xist clusters from RNA FISH ( > 100 clusters measured). The experiments were performed in triplicate. Data are mean±s.d. (E) Double FISH quantification of biallelic expression of the X-linked genes Lamp2 and G6pdx , and overlap with an Xist cluster. Percentage ( > 100 Xist clusters analysed) of Xist clusters without detectable biallelic X-linked gene expression (green bars), and Xist clusters with overlapping X-linked gene expression (red bars) are shown. The experiment was performed in triplicate. Data are mean±s.d. Asterisk indicates a significant difference from the wild-type control ( P

    Techniques Used: Western Blot, Fluorescence In Situ Hybridization, Expressing, Staining, Fluorescence

    6) Product Images from "Identification of oligomers at early stages of tau aggregation in Alzheimer's disease"

    Article Title: Identification of oligomers at early stages of tau aggregation in Alzheimer's disease

    Journal: The FASEB Journal

    doi: 10.1096/fj.11-199851

    Ubiquitination of tau occurs at intermediate stages during NFT evolution, after appearance of oligomers. A–C ) Immunofluorescence images of pretangles fail to show colocalization of anti-ubiquitin staining ( A , green), T22 ( B , red), and merge (
    Figure Legend Snippet: Ubiquitination of tau occurs at intermediate stages during NFT evolution, after appearance of oligomers. A–C ) Immunofluorescence images of pretangles fail to show colocalization of anti-ubiquitin staining ( A , green), T22 ( B , red), and merge (

    Techniques Used: Immunofluorescence, Staining

    7) Product Images from "Autologous skeletal muscle derived cells expressing a novel functional dystrophin provide a potential therapy for Duchenne Muscular Dystrophy"

    Article Title: Autologous skeletal muscle derived cells expressing a novel functional dystrophin provide a potential therapy for Duchenne Muscular Dystrophy

    Journal: Scientific Reports

    doi: 10.1038/srep19750

    Restoration of dystrophin in regenerated muscle fibres derived from lentivirally-transduced DMD pericytes. ( A ) shows representative transverse sections of cryodamaged muscles of an mdx nude mouse that had been grafted with a–d: SFFV-C1-GFP cells; e–h: hDesmin-C1-GFP cells. Muscles transplanted with SFFV-C2-GFP and hDesmin-C2-GFP cells showed similar staining pattern as that shown in a–d. Sections were stained with antibodies to human spectrin (red), GFP (green), dystrophin (cyan). Nuclei were counterstained with DAPI (blue). Scale bar = 15 μm. ( B ) and ( C ) are the quantification of human Spectrin+ fibres ( B ) and GFP+ fibres ( C ) in SFFV-C1-GFP, hDesmin-C1-GFP, SFFV-C2-GFP and hDesmin-C2-GFP cells transplanted groups. * p
    Figure Legend Snippet: Restoration of dystrophin in regenerated muscle fibres derived from lentivirally-transduced DMD pericytes. ( A ) shows representative transverse sections of cryodamaged muscles of an mdx nude mouse that had been grafted with a–d: SFFV-C1-GFP cells; e–h: hDesmin-C1-GFP cells. Muscles transplanted with SFFV-C2-GFP and hDesmin-C2-GFP cells showed similar staining pattern as that shown in a–d. Sections were stained with antibodies to human spectrin (red), GFP (green), dystrophin (cyan). Nuclei were counterstained with DAPI (blue). Scale bar = 15 μm. ( B ) and ( C ) are the quantification of human Spectrin+ fibres ( B ) and GFP+ fibres ( C ) in SFFV-C1-GFP, hDesmin-C1-GFP, SFFV-C2-GFP and hDesmin-C2-GFP cells transplanted groups. * p

    Techniques Used: Derivative Assay, Staining

    8) Product Images from "High Throughput Screening for Compounds That Alter Muscle Cell Glycosylation Identifies New Role for N-Glycans in Regulating Sarcolemmal Protein Abundance and Laminin Binding *"

    Article Title: High Throughput Screening for Compounds That Alter Muscle Cell Glycosylation Identifies New Role for N-Glycans in Regulating Sarcolemmal Protein Abundance and Laminin Binding *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.334581

    Lobeline increases abundance of multiple muscle cell proteins. A , wild type ( wt ) primary myoblasts differentiated in the presence of 100 μ m lobeline ( L ) or control ( C ) were solubilized in radioimmune precipitation assay buffer, and 60 μg of protein was separated by SDS-PAGE and blotted to nitrocellulose. Membranes were probed with antibodies against agrin, dystrophin, utrophin, dystroglycans (α- and β-DG), sarcospan ( SSPN ), sarcoglycans (α-, β-, and γ-SG), β1D integrin, dysferlin, and caveolin-3. GAPDH was a loading control. Lobeline treatment increased agrin, integrin, dysferlin, DGC, and UGC protein levels. B , mdx primary myoblasts were treated as described in A . Membranes were probed similarly with the addition of antibody to β-SG. Dystrophin protein was not detected due to mutation in the dystrophin gene in mdx cells. Increases were observed for agrin, dysferlin, utrophin, and α- and β-dystroglycans.
    Figure Legend Snippet: Lobeline increases abundance of multiple muscle cell proteins. A , wild type ( wt ) primary myoblasts differentiated in the presence of 100 μ m lobeline ( L ) or control ( C ) were solubilized in radioimmune precipitation assay buffer, and 60 μg of protein was separated by SDS-PAGE and blotted to nitrocellulose. Membranes were probed with antibodies against agrin, dystrophin, utrophin, dystroglycans (α- and β-DG), sarcospan ( SSPN ), sarcoglycans (α-, β-, and γ-SG), β1D integrin, dysferlin, and caveolin-3. GAPDH was a loading control. Lobeline treatment increased agrin, integrin, dysferlin, DGC, and UGC protein levels. B , mdx primary myoblasts were treated as described in A . Membranes were probed similarly with the addition of antibody to β-SG. Dystrophin protein was not detected due to mutation in the dystrophin gene in mdx cells. Increases were observed for agrin, dysferlin, utrophin, and α- and β-dystroglycans.

    Techniques Used: SDS Page, Mutagenesis

    9) Product Images from "Homodimerization Is Essential for the Receptor for Advanced Glycation End Products (RAGE)-mediated Signal Transduction *"

    Article Title: Homodimerization Is Essential for the Receptor for Advanced Glycation End Products (RAGE)-mediated Signal Transduction *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.133827

    RAGE-activated MAPK pathways in HEK293T cells. A , 4% paraformaldehyde-fixed HEK293T cells were evaluated for RAGE immunolocalization using an anti-RAGE monoclonal antibody ( Ab ) ( green ). The cell nuclei were stained with 4′,6-diamidino-2-phenylindole
    Figure Legend Snippet: RAGE-activated MAPK pathways in HEK293T cells. A , 4% paraformaldehyde-fixed HEK293T cells were evaluated for RAGE immunolocalization using an anti-RAGE monoclonal antibody ( Ab ) ( green ). The cell nuclei were stained with 4′,6-diamidino-2-phenylindole

    Techniques Used: Staining

    10) Product Images from "Nitric oxide selectively suppresses IH currents mediated by HCN1-containing channels"

    Article Title: Nitric oxide selectively suppresses IH currents mediated by HCN1-containing channels

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2014.282194

    Expression of HCN1 and HCN2 within the mouse SOC A , intense immunofluorescence for HCN1 (green) was predominant in cell bodies and fibres in the MSO and LSO, sparse in the SPN and absent in the MNTB, where immunofluorescence was of background level. Micrographs taken at high magnification show immunopositive cell soma in the LSO ( A i ), MSO ( A ii ) and SPN ( A iii ) but MNTB principal neurons showed little or no specific labelling ( A iv ; HCN1 = green; different sections to A ). B , HCN2 immunofluorescence (green) was most intense in cell bodies and fibres in the MNTB and SPN, weaker in the LSO and comparable to background in the MSO. B i–iv , high magnification images show the presence of immunopositive cell soma in the LSO ( B i ), SPN ( B iii ) and MNTB ( B iv ) but not MSO ( B ii ; green = HCN2; different sections to B ). Asterisks (*) in B iv mark principal cell bodies. C , control sections for the specificity of the secondary antibody (primary antibodies omitted) showed little non-specific staining (green). Dashed line surrounds MNTB. A , B and C are composites of two photographs. Tissue sections shown in A–A iv were adjacent to those shown in B–B iv . Cell nuclei were counterstained with DAPI (blue). Scale bar in A = 150 μm for A , B and C and 25 μm for A i–iv and B i–iv .
    Figure Legend Snippet: Expression of HCN1 and HCN2 within the mouse SOC A , intense immunofluorescence for HCN1 (green) was predominant in cell bodies and fibres in the MSO and LSO, sparse in the SPN and absent in the MNTB, where immunofluorescence was of background level. Micrographs taken at high magnification show immunopositive cell soma in the LSO ( A i ), MSO ( A ii ) and SPN ( A iii ) but MNTB principal neurons showed little or no specific labelling ( A iv ; HCN1 = green; different sections to A ). B , HCN2 immunofluorescence (green) was most intense in cell bodies and fibres in the MNTB and SPN, weaker in the LSO and comparable to background in the MSO. B i–iv , high magnification images show the presence of immunopositive cell soma in the LSO ( B i ), SPN ( B iii ) and MNTB ( B iv ) but not MSO ( B ii ; green = HCN2; different sections to B ). Asterisks (*) in B iv mark principal cell bodies. C , control sections for the specificity of the secondary antibody (primary antibodies omitted) showed little non-specific staining (green). Dashed line surrounds MNTB. A , B and C are composites of two photographs. Tissue sections shown in A–A iv were adjacent to those shown in B–B iv . Cell nuclei were counterstained with DAPI (blue). Scale bar in A = 150 μm for A , B and C and 25 μm for A i–iv and B i–iv .

    Techniques Used: Expressing, Immunofluorescence, Staining

    nNOS immunostaining predominates in the MNTB and SPN A , immunofluorescence for nNOS (green) in tissue from a wild-type (WT) mouse was apparent in cell soma, fibres (arrows) and neuropil throughout the MNTB, SPN, MSO and LSO. B , identical staining procedures applied to tissue from a nNOS knockout (KO) mouse showed little or no staining (green), thereby validating the specificity of the immunofluorescence for nNOS in the WT. Inset: secondary antibody control (no primary antibody was used). Image shows MNTB in a knockout section and is representative of the entire SOC. A and B are merged composites of two images to show the full lateral extent of the SOC. Nuclei were counterstained with DAPI (blue). Scale bar = 150 μm and applies to both panels and inset.
    Figure Legend Snippet: nNOS immunostaining predominates in the MNTB and SPN A , immunofluorescence for nNOS (green) in tissue from a wild-type (WT) mouse was apparent in cell soma, fibres (arrows) and neuropil throughout the MNTB, SPN, MSO and LSO. B , identical staining procedures applied to tissue from a nNOS knockout (KO) mouse showed little or no staining (green), thereby validating the specificity of the immunofluorescence for nNOS in the WT. Inset: secondary antibody control (no primary antibody was used). Image shows MNTB in a knockout section and is representative of the entire SOC. A and B are merged composites of two images to show the full lateral extent of the SOC. Nuclei were counterstained with DAPI (blue). Scale bar = 150 μm and applies to both panels and inset.

    Techniques Used: Immunostaining, Immunofluorescence, Staining, Knock-Out

    11) Product Images from "Therapeutic Efficacy of p53 Restoration in Mdm2-overexpressing Tumors"

    Article Title: Therapeutic Efficacy of p53 Restoration in Mdm2-overexpressing Tumors

    Journal: Molecular cancer research : MCR

    doi: 10.1158/1541-7786.MCR-14-0089

    Reduced cell proliferation and induced cellular senescence in Mdm2 Tg p53 Neo/Neo CreER angiosarcomas upon p53 restoration Tamoxifen-treated angiosarcomas collected from the MRI study were subjected to immunohistochemical staining for proliferation marker Ki-67, cellular senescence marker PML, and apoptotic marker cleaved caspase-3 as well as H E staining. a, d, g and J) Image analysis of the representative Mdm2 Tg p53 Neo/Neo angiosarcomas treated with multiple doses of tamoxifen; b, e, h and k) Image analysis of the representative Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with one dose of tamoxifen; c, f, i and l) Image analysis of the representative Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with four doses of tamoxifen. m) Percentages of cells staining positive for Ki-67 in tamoxifen-treated angiosarcomas. Four random fields (40 magnification) for each tumor were chosen to count the positive cells.
    Figure Legend Snippet: Reduced cell proliferation and induced cellular senescence in Mdm2 Tg p53 Neo/Neo CreER angiosarcomas upon p53 restoration Tamoxifen-treated angiosarcomas collected from the MRI study were subjected to immunohistochemical staining for proliferation marker Ki-67, cellular senescence marker PML, and apoptotic marker cleaved caspase-3 as well as H E staining. a, d, g and J) Image analysis of the representative Mdm2 Tg p53 Neo/Neo angiosarcomas treated with multiple doses of tamoxifen; b, e, h and k) Image analysis of the representative Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with one dose of tamoxifen; c, f, i and l) Image analysis of the representative Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with four doses of tamoxifen. m) Percentages of cells staining positive for Ki-67 in tamoxifen-treated angiosarcomas. Four random fields (40 magnification) for each tumor were chosen to count the positive cells.

    Techniques Used: Magnetic Resonance Imaging, Immunohistochemistry, Staining, Marker

    Restoration of p53 expression in Mdm2 Tg p53 Neo/Neo CreER MEFs a) qRT-PCR analysis of p53 mRNA levels in the Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1µM 4-hydroxytamoxifen or vehicle control. Data were normalized to expression in vehicle-treated control and represent a mean± SEM (n=3). b) MTT assays of Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1µM 4-hydroxytamoxifen or vehicle control. Data were normalized to absorbance values at day 1 of each treatment and represent a mean of three independent experiments ± SEM. c) qRT-PCR analysis of expression of p53 target genes p21 and Noxa in the Mdm2 Tg p53 Neo/Neo CreER MEFs. Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1 µM 4-hydroxytamoxifen or vehicle control for 48 hours, followed by doxorubicin (1 µg/mL) treatment for 4 hours or vehicle treatment. Data were normalized to expression in vehicle-treated control and represent a mean± SEM (n=3). d) Western blot analysis of p53 expression and e) qRT-PCR analysis of expression of p53 target genes p21 and Noxa in H-Ras G12V -infected Mdm2 Tg p53 Neo/Neo CreER MEFs upon 4-hydroxytamoxifen treatment for 48 hours. Data were normalized to expression in vehicle-treated control cells without H-Ras G12V expression and represent a mean± SEM (n=3). f) MTT assays of H-Ras G12V -infected Mdm2 Tg p53 Neo/Neo CreER MEFs incubated with 1 µM 4-hydroxytamoxifen or vehicle control. Data were normalized to absorbance values at day 1 of each treatment and represent a mean of three independent experiments ± SEM. 4OH-Tam, 4-hydroxytamoxifen; Doxo, doxorubicin; H-Ras, H-Ras G12V .
    Figure Legend Snippet: Restoration of p53 expression in Mdm2 Tg p53 Neo/Neo CreER MEFs a) qRT-PCR analysis of p53 mRNA levels in the Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1µM 4-hydroxytamoxifen or vehicle control. Data were normalized to expression in vehicle-treated control and represent a mean± SEM (n=3). b) MTT assays of Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1µM 4-hydroxytamoxifen or vehicle control. Data were normalized to absorbance values at day 1 of each treatment and represent a mean of three independent experiments ± SEM. c) qRT-PCR analysis of expression of p53 target genes p21 and Noxa in the Mdm2 Tg p53 Neo/Neo CreER MEFs. Mdm2 Tg p53 Neo/Neo CreER MEFs treated with 1 µM 4-hydroxytamoxifen or vehicle control for 48 hours, followed by doxorubicin (1 µg/mL) treatment for 4 hours or vehicle treatment. Data were normalized to expression in vehicle-treated control and represent a mean± SEM (n=3). d) Western blot analysis of p53 expression and e) qRT-PCR analysis of expression of p53 target genes p21 and Noxa in H-Ras G12V -infected Mdm2 Tg p53 Neo/Neo CreER MEFs upon 4-hydroxytamoxifen treatment for 48 hours. Data were normalized to expression in vehicle-treated control cells without H-Ras G12V expression and represent a mean± SEM (n=3). f) MTT assays of H-Ras G12V -infected Mdm2 Tg p53 Neo/Neo CreER MEFs incubated with 1 µM 4-hydroxytamoxifen or vehicle control. Data were normalized to absorbance values at day 1 of each treatment and represent a mean of three independent experiments ± SEM. 4OH-Tam, 4-hydroxytamoxifen; Doxo, doxorubicin; H-Ras, H-Ras G12V .

    Techniques Used: Expressing, Quantitative RT-PCR, MTT Assay, Western Blot, Infection, Incubation

    Survival and tumor volume changes in angiosarcomas of Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice with tamoxifen treatment a) Survival curves of Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice with angiosarcomas upon tamoxifen treatment. b) Tumor volume changes (post-treatment volume/pre-treatment volume ratios) of angiosarcomas in Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice during tamoxifen treatment. c) MRI images and changes in tumor volumes of two representative angiosarcomas (1 Mdm2 Tg p53 Neo/Neo CreER and 1 Mdm2 Tg p53 Neo/Neo ) during tamoxifen treatment. Day 0, the day when tamoxifen treatment started; Day 28, 28 days after initial tamoxifen treatment, etc .
    Figure Legend Snippet: Survival and tumor volume changes in angiosarcomas of Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice with tamoxifen treatment a) Survival curves of Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice with angiosarcomas upon tamoxifen treatment. b) Tumor volume changes (post-treatment volume/pre-treatment volume ratios) of angiosarcomas in Mdm2 Tg p53 Neo/Neo CreER and Mdm2 Tg p53 Neo/Neo mice during tamoxifen treatment. c) MRI images and changes in tumor volumes of two representative angiosarcomas (1 Mdm2 Tg p53 Neo/Neo CreER and 1 Mdm2 Tg p53 Neo/Neo ) during tamoxifen treatment. Day 0, the day when tamoxifen treatment started; Day 28, 28 days after initial tamoxifen treatment, etc .

    Techniques Used: Mouse Assay, Magnetic Resonance Imaging

    p53 restoration in a syngeneic transplant model of Mdm2 Tg p53 Neo/Neo CreER angiosarcoma a) Growth curves of transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen (n=8) and vehicle control (n=3). b) Comparison of tumor volume doubling time calculated from tumor growth curves between tamoxifen-treated and vehicle control groups. Tumor volume doubling time was calculated based on the exponential curve fit of tumor volume against time. c) Western blot analysis of p53 and p21 levels in transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen and vehicle control. Three representative tumors from both control and tamoxifen-treated groups were examined. Protein lysate from an Mdm2 −/− p53 −/− spleen served as negative control. d) qRT-PCR analysis of expression of p53 target genes p21 and Puma in transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen and vehicle control. Data were normalized to expression in a vehicle-treated control.
    Figure Legend Snippet: p53 restoration in a syngeneic transplant model of Mdm2 Tg p53 Neo/Neo CreER angiosarcoma a) Growth curves of transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen (n=8) and vehicle control (n=3). b) Comparison of tumor volume doubling time calculated from tumor growth curves between tamoxifen-treated and vehicle control groups. Tumor volume doubling time was calculated based on the exponential curve fit of tumor volume against time. c) Western blot analysis of p53 and p21 levels in transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen and vehicle control. Three representative tumors from both control and tamoxifen-treated groups were examined. Protein lysate from an Mdm2 −/− p53 −/− spleen served as negative control. d) qRT-PCR analysis of expression of p53 target genes p21 and Puma in transplanted Mdm2 Tg p53 Neo/Neo CreER angiosarcomas treated with tamoxifen and vehicle control. Data were normalized to expression in a vehicle-treated control.

    Techniques Used: Western Blot, Negative Control, Quantitative RT-PCR, Expressing

    12) Product Images from "Selective Estrogen Receptor Modulators Enhance CNS Remyelination Independent of Estrogen Receptors"

    Article Title: Selective Estrogen Receptor Modulators Enhance CNS Remyelination Independent of Estrogen Receptors

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1530-18.2019

    BZA enhances and accelerates remyelination in an in vivo focal demyelination model a , Schematic showing location of lysolecithin injection into the corpus callosum at the position: AP: −1.04, ML: 1, and DV: −1.75 from bregma. b , Representative images of lesions within the corpus callosum of adult WT mice. Lesions were identified by the density of cell nuclei (DAPI) and included based on NF staining for intact axons and their boundaries are demarcated in white. Animals were given either vehicle or BZA at 10 mg/kg/d for 7 d beginning at 1 dpl and perfused at 10 dpl. They were immunostained for CC1 (red) and MOG (green). Cell nuclei are shown by DAPI (blue). c , Quantification of CC1 + cell density. d , Quantification of MOG as a percentage of area. e , Representative images of myelinated axon density in nonlesioned areas in vehicle- and BZA-treated mice. f , Scatterplot of axon diameter plotted against corresponding g-ratio of vehicle- and BZA-treated mice demonstrating no difference in g-ratio between groups. g , Quantification of myelinated axon density within the nonlesioned area. h , Quantification of average g-ratio within lesioned and nonlesioned area. i , Representative images of remyelinated axon density within the lesioned area in vehicle- and BZA-treated mice. j , Scatterplot of axon diameter plotted against corresponding g-ratio of vehicle- and BZA-treated mice in the lesioned area. k , Quantification of remyelinated axon density within lesioned areas in vehicle- and BZA-treated mice. l , Quantification of the percentage of unmyelinated axons in lesion areas in vehicle- and BZA-treated mice. Error bars indicate SEM. ** p
    Figure Legend Snippet: BZA enhances and accelerates remyelination in an in vivo focal demyelination model a , Schematic showing location of lysolecithin injection into the corpus callosum at the position: AP: −1.04, ML: 1, and DV: −1.75 from bregma. b , Representative images of lesions within the corpus callosum of adult WT mice. Lesions were identified by the density of cell nuclei (DAPI) and included based on NF staining for intact axons and their boundaries are demarcated in white. Animals were given either vehicle or BZA at 10 mg/kg/d for 7 d beginning at 1 dpl and perfused at 10 dpl. They were immunostained for CC1 (red) and MOG (green). Cell nuclei are shown by DAPI (blue). c , Quantification of CC1 + cell density. d , Quantification of MOG as a percentage of area. e , Representative images of myelinated axon density in nonlesioned areas in vehicle- and BZA-treated mice. f , Scatterplot of axon diameter plotted against corresponding g-ratio of vehicle- and BZA-treated mice demonstrating no difference in g-ratio between groups. g , Quantification of myelinated axon density within the nonlesioned area. h , Quantification of average g-ratio within lesioned and nonlesioned area. i , Representative images of remyelinated axon density within the lesioned area in vehicle- and BZA-treated mice. j , Scatterplot of axon diameter plotted against corresponding g-ratio of vehicle- and BZA-treated mice in the lesioned area. k , Quantification of remyelinated axon density within lesioned areas in vehicle- and BZA-treated mice. l , Quantification of the percentage of unmyelinated axons in lesion areas in vehicle- and BZA-treated mice. Error bars indicate SEM. ** p

    Techniques Used: In Vivo, Injection, Mouse Assay, Staining

    13) Product Images from "Effect of high WDR5 expression on the hepatocellular carcinoma prognosis"

    Article Title: Effect of high WDR5 expression on the hepatocellular carcinoma prognosis

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.8298

    The overall survival rates of the 113 HCC patients were compared in the high-WDR5 and low-WDR5 expression groups. Statistical significance was determined using the log-rank test. HCC, hepatocellular carcinoma; WDR5, WD repeat domain 5.
    Figure Legend Snippet: The overall survival rates of the 113 HCC patients were compared in the high-WDR5 and low-WDR5 expression groups. Statistical significance was determined using the log-rank test. HCC, hepatocellular carcinoma; WDR5, WD repeat domain 5.

    Techniques Used: Expressing

    WDR5 expression promotes HCC cell proliferation in vitro . (A) MTT assay to analyze the proliferation rate of HCC cell lines and a normal liver cell line. **P
    Figure Legend Snippet: WDR5 expression promotes HCC cell proliferation in vitro . (A) MTT assay to analyze the proliferation rate of HCC cell lines and a normal liver cell line. **P

    Techniques Used: Expressing, In Vitro, MTT Assay

    WDR5 expression was significantly elevated in HCC cell lines. (A) Reverse transcription-quantitative polymerase chain reaction was used to analyze WDR5 expression in HCC cell lines and a normal liver cell line. (B) Western blotting was used to analyze WDR5 expression in HCC cell lines and a normal liver cell line. *P
    Figure Legend Snippet: WDR5 expression was significantly elevated in HCC cell lines. (A) Reverse transcription-quantitative polymerase chain reaction was used to analyze WDR5 expression in HCC cell lines and a normal liver cell line. (B) Western blotting was used to analyze WDR5 expression in HCC cell lines and a normal liver cell line. *P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    Significantly elevated WDR5 expression levels in HCC tumor tissue compared with adjacent non-tumor tissue. (A) Reverse transcription-quantitative polymerase chain reaction and (B) western blotting were used to analyze WDR5 expression in the tumor and adjacent samples. (C) Percentages of HCC tissues and adjacent non-tumor tissues high and low WDR5 expression. Immunohistochemistry was used to analyze the WDR5 expression in (D) adjacent and (E) tumor tissue (magnification, ×200). Statistical analysis was performed with Student's t-test. **P
    Figure Legend Snippet: Significantly elevated WDR5 expression levels in HCC tumor tissue compared with adjacent non-tumor tissue. (A) Reverse transcription-quantitative polymerase chain reaction and (B) western blotting were used to analyze WDR5 expression in the tumor and adjacent samples. (C) Percentages of HCC tissues and adjacent non-tumor tissues high and low WDR5 expression. Immunohistochemistry was used to analyze the WDR5 expression in (D) adjacent and (E) tumor tissue (magnification, ×200). Statistical analysis was performed with Student's t-test. **P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry

    14) Product Images from "Human skeletal muscle derived CD133+ form functional satellite cells after intramuscular transplantation in immunodeficient host mice"

    Article Title: Human skeletal muscle derived CD133+ form functional satellite cells after intramuscular transplantation in immunodeficient host mice

    Journal: Molecular Therapy

    doi: 10.1038/mt.2014.26

    Donor-derived satellite cells are functional in vivo . ( A ) Multichannel immunostaining of human lamin A/C (red, b and g), human spectrin (red, b and g), MyoD (green, c and h), and pan-laminin (cyan, d and i) on sections of irradiated and cryodamaged muscles that had been transplanted with hCD133 + cells 1 month previously. The white arrow indicates a donor-derived satellite cell, verified by its expression of human lamin A/C (red), located outside the muscle fiber sarcolemma (spectrin + , red) but inside the basal lamina (laminin + , cyan), expressing MyoD (green), a marker of activated satellite cells. Yellow arrow points to a donor-derived myonucleus, expressing human lamin A/C (red), but located inside the muscle fiber (spectrin + , red), also expressing MyoD (green). (a–e) Bar = 25 µm, i–j are higher magnificent images taken from the same area as a–e. Bar = 5 µm. ( B ) Quantification of the number of human spectrin + fibers (blue + orange bars) and the number of recently regenerated fibers defined as small, bright neonatal myosin+ fibers (s.b. NM + , orange bars) in each individual grafted, regenerated muscles that had either not been injured by notexin (a, −NTX) or injected with notexin 7 days previously (b, +NTX). The ratio of neonatal myosin + fibers versus human spectrin + fibers (normalized) in both groups of muscles was compared using paired t -test (c). There were a significant difference between notexin treated and nontreated groups ( P = 0.0008). ( C ) Immunostaining of human lamin A/C (red, b and f), human spectrin (red, b and f), and human neonatal myosin (green, c and g) in transplanted muscles treated with notexin (upper panel, a–d) or control, noninjured (lower panel e–h). Nuclei were counterstained with DAPI (blue, a and e). Bar = 200 µm. DAPI, 4′,6-diamidino-2-phenylindole; NM, neonatal myosin; s.b. NM+, small, bright neonatal myosin+; NTX, notexin.
    Figure Legend Snippet: Donor-derived satellite cells are functional in vivo . ( A ) Multichannel immunostaining of human lamin A/C (red, b and g), human spectrin (red, b and g), MyoD (green, c and h), and pan-laminin (cyan, d and i) on sections of irradiated and cryodamaged muscles that had been transplanted with hCD133 + cells 1 month previously. The white arrow indicates a donor-derived satellite cell, verified by its expression of human lamin A/C (red), located outside the muscle fiber sarcolemma (spectrin + , red) but inside the basal lamina (laminin + , cyan), expressing MyoD (green), a marker of activated satellite cells. Yellow arrow points to a donor-derived myonucleus, expressing human lamin A/C (red), but located inside the muscle fiber (spectrin + , red), also expressing MyoD (green). (a–e) Bar = 25 µm, i–j are higher magnificent images taken from the same area as a–e. Bar = 5 µm. ( B ) Quantification of the number of human spectrin + fibers (blue + orange bars) and the number of recently regenerated fibers defined as small, bright neonatal myosin+ fibers (s.b. NM + , orange bars) in each individual grafted, regenerated muscles that had either not been injured by notexin (a, −NTX) or injected with notexin 7 days previously (b, +NTX). The ratio of neonatal myosin + fibers versus human spectrin + fibers (normalized) in both groups of muscles was compared using paired t -test (c). There were a significant difference between notexin treated and nontreated groups ( P = 0.0008). ( C ) Immunostaining of human lamin A/C (red, b and f), human spectrin (red, b and f), and human neonatal myosin (green, c and g) in transplanted muscles treated with notexin (upper panel, a–d) or control, noninjured (lower panel e–h). Nuclei were counterstained with DAPI (blue, a and e). Bar = 200 µm. DAPI, 4′,6-diamidino-2-phenylindole; NM, neonatal myosin; s.b. NM+, small, bright neonatal myosin+; NTX, notexin.

    Techniques Used: Derivative Assay, Functional Assay, In Vivo, Immunostaining, Irradiation, Expressing, Marker, Injection

    15) Product Images from "Direct intracellular signaling by the carboxy terminus of NMDA receptor GluN2 subunits regulates dendritic morphology in hippocampal CA1 pyramidal neurons"

    Article Title: Direct intracellular signaling by the carboxy terminus of NMDA receptor GluN2 subunits regulates dendritic morphology in hippocampal CA1 pyramidal neurons

    Journal: Neuroscience

    doi: 10.1016/j.neuroscience.2018.11.021

    Representative images and traces from eGFP and Golgi-Cox labeled neurons. A) eGFP labeled neurons and their corresponding traces for each genotype. B) Golgi-Cox labeled neurons and their corresponding traces for each genotype. Scale bars are equal to
    Figure Legend Snippet: Representative images and traces from eGFP and Golgi-Cox labeled neurons. A) eGFP labeled neurons and their corresponding traces for each genotype. B) Golgi-Cox labeled neurons and their corresponding traces for each genotype. Scale bars are equal to

    Techniques Used: Labeling

    16) Product Images from "(-)-Phenserine and the prevention of pre-programmed cell death and neuroinflammation in mild traumatic brain injury and Alzheimer’s disease challenged mice"

    Article Title: (-)-Phenserine and the prevention of pre-programmed cell death and neuroinflammation in mild traumatic brain injury and Alzheimer’s disease challenged mice

    Journal: Neurobiology of disease

    doi: 10.1016/j.nbd.2019.104528

    Phen mitigates mTBI-induced loss of pre- and post synaptic elements in WT and APP/PSEN1 mice. (A) Postsynaptic density protein 95 (PSD-95): mTBI induced a loss of PSD-95+ dendritic spines across all analyzed areas in WT mice, and in hippocampus of AD mice, as compared to the sham (CTRL) group. By contrast, Phen treated mTBI mice were not statistically different from the sham (CTRL) group. WT mTBI mice treated with Phen (2.5 and 5 mg/kg) possessed a greater number of PSD-95+ dendritic spines across both hippocampus and cortex, as compared to the mTBI vehicle group. Representative images showing PSD-95+ spines (green) in MAP2+ dendrites. Data are expressed as number of PSD-95+ dendritic spines/μm. (B) Synaptophysin: the total volume occupied by the presynaptic marker synaptophysin IR was evaluated across WT and APP/PSEN1 mice and found to be significantly reduced in the mTBI vehicle group, as compared to their respective sham (CTRL) group. In contrast, mTBI Phen treated mice had synaptophysin IR levels no different from sham (CTRL) mice. Importantly, Phen treatment of mTBI-challenged mice resulted in significantly higher amounts of synaptophysin IR, compared to the mTBI vehicle group, across all analyzed brain areas in both WT and AD mice. *p
    Figure Legend Snippet: Phen mitigates mTBI-induced loss of pre- and post synaptic elements in WT and APP/PSEN1 mice. (A) Postsynaptic density protein 95 (PSD-95): mTBI induced a loss of PSD-95+ dendritic spines across all analyzed areas in WT mice, and in hippocampus of AD mice, as compared to the sham (CTRL) group. By contrast, Phen treated mTBI mice were not statistically different from the sham (CTRL) group. WT mTBI mice treated with Phen (2.5 and 5 mg/kg) possessed a greater number of PSD-95+ dendritic spines across both hippocampus and cortex, as compared to the mTBI vehicle group. Representative images showing PSD-95+ spines (green) in MAP2+ dendrites. Data are expressed as number of PSD-95+ dendritic spines/μm. (B) Synaptophysin: the total volume occupied by the presynaptic marker synaptophysin IR was evaluated across WT and APP/PSEN1 mice and found to be significantly reduced in the mTBI vehicle group, as compared to their respective sham (CTRL) group. In contrast, mTBI Phen treated mice had synaptophysin IR levels no different from sham (CTRL) mice. Importantly, Phen treatment of mTBI-challenged mice resulted in significantly higher amounts of synaptophysin IR, compared to the mTBI vehicle group, across all analyzed brain areas in both WT and AD mice. *p

    Techniques Used: Mouse Assay, Marker

    17) Product Images from "Semaphorin-5B Controls Spiral Ganglion Neuron Branch Refinement during Development"

    Article Title: Semaphorin-5B Controls Spiral Ganglion Neuron Branch Refinement during Development

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0113-19.2019

    SGNs undergo a stereotyped refinement processes while Sema5b and Plxnas are expressed in differentiating hair cells and SGNs, respectively. A , A whole-mount view of hair cells from a P1 cochlea, shown by MyoVI labeling, organized into one row of inner hair cells (IHCs) and three rows of outer hair cells (OHCs). B , Sample from A with NF200 labeling of type I and II SGNs. C , A cross-sectional view of the same structures and labeling from the orientation of the dotted line in B . D , At E15.5, immature SGN peripheral axons have many terminal branches (red arrowheads), which explore the differentiating cochlear sensory domain. A few days before birth, SGNs begin to retract and remove excess branches, as type-I SGNs prepare to ultimately form a single connection with a single IHC. E – G , In situ hybridizations showing that Sema5b mRNA is specifically expressed in hair cells, identified by arrows, in late embryonic ( E ) and early postnatal ( F ) development, but is largely eliminated by P5 ( G ). H – K , Plxna mRNA expression in E17.5 cochlear cross-sections using Plxna1 ( H , I ) and Plxna3 ( J , K ) antisense probes along with sense controls. Plxna1 transcripts are visible in the spiral ganglion (SG) and in lateral region of the cochlear sensory domain (arrowhead) in H . Plxna3 transcripts are visible in the SG in J . Sense controls show no reactivity. Scale bar (in G ): A , B , ∼100 μm; C , 20 μm; E – G , 60 μm; H – K , 200 μm.
    Figure Legend Snippet: SGNs undergo a stereotyped refinement processes while Sema5b and Plxnas are expressed in differentiating hair cells and SGNs, respectively. A , A whole-mount view of hair cells from a P1 cochlea, shown by MyoVI labeling, organized into one row of inner hair cells (IHCs) and three rows of outer hair cells (OHCs). B , Sample from A with NF200 labeling of type I and II SGNs. C , A cross-sectional view of the same structures and labeling from the orientation of the dotted line in B . D , At E15.5, immature SGN peripheral axons have many terminal branches (red arrowheads), which explore the differentiating cochlear sensory domain. A few days before birth, SGNs begin to retract and remove excess branches, as type-I SGNs prepare to ultimately form a single connection with a single IHC. E – G , In situ hybridizations showing that Sema5b mRNA is specifically expressed in hair cells, identified by arrows, in late embryonic ( E ) and early postnatal ( F ) development, but is largely eliminated by P5 ( G ). H – K , Plxna mRNA expression in E17.5 cochlear cross-sections using Plxna1 ( H , I ) and Plxna3 ( J , K ) antisense probes along with sense controls. Plxna1 transcripts are visible in the spiral ganglion (SG) and in lateral region of the cochlear sensory domain (arrowhead) in H . Plxna3 transcripts are visible in the SG in J . Sense controls show no reactivity. Scale bar (in G ): A , B , ∼100 μm; C , 20 μm; E – G , 60 μm; H – K , 200 μm.

    Techniques Used: Labeling, Immunohistochemistry, In Situ, Expressing

    18) Product Images from "Early Loss of Vision Results in Extensive Reorganization of Plasticity-Related Receptors and Alterations in Hippocampal Function That Extend Through Adulthood"

    Article Title: Early Loss of Vision Results in Extensive Reorganization of Plasticity-Related Receptors and Alterations in Hippocampal Function That Extend Through Adulthood

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhy297

    GluN2B expression is globally increased in CBA/J mice 4 months postnatally. ( A ) Bar charts represent mean GluN2B receptor optical densities in different brain regions 4 months postnatally in CBA/J mice compared with (CBA/CaOlaHsd) controls. Receptor density is significantly increased in the piriform cortex (PiC), somatosensory cortex (SC), posterior parietal cortex (PPC), visual cortex (VC), auditory cortex (AuC), dentate gyrus (DG), CA1, CA3, and CA4. Data are means ± SEM. * P
    Figure Legend Snippet: GluN2B expression is globally increased in CBA/J mice 4 months postnatally. ( A ) Bar charts represent mean GluN2B receptor optical densities in different brain regions 4 months postnatally in CBA/J mice compared with (CBA/CaOlaHsd) controls. Receptor density is significantly increased in the piriform cortex (PiC), somatosensory cortex (SC), posterior parietal cortex (PPC), visual cortex (VC), auditory cortex (AuC), dentate gyrus (DG), CA1, CA3, and CA4. Data are means ± SEM. * P

    Techniques Used: Expressing, Crocin Bleaching Assay, Mouse Assay

    19) Product Images from "Newly developed TGF-?2 knock down transgenic mouse lines express TGF-?2 differently and its distribution in multiple tissues varies"

    Article Title: Newly developed TGF-?2 knock down transgenic mouse lines express TGF-?2 differently and its distribution in multiple tissues varies

    Journal: BMC Biochemistry

    doi: 10.1186/1471-2091-14-21

    Protein expressions of TGF-β2 detected by WB in different tissues. Figure 2 Lane 1–5, TGF-β2 protein expression; Lane 1, WT; Lane 2, Founder 66; Lane 3, Founder 16; Lane 4, Founder 53; Lane 5, Founder 41. A , a : olfactory bulb; B , b : cortex; C , c : frontal lobe; D , d : basal forebrain; E , e : cerebellum; F , f : hypothalamus; G , g : medulla oblongata; H , h : spinal cord; I , i : trachea; J , j : lung; K , k : heart; L , l : liver; M , m : spleen; N , n : kidney; O , o : adrenal gland; P , p : intestines; Q , q : skeletal muscles; R , r : epidermis. Beta-tubulin was chased as the control.
    Figure Legend Snippet: Protein expressions of TGF-β2 detected by WB in different tissues. Figure 2 Lane 1–5, TGF-β2 protein expression; Lane 1, WT; Lane 2, Founder 66; Lane 3, Founder 16; Lane 4, Founder 53; Lane 5, Founder 41. A , a : olfactory bulb; B , b : cortex; C , c : frontal lobe; D , d : basal forebrain; E , e : cerebellum; F , f : hypothalamus; G , g : medulla oblongata; H , h : spinal cord; I , i : trachea; J , j : lung; K , k : heart; L , l : liver; M , m : spleen; N , n : kidney; O , o : adrenal gland; P , p : intestines; Q , q : skeletal muscles; R , r : epidermis. Beta-tubulin was chased as the control.

    Techniques Used: Western Blot, Expressing

    Recombination plasmid for Tg mice with TGF-β2 down-regulation. A : showed the schedules of recombination plasmid for pcDNA6.2-GW/EmGFP-miR of TGF-β2 gene silence, which composed with 5699 nucleotides. The 293T cells were transfected with the transgenic vectors (pcDNA3.1 (+) of pcDNA6.2-GW/EmGFP-miR-TGF-β1). RT-PCR was employed to evaluate the effects of PDGF-BB down-regulation transformants. B : showed the amplification plot of RT-PCR. Red arrows showed the selected cell lines as they had the lowest levels. Black arrow indicated the control ones. C : shows the represented bands of semi-quantity PCR products electrophoresed in 1% agarose gel stained with EB. Lane 1: DL2000 DNA Marker (from up to down: 2000 bp, 1000 bp, 750 bp, 500 bp, 250 bp, 100 bp respectively); Lane 2–4: 293T cells transfected with silence expression vector for TGF-β2 gene (lane 3: NO.21). Arrows in Figure 5 C revealed the target transformants (NO.21) for TGF-β2 expressional silence as they had the lowest levels.
    Figure Legend Snippet: Recombination plasmid for Tg mice with TGF-β2 down-regulation. A : showed the schedules of recombination plasmid for pcDNA6.2-GW/EmGFP-miR of TGF-β2 gene silence, which composed with 5699 nucleotides. The 293T cells were transfected with the transgenic vectors (pcDNA3.1 (+) of pcDNA6.2-GW/EmGFP-miR-TGF-β1). RT-PCR was employed to evaluate the effects of PDGF-BB down-regulation transformants. B : showed the amplification plot of RT-PCR. Red arrows showed the selected cell lines as they had the lowest levels. Black arrow indicated the control ones. C : shows the represented bands of semi-quantity PCR products electrophoresed in 1% agarose gel stained with EB. Lane 1: DL2000 DNA Marker (from up to down: 2000 bp, 1000 bp, 750 bp, 500 bp, 250 bp, 100 bp respectively); Lane 2–4: 293T cells transfected with silence expression vector for TGF-β2 gene (lane 3: NO.21). Arrows in Figure 5 C revealed the target transformants (NO.21) for TGF-β2 expressional silence as they had the lowest levels.

    Techniques Used: Plasmid Preparation, Mouse Assay, Transfection, Transgenic Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Marker, Expressing

    Genotypes detection for the TGF-β2-kd Tg mice. The positive Tg mice detected by PCR. Figure 1 showed the representative lanes of products electrophoresed in 1% agarose gel stained with EB. Lane 1: DNA Marker DL 2,000 (from up to down: 2000 bp, 1000 bp, 750 bp, 500 bp, 250 bp, 100 bp respectively). Lane 2–9: The PCR productions of inserted fragment from different heterozygous transgenic offspring of TGF-β2-kd lines. Lane 2, Lane 3, Lane 6 and Lane 7: WT; Lane 4: Founder 66; Lane 5: Founder 16; Lane 8: Founder 53; Lane 9: Founder 41.
    Figure Legend Snippet: Genotypes detection for the TGF-β2-kd Tg mice. The positive Tg mice detected by PCR. Figure 1 showed the representative lanes of products electrophoresed in 1% agarose gel stained with EB. Lane 1: DNA Marker DL 2,000 (from up to down: 2000 bp, 1000 bp, 750 bp, 500 bp, 250 bp, 100 bp respectively). Lane 2–9: The PCR productions of inserted fragment from different heterozygous transgenic offspring of TGF-β2-kd lines. Lane 2, Lane 3, Lane 6 and Lane 7: WT; Lane 4: Founder 66; Lane 5: Founder 16; Lane 8: Founder 53; Lane 9: Founder 41.

    Techniques Used: Mouse Assay, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Marker, Transgenic Assay

    Relative expressions of TGF-β2 in different tissues of Tg mice. Figure 3 showed the relative optical density (O.D.) of TGF-β2 protein levels in multiple tissues of Tg mice and that of WT ones (n = 6). Values plotted are means ± SD. * compared with WT, P
    Figure Legend Snippet: Relative expressions of TGF-β2 in different tissues of Tg mice. Figure 3 showed the relative optical density (O.D.) of TGF-β2 protein levels in multiple tissues of Tg mice and that of WT ones (n = 6). Values plotted are means ± SD. * compared with WT, P

    Techniques Used: Mouse Assay

    Locations of TGF-β2 in multiply tissues of Tg mice. The arrow showed the representative IR of TGF-β2 in Founder 66. A : olfactory bulb (arrow showed supporting cell); B : cortex frontal lobe (arrows showed neuron); C : basal forebrain (arrows showed neuron); D : hypothalamus (arrows showed neuron); E , F and G : spinal cord (arrows showed, E and F : motor neurons in the ventral horn); G: astrocyte-like cells in whiter matter); H : lung (arrows showed the lung epithelial cell); I and J : heart (arrows showed the sarcolemma); K : liver (arrows showed); L : spleen (arrow showed subendothelial smooth muscle cell); M : kidney (arrows showed epithelial cells); N : adrenal gland; O : intestines (arrows showed the staining lamina); P and Q : skeletal muscles (arrows showed the staining sarcolemma); R : epidermis (arrows showed the basal cells); S : control of olfactory bulb; T : control of brain. Magnifications: C , D , N , Q and R : 400×; other: 200×; Scale bar: 10 μm.
    Figure Legend Snippet: Locations of TGF-β2 in multiply tissues of Tg mice. The arrow showed the representative IR of TGF-β2 in Founder 66. A : olfactory bulb (arrow showed supporting cell); B : cortex frontal lobe (arrows showed neuron); C : basal forebrain (arrows showed neuron); D : hypothalamus (arrows showed neuron); E , F and G : spinal cord (arrows showed, E and F : motor neurons in the ventral horn); G: astrocyte-like cells in whiter matter); H : lung (arrows showed the lung epithelial cell); I and J : heart (arrows showed the sarcolemma); K : liver (arrows showed); L : spleen (arrow showed subendothelial smooth muscle cell); M : kidney (arrows showed epithelial cells); N : adrenal gland; O : intestines (arrows showed the staining lamina); P and Q : skeletal muscles (arrows showed the staining sarcolemma); R : epidermis (arrows showed the basal cells); S : control of olfactory bulb; T : control of brain. Magnifications: C , D , N , Q and R : 400×; other: 200×; Scale bar: 10 μm.

    Techniques Used: Mouse Assay, Staining

    20) Product Images from "Hypothermia and Pharmacological Regimens that Prevent Overexpression and Overactivity of the Extracellular Calcium-Sensing Receptor Protect Neurons against Traumatic Brain Injury"

    Article Title: Hypothermia and Pharmacological Regimens that Prevent Overexpression and Overactivity of the Extracellular Calcium-Sensing Receptor Protect Neurons against Traumatic Brain Injury

    Journal: Journal of Neurotrauma

    doi: 10.1089/neu.2012.2691

    Effects of controlled cortical impact (CCI) and hypothermia on the expression of calcium-sensing receptor (CaSR) and GABA-B-R1 protein in neurons at impact sites. Immunohistological detection of (a) CaSR and (b) GABA-B-R1 in cortical neurons in mice subjected to CCI with hypothermia (CCI:33°C, n =3 mice) or normothermia (CCI:37°C, n =3 mice) vs. sham control mice with hypothermia (sham:33°C, n =3 mice) or normothermia (CCI:37°C, n =3 mice). Black boxes contain images of higher-power view at corresponding brain regions. Brown diamino benzidine stain depicts immunoreactivity in the brain sections counterstained with hematoxylin. Bar=50 μm.
    Figure Legend Snippet: Effects of controlled cortical impact (CCI) and hypothermia on the expression of calcium-sensing receptor (CaSR) and GABA-B-R1 protein in neurons at impact sites. Immunohistological detection of (a) CaSR and (b) GABA-B-R1 in cortical neurons in mice subjected to CCI with hypothermia (CCI:33°C, n =3 mice) or normothermia (CCI:37°C, n =3 mice) vs. sham control mice with hypothermia (sham:33°C, n =3 mice) or normothermia (CCI:37°C, n =3 mice). Black boxes contain images of higher-power view at corresponding brain regions. Brown diamino benzidine stain depicts immunoreactivity in the brain sections counterstained with hematoxylin. Bar=50 μm.

    Techniques Used: Expressing, Mouse Assay, Staining

    21) Product Images from "The Anticancer Effect of (1S,2S,3E,7E,11E)-3,7,11,15-Cembratetraen-17,2-olide(LS-1) through the Activation of TGF-β Signaling in SNU-C5/5-FU, Fluorouracil-Resistant Human Colon Cancer Cells"

    Article Title: The Anticancer Effect of (1S,2S,3E,7E,11E)-3,7,11,15-Cembratetraen-17,2-olide(LS-1) through the Activation of TGF-β Signaling in SNU-C5/5-FU, Fluorouracil-Resistant Human Colon Cancer Cells

    Journal: Marine Drugs

    doi: 10.3390/md13031340

    The effect of LS-1 on the interaction between TGFβRI and CEA in SNU-C5/5-FU. ( A ) SNU-C5/5-FU were treated with 7.1 μM of LS-1 for 12, 24 and 48 h. The interaction between TGFβRI and CEA was examined by immunoprecipitation with anti-TGFβRI antibody and with immunoblotting with anti-TGFβRI antibody and anti-CEA antibody; ( B ) Data represent the percentage of CEA expression in SNU-C5/5-FU. The data are presented as the mean value ± SD from three independent trials. * p
    Figure Legend Snippet: The effect of LS-1 on the interaction between TGFβRI and CEA in SNU-C5/5-FU. ( A ) SNU-C5/5-FU were treated with 7.1 μM of LS-1 for 12, 24 and 48 h. The interaction between TGFβRI and CEA was examined by immunoprecipitation with anti-TGFβRI antibody and with immunoblotting with anti-TGFβRI antibody and anti-CEA antibody; ( B ) Data represent the percentage of CEA expression in SNU-C5/5-FU. The data are presented as the mean value ± SD from three independent trials. * p

    Techniques Used: Immunoprecipitation, Expressing

    22) Product Images from "Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles"

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    Journal: Viruses

    doi: 10.3390/v10100575

    The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.
    Figure Legend Snippet: The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Techniques Used: Marker, Transfection, Mutagenesis, Variant Assay, Microscopy

    Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.
    Figure Legend Snippet: Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Incubation, Microscopy

    Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.
    Figure Legend Snippet: Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Techniques Used: Transfection, Marker, Microscopy

    23) Product Images from "Human dendritic cell-specific ICAM-3-grabbing non-integrin downstream signaling alleviates renal fibrosis via Raf-1 activation in systemic candidiasis"

    Article Title: Human dendritic cell-specific ICAM-3-grabbing non-integrin downstream signaling alleviates renal fibrosis via Raf-1 activation in systemic candidiasis

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/s41423-018-0161-5

    Renal proximal tubular epithelial cells in hDC-SIGN transgenic mice express hDC-SIGN. a Map of the pK14TyrPolII-hDC-SIGNpA-IN2B vector. The human DC-SIGN (hDC-SIGN) gene was cloned under the control of the murine polymerase II large subunit gene promoter, followed by poly(A) tail and two copies of HS4 insulator genes. b , c Kidneys from hDC-SIGN transgenic and littermate control mice were collected after perfusion with PBS. b Paraffin-embedded kidney sections were stained for hDC-SIGN (brown). Slides were counterstained with hematoxylin (blue). Original magnification, ×200. Scale bar = 100 µm. c Cryosections of kidneys were stained for hDC-SIGN (orange), DBA ( Dolichos biflorus agglutinin, red), LTL ( Lotus tetragonolobus lectin, green), and nuclei (Hoechst 33258, blue). DBA and LTL mark the distal and proximal tubules in the renal cortex, respectively. White arrows point to hDC-SIGN + cells. Slides were read under a confocal microscope. Original magnification, ×400. Scale bar = 20 µm
    Figure Legend Snippet: Renal proximal tubular epithelial cells in hDC-SIGN transgenic mice express hDC-SIGN. a Map of the pK14TyrPolII-hDC-SIGNpA-IN2B vector. The human DC-SIGN (hDC-SIGN) gene was cloned under the control of the murine polymerase II large subunit gene promoter, followed by poly(A) tail and two copies of HS4 insulator genes. b , c Kidneys from hDC-SIGN transgenic and littermate control mice were collected after perfusion with PBS. b Paraffin-embedded kidney sections were stained for hDC-SIGN (brown). Slides were counterstained with hematoxylin (blue). Original magnification, ×200. Scale bar = 100 µm. c Cryosections of kidneys were stained for hDC-SIGN (orange), DBA ( Dolichos biflorus agglutinin, red), LTL ( Lotus tetragonolobus lectin, green), and nuclei (Hoechst 33258, blue). DBA and LTL mark the distal and proximal tubules in the renal cortex, respectively. White arrows point to hDC-SIGN + cells. Slides were read under a confocal microscope. Original magnification, ×400. Scale bar = 20 µm

    Techniques Used: Transgenic Assay, Mouse Assay, Plasmid Preparation, Clone Assay, Staining, Microscopy

    24) Product Images from "Inflammatory Responses Are Not Sufficient to Cause Delayed Neuronal Death in ATP-Induced Acute Brain Injury"

    Article Title: Inflammatory Responses Are Not Sufficient to Cause Delayed Neuronal Death in ATP-Induced Acute Brain Injury

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0013756

    Death of dopaminergic neurons and microglia in the SNpc induced by ATP. ( A ) ATP (100 nmol in 2 µl PBS) or PBS (2 µl) was unilaterally injected into SNpc (*, injection sites), and brains were obtained after 3 h. Brain sections (30 µm thickness) of the midbrain including the entire SN were prepared, every sixth serial section selected and stained with TH (upper panel) and Iba-1 (lower panel) antibodies, and visualized with biotin-conjugated secondary antibodies and enzymatic detection with the avidin/biotin system unless indicated. At 100 nmol, mild neuronal and microglial damage occurred, thus, 100 nmol ATP was employed for in vivo experiments in this study. Photographs of the most damaged sections were obtained. The contralateral side (contra) and PBS-injected rat brain sections were used as control. ( B ) Serial sections obtained at 3 h were labeled with TH (left panel), Iba-1 (middle panel), and TH/Iba-1 (right panel) antibodies. For visualization of the double-labeling, color reactions using DAB (for TH) and DAB/nickel sulfate (for Iba-1) were applied. Dotted lines indicated damage areas. ( C ) Brain tissue obtained 3 h post ATP treatment was subjected to electron microscopy, as described in “ Materials and Methods ”. Nuclei of neuron (N, white arrow), astrocytes (A, white arrowhead), and microglia (M, black arrow) were shown in intact rat brain whereas cellular structures were severely disrupted in ATP-injected brain. Data in this study are representative of at least 5 animals. Scale bars, 200 µm (A); 100 µm (B); 5 µm (C).
    Figure Legend Snippet: Death of dopaminergic neurons and microglia in the SNpc induced by ATP. ( A ) ATP (100 nmol in 2 µl PBS) or PBS (2 µl) was unilaterally injected into SNpc (*, injection sites), and brains were obtained after 3 h. Brain sections (30 µm thickness) of the midbrain including the entire SN were prepared, every sixth serial section selected and stained with TH (upper panel) and Iba-1 (lower panel) antibodies, and visualized with biotin-conjugated secondary antibodies and enzymatic detection with the avidin/biotin system unless indicated. At 100 nmol, mild neuronal and microglial damage occurred, thus, 100 nmol ATP was employed for in vivo experiments in this study. Photographs of the most damaged sections were obtained. The contralateral side (contra) and PBS-injected rat brain sections were used as control. ( B ) Serial sections obtained at 3 h were labeled with TH (left panel), Iba-1 (middle panel), and TH/Iba-1 (right panel) antibodies. For visualization of the double-labeling, color reactions using DAB (for TH) and DAB/nickel sulfate (for Iba-1) were applied. Dotted lines indicated damage areas. ( C ) Brain tissue obtained 3 h post ATP treatment was subjected to electron microscopy, as described in “ Materials and Methods ”. Nuclei of neuron (N, white arrow), astrocytes (A, white arrowhead), and microglia (M, black arrow) were shown in intact rat brain whereas cellular structures were severely disrupted in ATP-injected brain. Data in this study are representative of at least 5 animals. Scale bars, 200 µm (A); 100 µm (B); 5 µm (C).

    Techniques Used: Injection, Staining, Avidin-Biotin Assay, In Vivo, Labeling, Electron Microscopy

    25) Product Images from "Simple buffers for 3D STORM microscopy"

    Article Title: Simple buffers for 3D STORM microscopy

    Journal: Biomedical Optics Express

    doi: 10.1364/BOE.4.000885

    STORM imaging of microtubules (see section 2.2 for more details) in Vectashield. (A): Widefield image (B): Single frame, (C1): Reconstructed STORM image, with blow-up on the ROI in (C2). scale-bar = 5 μ m for (A),(B),(C1) and 1 μ m for C2.
    Figure Legend Snippet: STORM imaging of microtubules (see section 2.2 for more details) in Vectashield. (A): Widefield image (B): Single frame, (C1): Reconstructed STORM image, with blow-up on the ROI in (C2). scale-bar = 5 μ m for (A),(B),(C1) and 1 μ m for C2.

    Techniques Used: Imaging

    STORM imaging of Alexa-647 stained microtubules in a Vectashield/TRIS-Glycerol mixture: (A) 50% Vectashield and (B) 25% Vectashield. The different panels represent: (1) STORM image reconstructed from 15.000 frames, scale-bar = 500 nm (2) photon count distribution per frame and per molecule, averaged over three data-sets, and (3) standard deviation of multiple localizations giving a measure of the frame localization precision.
    Figure Legend Snippet: STORM imaging of Alexa-647 stained microtubules in a Vectashield/TRIS-Glycerol mixture: (A) 50% Vectashield and (B) 25% Vectashield. The different panels represent: (1) STORM image reconstructed from 15.000 frames, scale-bar = 500 nm (2) photon count distribution per frame and per molecule, averaged over three data-sets, and (3) standard deviation of multiple localizations giving a measure of the frame localization precision.

    Techniques Used: Imaging, Staining, Standard Deviation

    (A) Absorption spectrum of Vectashield, as well as normalized emission spectra measured at 3 different wavelengths: 400 nm (B), 560 nm (C) and 630 nm (D) with normalization factor indicated in the top right corner.
    Figure Legend Snippet: (A) Absorption spectrum of Vectashield, as well as normalized emission spectra measured at 3 different wavelengths: 400 nm (B), 560 nm (C) and 630 nm (D) with normalization factor indicated in the top right corner.

    Techniques Used:

    Statistics on STORM imaging performed in 25% Vectashield - 75% TRIS-Glycerol in which were added 1% NPG (w/v) (A), 20 mM DABCO (B), and 10 mM Lipoic Acid (C). The different panels represent: (1) photon count distribution per frame and per molecule, averaged over three datasets, (2) standard deviation of multiple localizations giving a measure of the frame localization precision, and (3) Density of molecules as a function of number of recorded frames, averaged over three measurements, with error bars indicating the standard deviation.
    Figure Legend Snippet: Statistics on STORM imaging performed in 25% Vectashield - 75% TRIS-Glycerol in which were added 1% NPG (w/v) (A), 20 mM DABCO (B), and 10 mM Lipoic Acid (C). The different panels represent: (1) photon count distribution per frame and per molecule, averaged over three datasets, (2) standard deviation of multiple localizations giving a measure of the frame localization precision, and (3) Density of molecules as a function of number of recorded frames, averaged over three measurements, with error bars indicating the standard deviation.

    Techniques Used: Imaging, Standard Deviation

    Quantifying the quality of Vectashield as a STORM buffer for Alexa-647: (A) photon count distribution per frame (blue) and per molecule (red), obtained by grouping consecutive frame localizations and (B) standard deviation of multiple localizations (see section 2.3 for grouping details), with mean values displayed in the top right corner (C) Density of molecules as a function of number of recorded frames, averaged over three measurements. The error bar indicates the standard deviation. (D) STORM image of microtubule, on which the hollowness of the structure can be resolved, as quantified in the profile taken over the 200 nm yellow-boxed region with ≈ 35 nm between the two peaks, consistent with a 25 mm structure broadened by the antibodies.
    Figure Legend Snippet: Quantifying the quality of Vectashield as a STORM buffer for Alexa-647: (A) photon count distribution per frame (blue) and per molecule (red), obtained by grouping consecutive frame localizations and (B) standard deviation of multiple localizations (see section 2.3 for grouping details), with mean values displayed in the top right corner (C) Density of molecules as a function of number of recorded frames, averaged over three measurements. The error bar indicates the standard deviation. (D) STORM image of microtubule, on which the hollowness of the structure can be resolved, as quantified in the profile taken over the 200 nm yellow-boxed region with ≈ 35 nm between the two peaks, consistent with a 25 mm structure broadened by the antibodies.

    Techniques Used: Standard Deviation

    STORM images obtained with the other working dyes (A) Alexa-555 in 20% Vectashield-80% TRIS-Glycerol (B) Cy-5 (C) CF-647 (D) Alexa-700, all in pure Vec-tashield. scale-bar = 5 μ m.
    Figure Legend Snippet: STORM images obtained with the other working dyes (A) Alexa-555 in 20% Vectashield-80% TRIS-Glycerol (B) Cy-5 (C) CF-647 (D) Alexa-700, all in pure Vec-tashield. scale-bar = 5 μ m.

    Techniques Used:

    (A) STORM image of CEP-152 stained with Cy3 using a buffer 40% Vectashield + 1% NPG + 20 mM DABCO, which improves the quality of Cy3 blinking. Scale-bar = 500 nm (B) Radial intensity distribution measured from the yellow ROI defined in (A), and Lorentzian fit showing a peak at r = 143 nm.
    Figure Legend Snippet: (A) STORM image of CEP-152 stained with Cy3 using a buffer 40% Vectashield + 1% NPG + 20 mM DABCO, which improves the quality of Cy3 blinking. Scale-bar = 500 nm (B) Radial intensity distribution measured from the yellow ROI defined in (A), and Lorentzian fit showing a peak at r = 143 nm.

    Techniques Used: Staining

    (A) Index matching with Vectashield: Optical index as a function of Vectashield concentration starting from PBS (red) or TDE (blue), and imaging performed at n = 1.5 (adapted to oil objectives) and n=1.4 (adapted to glycerol objectives) (B–D) STORM imaging of microtubules immunostained with Alexa-647 for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively, scale-bar = 500 nm (C–E): photon count distribution per frame and per molecule, averaged over three datasets for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively.
    Figure Legend Snippet: (A) Index matching with Vectashield: Optical index as a function of Vectashield concentration starting from PBS (red) or TDE (blue), and imaging performed at n = 1.5 (adapted to oil objectives) and n=1.4 (adapted to glycerol objectives) (B–D) STORM imaging of microtubules immunostained with Alexa-647 for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively, scale-bar = 500 nm (C–E): photon count distribution per frame and per molecule, averaged over three datasets for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively.

    Techniques Used: Concentration Assay, Imaging

    3D STORM of Alexa-647-labeled microtubules in Vectashield: (A) Imaging performed in 25% Vectashield-75 % TRIS-Glycerol, scale-bar = 5 μ m. (B1 2): axial profile taken from the two regions delimited in A (yellow for (B1), showing a single microtubule; red for (B2) showing two well-resolved microtubules crossing at a distance of ≈ 160 nm).
    Figure Legend Snippet: 3D STORM of Alexa-647-labeled microtubules in Vectashield: (A) Imaging performed in 25% Vectashield-75 % TRIS-Glycerol, scale-bar = 5 μ m. (B1 2): axial profile taken from the two regions delimited in A (yellow for (B1), showing a single microtubule; red for (B2) showing two well-resolved microtubules crossing at a distance of ≈ 160 nm).

    Techniques Used: Labeling, Imaging

    26) Product Images from "Motoneuron Wnts regulate neuromuscular junction development"

    Article Title: Motoneuron Wnts regulate neuromuscular junction development

    Journal: eLife

    doi: 10.7554/eLife.34625

    Wls loss in motoneuron does not enhance synapse number in single muscle fiber. ( A ) Representative images of individual gastrocnemius muscle fibers of 2-month-old control and HB9-Wls -/- mice. To visualize AChR clusters on single muscle fibers, gastrocnemius was fixed in 4% PFA and stained with R-BTX (Red) and DAPI (Blue) to show AChR clusters and myonuclei. Muscles were washed three times in PBS and teased into single fibers and mounted in Vectashield mounting medium. We counted the number of AChR cluster in each single muscle fiber and defined that synapse elimination was impaired when more than one AChR cluster was found after P15. ( B ) Quantitative analysis of NMJ number per muscle fiber. NMJ number per muscle fiber was comparable between control and HB9-Wls -/- mice at P0, P7, P15, and P60. n = 3 mice per group. Unpaired t-test, p > 0.05.
    Figure Legend Snippet: Wls loss in motoneuron does not enhance synapse number in single muscle fiber. ( A ) Representative images of individual gastrocnemius muscle fibers of 2-month-old control and HB9-Wls -/- mice. To visualize AChR clusters on single muscle fibers, gastrocnemius was fixed in 4% PFA and stained with R-BTX (Red) and DAPI (Blue) to show AChR clusters and myonuclei. Muscles were washed three times in PBS and teased into single fibers and mounted in Vectashield mounting medium. We counted the number of AChR cluster in each single muscle fiber and defined that synapse elimination was impaired when more than one AChR cluster was found after P15. ( B ) Quantitative analysis of NMJ number per muscle fiber. NMJ number per muscle fiber was comparable between control and HB9-Wls -/- mice at P0, P7, P15, and P60. n = 3 mice per group. Unpaired t-test, p > 0.05.

    Techniques Used: Mouse Assay, Staining

    27) Product Images from "Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles"

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    Journal: Viruses

    doi: 10.3390/v10100575

    The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.
    Figure Legend Snippet: The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Techniques Used: Marker, Transfection, Mutagenesis, Variant Assay, Microscopy

    Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.
    Figure Legend Snippet: Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Incubation, Microscopy

    Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.
    Figure Legend Snippet: Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Techniques Used: Transfection, Marker, Microscopy

    28) Product Images from "Helicase LSH/Hells regulates kinetochore function, histone H3/Thr3 phosphorylation and centromere transcription during oocyte meiosis"

    Article Title: Helicase LSH/Hells regulates kinetochore function, histone H3/Thr3 phosphorylation and centromere transcription during oocyte meiosis

    Journal: Nature Communications

    doi: 10.1038/s41467-020-18009-3

    LSH knockout in preovulatory oocytes induces increased H3T3 phosphorylation and abnormal kinetochore structure. a Wild-type (WT) oocyte showing H3T3ph localization (red) to the inter-chromatid axis of metaphase-I chromosome bivalents (arrow), fused sister kinetochores are stained with CREST (green). DNA was counterstained with DAPI (blue) scale bar = 5 μm. Oocytes from Lsh -cKO mutant females exhibit a striking increase in the levels of H3T3ph (arrow) decorating the entire chromosome. Higher levels of H3T3ph were confirmed by western blot analysis and result from a significant increase in the proportion of oocytes with elevated H3T3ph at metaphase-I and metaphase-II stage (bar plot; P = 0.0063). Fluorescence intensity at M-I and M-II (scatter plots; P = 0.0012 and P
    Figure Legend Snippet: LSH knockout in preovulatory oocytes induces increased H3T3 phosphorylation and abnormal kinetochore structure. a Wild-type (WT) oocyte showing H3T3ph localization (red) to the inter-chromatid axis of metaphase-I chromosome bivalents (arrow), fused sister kinetochores are stained with CREST (green). DNA was counterstained with DAPI (blue) scale bar = 5 μm. Oocytes from Lsh -cKO mutant females exhibit a striking increase in the levels of H3T3ph (arrow) decorating the entire chromosome. Higher levels of H3T3ph were confirmed by western blot analysis and result from a significant increase in the proportion of oocytes with elevated H3T3ph at metaphase-I and metaphase-II stage (bar plot; P = 0.0063). Fluorescence intensity at M-I and M-II (scatter plots; P = 0.0012 and P

    Techniques Used: Knock-Out, Staining, Mutagenesis, Western Blot, Fluorescence

    29) Product Images from "Reorganization of Basolateral Amygdala-Subiculum Circuitry in Mouse Epilepsy Model"

    Article Title: Reorganization of Basolateral Amygdala-Subiculum Circuitry in Mouse Epilepsy Model

    Journal: Frontiers in Neuroanatomy

    doi: 10.3389/fnana.2015.00167

    Increased terminal-like structures in the vSub of epileptic mice. Representative images showing PHA-L-immunopositive fibers in the vSub region of control (A,C) or epileptic (B,D) mice. Graph shows density of PHA-L-labeled terminal-like structures in both groups of mice normalized with area shrinkage in the SE2m group (E) (Student’s t -test, * p
    Figure Legend Snippet: Increased terminal-like structures in the vSub of epileptic mice. Representative images showing PHA-L-immunopositive fibers in the vSub region of control (A,C) or epileptic (B,D) mice. Graph shows density of PHA-L-labeled terminal-like structures in both groups of mice normalized with area shrinkage in the SE2m group (E) (Student’s t -test, * p

    Techniques Used: Mouse Assay, Labeling

    30) Product Images from "TGF-β activates APC through Cdh1 binding for Cks1 and Skp2 proteasomal destruction stabilizing p27kip1 for normal endometrial growth"

    Article Title: TGF-β activates APC through Cdh1 binding for Cks1 and Skp2 proteasomal destruction stabilizing p27kip1 for normal endometrial growth

    Journal: Cell Cycle

    doi: 10.1080/15384101.2016.1150393

    TGF-β increases Cdh1 and decreases Skp2 and Cks1 to stabilize nuclear p27 in primary endometrial epithelial cells (EECs). (A) TGF-β effect on the protein levels of Cdh1, p27, Skp2, and Cks1. Normal primary EECs isolated from proliferative
    Figure Legend Snippet: TGF-β increases Cdh1 and decreases Skp2 and Cks1 to stabilize nuclear p27 in primary endometrial epithelial cells (EECs). (A) TGF-β effect on the protein levels of Cdh1, p27, Skp2, and Cks1. Normal primary EECs isolated from proliferative

    Techniques Used: Isolation

    Immunohistochemical analysis of normal, hyperplastic, and type I endometrial (endometrioid) cancer tissues shows inverse expression of p27 and Cks1. (A) Tissues from normal proliferative (PE; panels a-b) and secretory (SE; c-d) endometrium, endometrial
    Figure Legend Snippet: Immunohistochemical analysis of normal, hyperplastic, and type I endometrial (endometrioid) cancer tissues shows inverse expression of p27 and Cks1. (A) Tissues from normal proliferative (PE; panels a-b) and secretory (SE; c-d) endometrium, endometrial

    Techniques Used: Immunohistochemistry, Expressing

    Cdh1 is required for TGF-β-mediated inhibition of proliferation and stabilization of nuclear p27. (A) Knocking-down Cdh1 blocks TGF-β induction of p27 and increases Skp2 and Cks1. HEC-1A cells were transiently transfected with Cdh1 siRNA
    Figure Legend Snippet: Cdh1 is required for TGF-β-mediated inhibition of proliferation and stabilization of nuclear p27. (A) Knocking-down Cdh1 blocks TGF-β induction of p27 and increases Skp2 and Cks1. HEC-1A cells were transiently transfected with Cdh1 siRNA

    Techniques Used: Inhibition, Transfection

    TGF-β dose-dependently and time-dependently increases nuclear Cdh1 while decreasing Skp2 and Cks1 to prevent ubiquitin-mediated degradation of p27 in ECA cell lines. (A) TGF-β dose-dependently induces Cdh1 protein. ECC-1 cells, were treated
    Figure Legend Snippet: TGF-β dose-dependently and time-dependently increases nuclear Cdh1 while decreasing Skp2 and Cks1 to prevent ubiquitin-mediated degradation of p27 in ECA cell lines. (A) TGF-β dose-dependently induces Cdh1 protein. ECC-1 cells, were treated

    Techniques Used:

    The TGF-β-mediated increase in p27 and decrease in Skp2 and Cks1 do not require protein synthesis in Cdh1 wild type cells; TGF-β-induced effects are obviated in Cdh1-deficient cells and Skp2 and Cks1 half-lives are extended. (A) TGF-β
    Figure Legend Snippet: The TGF-β-mediated increase in p27 and decrease in Skp2 and Cks1 do not require protein synthesis in Cdh1 wild type cells; TGF-β-induced effects are obviated in Cdh1-deficient cells and Skp2 and Cks1 half-lives are extended. (A) TGF-β

    Techniques Used:

    31) Product Images from "Roles of Gremlin1 and Gremlin2 in Regulating Ovarian Primordial to Primary Follicle Transition"

    Article Title: Roles of Gremlin1 and Gremlin2 in Regulating Ovarian Primordial to Primary Follicle Transition

    Journal: Reproduction (Cambridge, England)

    doi: 10.1530/REP-14-0005

    GREM2 co-immunoprecipitation. A) Western blots of samples after co-IP detecting GREM2, AMH and BMP4. Lane numbers are explained in B) describing for each sample the proteins combined, the antibody used for co-immunoprecipitation and the antibody used for protein detection in the western blot.
    Figure Legend Snippet: GREM2 co-immunoprecipitation. A) Western blots of samples after co-IP detecting GREM2, AMH and BMP4. Lane numbers are explained in B) describing for each sample the proteins combined, the antibody used for co-immunoprecipitation and the antibody used for protein detection in the western blot.

    Techniques Used: Immunoprecipitation, Western Blot, Co-Immunoprecipitation Assay

    Model of GREM2 actions affecting primordial to primary follicle transition. GREM2 is secreted by granulosa cells of primordial and developing follicles. GREM2 acts to bind and inhibit the actions of BMP4 from stroma and AMH from granulosa of developing follicles.
    Figure Legend Snippet: Model of GREM2 actions affecting primordial to primary follicle transition. GREM2 is secreted by granulosa cells of primordial and developing follicles. GREM2 acts to bind and inhibit the actions of BMP4 from stroma and AMH from granulosa of developing follicles.

    Techniques Used:

    Primordial to primary follicle transition after GREM2 treatment. The follicles per ovarian cross-section were counted and categorized as either primordial or as developing (i.e. after primordial to primary follicle transition) after organ culture. Data are expressed as the proportion of developing follicles for each treatment divided by the proportion of developing follicles in the control group. N = 6–11 ovaries per treatment, from 5 experiments performed in replicate. (*) = p
    Figure Legend Snippet: Primordial to primary follicle transition after GREM2 treatment. The follicles per ovarian cross-section were counted and categorized as either primordial or as developing (i.e. after primordial to primary follicle transition) after organ culture. Data are expressed as the proportion of developing follicles for each treatment divided by the proportion of developing follicles in the control group. N = 6–11 ovaries per treatment, from 5 experiments performed in replicate. (*) = p

    Techniques Used: Organ Culture

    Ovarian immunohistochemical localization of GREM2 in 14-day old ovaries. A) GREM2 is prominently expressed in the granulosa cells of developing follicles with several layers of granulosa cells. B) Higher magnification image showing GREM2 expression in the flattened granulosa cells (arrowheads) of primordial follicles. (Inset) Higher magnification view showing of GREM2 expression in a primordial (lower) and transitional follicle. C) Negative control using non-specific IgG as a primary antibody.
    Figure Legend Snippet: Ovarian immunohistochemical localization of GREM2 in 14-day old ovaries. A) GREM2 is prominently expressed in the granulosa cells of developing follicles with several layers of granulosa cells. B) Higher magnification image showing GREM2 expression in the flattened granulosa cells (arrowheads) of primordial follicles. (Inset) Higher magnification view showing of GREM2 expression in a primordial (lower) and transitional follicle. C) Negative control using non-specific IgG as a primary antibody.

    Techniques Used: Immunohistochemistry, Expressing, Negative Control

    32) Product Images from "Roles of Gremlin1 and Gremlin2 in Regulating Ovarian Primordial to Primary Follicle Transition"

    Article Title: Roles of Gremlin1 and Gremlin2 in Regulating Ovarian Primordial to Primary Follicle Transition

    Journal: Reproduction (Cambridge, England)

    doi: 10.1530/REP-14-0005

    GREM2 co-immunoprecipitation. A) Western blots of samples after co-IP detecting GREM2, AMH and BMP4. Lane numbers are explained in B) describing for each sample the proteins combined, the antibody used for co-immunoprecipitation and the antibody used for protein detection in the western blot.
    Figure Legend Snippet: GREM2 co-immunoprecipitation. A) Western blots of samples after co-IP detecting GREM2, AMH and BMP4. Lane numbers are explained in B) describing for each sample the proteins combined, the antibody used for co-immunoprecipitation and the antibody used for protein detection in the western blot.

    Techniques Used: Immunoprecipitation, Western Blot, Co-Immunoprecipitation Assay

    Model of GREM2 actions affecting primordial to primary follicle transition. GREM2 is secreted by granulosa cells of primordial and developing follicles. GREM2 acts to bind and inhibit the actions of BMP4 from stroma and AMH from granulosa of developing follicles.
    Figure Legend Snippet: Model of GREM2 actions affecting primordial to primary follicle transition. GREM2 is secreted by granulosa cells of primordial and developing follicles. GREM2 acts to bind and inhibit the actions of BMP4 from stroma and AMH from granulosa of developing follicles.

    Techniques Used:

    Primordial to primary follicle transition after GREM2 treatment. The follicles per ovarian cross-section were counted and categorized as either primordial or as developing (i.e. after primordial to primary follicle transition) after organ culture. Data are expressed as the proportion of developing follicles for each treatment divided by the proportion of developing follicles in the control group. N = 6–11 ovaries per treatment, from 5 experiments performed in replicate. (*) = p
    Figure Legend Snippet: Primordial to primary follicle transition after GREM2 treatment. The follicles per ovarian cross-section were counted and categorized as either primordial or as developing (i.e. after primordial to primary follicle transition) after organ culture. Data are expressed as the proportion of developing follicles for each treatment divided by the proportion of developing follicles in the control group. N = 6–11 ovaries per treatment, from 5 experiments performed in replicate. (*) = p

    Techniques Used: Organ Culture

    Ovarian immunohistochemical localization of GREM2 in 14-day old ovaries. A) GREM2 is prominently expressed in the granulosa cells of developing follicles with several layers of granulosa cells. B) Higher magnification image showing GREM2 expression in the flattened granulosa cells (arrowheads) of primordial follicles. (Inset) Higher magnification view showing of GREM2 expression in a primordial (lower) and transitional follicle. C) Negative control using non-specific IgG as a primary antibody.
    Figure Legend Snippet: Ovarian immunohistochemical localization of GREM2 in 14-day old ovaries. A) GREM2 is prominently expressed in the granulosa cells of developing follicles with several layers of granulosa cells. B) Higher magnification image showing GREM2 expression in the flattened granulosa cells (arrowheads) of primordial follicles. (Inset) Higher magnification view showing of GREM2 expression in a primordial (lower) and transitional follicle. C) Negative control using non-specific IgG as a primary antibody.

    Techniques Used: Immunohistochemistry, Expressing, Negative Control

    33) Product Images from "Spinal RacGAP α-Chimaerin Is Required to Establish the Midline Barrier for Proper Corticospinal Axon Guidance"

    Article Title: Spinal RacGAP α-Chimaerin Is Required to Establish the Midline Barrier for Proper Corticospinal Axon Guidance

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3123-16.2017

    Midline EphA4(+) cells are clustered within MB holes in Chn1 KO mice. A – D , In MB holes of Chn1 KO ( Chn1 −/− ;Epha4 +/LacZ ) mouse spinal cord ( C , arrows), clusters of EphA4(+) cells were observed ( C , D ). In control ( Chn1 +/− ;Epha4 +/LacZ ) mice ( A ), a few EphA4(+) cells were observed within the ephrinB3(+) MB ( B , arrows), but these cells were not clustered and no holes were observed around these cells. Right panels in A and C are from the left panels in A and C , respectively, and indicate the location of β-gal(+) cells (magenta) and the MB (green). In this set of experiments, EphA4(+) cells (asterisks) were detected using the anti-β-gal antibody staining in the Epha4-LacZ mouse line. Cervical coronal sections (100 μm thick) at E15.5 were stained using anti-β-gal and anti-ephrinB3 antibodies and DAPI. The anti-β-gal antibody also detected blood vessels (arrowheads in B and D ). B and D are high magnification of the boxes in A and C , respectively. E – G , In Chn1 −/− ;Epha4 +/LacZ mouse spinal cord, the majority of EphA4(+) [β-gal(+)] cells located in the midline (surrounded by white dashed line) were observed in MB holes (magenta arrows), although some were also observed within the MB (cyan arrows). Cervical coronal sections (100 μm thick) at E15.5 were stained using anti-ephrinB3 and anti-β-gal antibodies and DAPI. We determined whether individual cells located in the midline were EphA4(+) and if individual midline EphA4(+) cells are located in MB holes (magenta) or within the MB (cyan) by analyzing 3D images. E – G are coronal and DV reslicing sections and midsagittal images, respectively. Horizontal yellow solid lines in E correspond to those in F and G . The position of E is indicated as a vertical yellow line in G . D, Dorsal; V, ventral; R, rostral; C, caudal. H , Quantitative analysis indicating that actual value [ratio of number of EphA4(+) cells in MB holes to number of EphA4(+) cells in DGM midline area] was significantly higher than expected value that was calculated from ratio of MB hole area to DGM midline area (MB area + MB hole area). Five sections from three Chn1 KO mice were analyzed. Paired t test, *** p
    Figure Legend Snippet: Midline EphA4(+) cells are clustered within MB holes in Chn1 KO mice. A – D , In MB holes of Chn1 KO ( Chn1 −/− ;Epha4 +/LacZ ) mouse spinal cord ( C , arrows), clusters of EphA4(+) cells were observed ( C , D ). In control ( Chn1 +/− ;Epha4 +/LacZ ) mice ( A ), a few EphA4(+) cells were observed within the ephrinB3(+) MB ( B , arrows), but these cells were not clustered and no holes were observed around these cells. Right panels in A and C are from the left panels in A and C , respectively, and indicate the location of β-gal(+) cells (magenta) and the MB (green). In this set of experiments, EphA4(+) cells (asterisks) were detected using the anti-β-gal antibody staining in the Epha4-LacZ mouse line. Cervical coronal sections (100 μm thick) at E15.5 were stained using anti-β-gal and anti-ephrinB3 antibodies and DAPI. The anti-β-gal antibody also detected blood vessels (arrowheads in B and D ). B and D are high magnification of the boxes in A and C , respectively. E – G , In Chn1 −/− ;Epha4 +/LacZ mouse spinal cord, the majority of EphA4(+) [β-gal(+)] cells located in the midline (surrounded by white dashed line) were observed in MB holes (magenta arrows), although some were also observed within the MB (cyan arrows). Cervical coronal sections (100 μm thick) at E15.5 were stained using anti-ephrinB3 and anti-β-gal antibodies and DAPI. We determined whether individual cells located in the midline were EphA4(+) and if individual midline EphA4(+) cells are located in MB holes (magenta) or within the MB (cyan) by analyzing 3D images. E – G are coronal and DV reslicing sections and midsagittal images, respectively. Horizontal yellow solid lines in E correspond to those in F and G . The position of E is indicated as a vertical yellow line in G . D, Dorsal; V, ventral; R, rostral; C, caudal. H , Quantitative analysis indicating that actual value [ratio of number of EphA4(+) cells in MB holes to number of EphA4(+) cells in DGM midline area] was significantly higher than expected value that was calculated from ratio of MB hole area to DGM midline area (MB area + MB hole area). Five sections from three Chn1 KO mice were analyzed. Paired t test, *** p

    Techniques Used: Mouse Assay, Staining

    EphA4 cells in the juxta-midline area express α2-chimaerin. In control ( Epha4 +/LacZ ) mice ( A ), EphA4 [β-gal(+)] cells in juxta-midline areas expressed α2-chimaerin ( D ), but these cells were not found in the midline ( E ) ( n = 4 sections from two mice). In EphA4 KO ( Epha4 LacZ/LacZ ) mice ( B ), ectopic mildine-EphA4 cells expressed α2-chimaerin ( F ) ( n = 4 sections from two mice). In Chn1 KO mice ( C ), there was no staining of α2-chimaerin, confirming the specificity of anti-α2-chimaerin antibody ( G ) ( n = 3 sections from two mice). Arrows in E – G indicate the position of the midline. Coronal sections of cervical cords at E15.5 were stained with anti-α2-chimaerin and anti-β-gal antibodies and DAPI. Scale bars: A – C , 40 μm; D – G , 10 μm.
    Figure Legend Snippet: EphA4 cells in the juxta-midline area express α2-chimaerin. In control ( Epha4 +/LacZ ) mice ( A ), EphA4 [β-gal(+)] cells in juxta-midline areas expressed α2-chimaerin ( D ), but these cells were not found in the midline ( E ) ( n = 4 sections from two mice). In EphA4 KO ( Epha4 LacZ/LacZ ) mice ( B ), ectopic mildine-EphA4 cells expressed α2-chimaerin ( F ) ( n = 4 sections from two mice). In Chn1 KO mice ( C ), there was no staining of α2-chimaerin, confirming the specificity of anti-α2-chimaerin antibody ( G ) ( n = 3 sections from two mice). Arrows in E – G indicate the position of the midline. Coronal sections of cervical cords at E15.5 were stained with anti-α2-chimaerin and anti-β-gal antibodies and DAPI. Scale bars: A – C , 40 μm; D – G , 10 μm.

    Techniques Used: Mouse Assay, Staining

    Ectopic midline-EphA4 cells are clustered within MB holes in Epha4 KO mice. A – D , Epha4 KO mice showed holes in the ephrinB3(+) MB ( C , arrows). In the holes, ectopic EphA4 cells [β-gal(+) cells; asterisks] were clustered ( D ). Right panels in A and C are depicted on the basis of the left panels in A and C , respectively, and indicate the location of β-gal(+) cells (magenta) and the MB (green). Cervical coronal sections (100 μm thick) in control ( Epha4 +/LacZ ) and Epha4 KO ( Epha4 LacZ/LacZ ) mice at P0 were stained with DAPI and anti-ephrinB3 and anti-β-gal antibodies ( A – D ). B and D are high magnification of boxes in A and C , respectively. E – G , In the Epha4 KO mouse spinal cord ( E , F ), the majority of EphA4 cells located in the midline (surrounded by white dashed line) were found in MB holes (magenta arrows), although some were found within the MB (cyan arrows). In a midsagittal section ( G ), distribution of EphA4 cells in MB holes (magenta) and that of EphA4 cells in MB (cyan) were plotted. Cervical coronal sections (100 μm thick) at P0 Epha4 KO mice were stained with anti-ephrinB3 and anti-β-gal antibodies and DAPI. Yellow lines in the coronal section ( E ) correspond to those in DV reslicing sections ( F ). Vertical and horizontal yellow solid lines in G indicate the position of cross-sections in E and F , respectively. H , Quantitative analysis indicated that actual value [ratio of number of EphA4 cells in MB holes to number of EphA4 cells in DGM midline area] was significantly higher than expected value that was calculated from ratio of DGM MB hole area to DGM midline area. Five sections from five Epha4 KO mice were analyzed. Paired t test, *** p
    Figure Legend Snippet: Ectopic midline-EphA4 cells are clustered within MB holes in Epha4 KO mice. A – D , Epha4 KO mice showed holes in the ephrinB3(+) MB ( C , arrows). In the holes, ectopic EphA4 cells [β-gal(+) cells; asterisks] were clustered ( D ). Right panels in A and C are depicted on the basis of the left panels in A and C , respectively, and indicate the location of β-gal(+) cells (magenta) and the MB (green). Cervical coronal sections (100 μm thick) in control ( Epha4 +/LacZ ) and Epha4 KO ( Epha4 LacZ/LacZ ) mice at P0 were stained with DAPI and anti-ephrinB3 and anti-β-gal antibodies ( A – D ). B and D are high magnification of boxes in A and C , respectively. E – G , In the Epha4 KO mouse spinal cord ( E , F ), the majority of EphA4 cells located in the midline (surrounded by white dashed line) were found in MB holes (magenta arrows), although some were found within the MB (cyan arrows). In a midsagittal section ( G ), distribution of EphA4 cells in MB holes (magenta) and that of EphA4 cells in MB (cyan) were plotted. Cervical coronal sections (100 μm thick) at P0 Epha4 KO mice were stained with anti-ephrinB3 and anti-β-gal antibodies and DAPI. Yellow lines in the coronal section ( E ) correspond to those in DV reslicing sections ( F ). Vertical and horizontal yellow solid lines in G indicate the position of cross-sections in E and F , respectively. H , Quantitative analysis indicated that actual value [ratio of number of EphA4 cells in MB holes to number of EphA4 cells in DGM midline area] was significantly higher than expected value that was calculated from ratio of DGM MB hole area to DGM midline area. Five sections from five Epha4 KO mice were analyzed. Paired t test, *** p

    Techniques Used: Mouse Assay, Staining

    Midline invasion of EphA4 cells in Epha4 KO mice ( A – Q ). A , Gfp expression vector was introduced unilaterally into the spinal cord of control ( Epha4 +/LacZ ) and Epha 4KO ( Epha4 LacZ/LacZ ) embryos at E11.5 using in utero electroporation and coronal sections of the cervical cord at E17.5 were stained using anti-β-gal and anti-ephrinB3 antibodies and DAPI. B and D are single-plane images obtained with confocal microscopy and A and C are z -projection (35-μm-thick) images. E – Q are high-magnification images of boxes in A – D . F′ – H′ , K′ – M′ , and O′ – Q′ , which are consistent with GFP(+) cells in F – H , K – M , and O – Q , respectively, indicate z -projection images of GFP signals. In the electroporated side (ipsi) of both genotypes, GFP(+);β-gal(−) (e.g., E and J ) and GFP(+);β-gal(+) cells (e.g., I and N ) were found. In the ephrinB3(+) MB of both genotypes, some GFP(+);β-gal(−) cells appeared as midline radial glia (RG; yellow arrows in A and C ; F and K ), because they extend their processes along the DV axis (arrows in F′ and K′ ) and other GFP(+);β-gal(−) cells (cyan arrows in A and C ; G , L ) were nonradial glial (non-RG) cells that extend their processes orthogonally to the DV axis (arrows in G′ and L′ ). Conversely, in the midline (mid), GFP(+);β-gal(+) cells (magenta arrows in C ; e.g., O ) were found only in Epha4 KO mice and they extended their processes orthogonally to the DV axis (arrows in O′ ). In the contralateral (contra) side, both genotypes had GFP(+);β-gal(−) cells extending their processes orthogonally to the DV axis (cyan arrowheads in A and C ; e.g., H and M ; arrows in H′ and M′ ), but only Epha4 KO mice had GFP(+);β-gal(+) cells (magenta arrowheads in C ; e.g., P ), which projected axon-like processes to the ipsilateral side (arrows in P′ and Q′ ). Asterisks in F′ – H′ , K′ – M′ , and O′ – Q′ indicate somata. The MB is outlined with green dotted lines in O′ – Q′ . Scale bars: A – D , 50 μm; E – Q , F′ – H′ , K′ – M′ , O′ – Q′ , 10 μm. R , S , Ratio of midline (Mid) GFP(+);β-gal(+) cells in total GFP(+);β-gal(+) cells in each section ( R ) and ratio of contralateral (Contra) GFP(+);β-gal(+) cells in total GFP(+);β-gal(+) cells in each section ( S ) are shown (control: n = 5 sections from 3 mice; EphA4 KO: n = 8 sections from 4 mice). Only in Epha4 KO mice were some GFP(+);β-gal(+) cells distributed in the midline and contralateral areas. Data are represented as scatter plot and mean. Mann–Whitney U test, ** p
    Figure Legend Snippet: Midline invasion of EphA4 cells in Epha4 KO mice ( A – Q ). A , Gfp expression vector was introduced unilaterally into the spinal cord of control ( Epha4 +/LacZ ) and Epha 4KO ( Epha4 LacZ/LacZ ) embryos at E11.5 using in utero electroporation and coronal sections of the cervical cord at E17.5 were stained using anti-β-gal and anti-ephrinB3 antibodies and DAPI. B and D are single-plane images obtained with confocal microscopy and A and C are z -projection (35-μm-thick) images. E – Q are high-magnification images of boxes in A – D . F′ – H′ , K′ – M′ , and O′ – Q′ , which are consistent with GFP(+) cells in F – H , K – M , and O – Q , respectively, indicate z -projection images of GFP signals. In the electroporated side (ipsi) of both genotypes, GFP(+);β-gal(−) (e.g., E and J ) and GFP(+);β-gal(+) cells (e.g., I and N ) were found. In the ephrinB3(+) MB of both genotypes, some GFP(+);β-gal(−) cells appeared as midline radial glia (RG; yellow arrows in A and C ; F and K ), because they extend their processes along the DV axis (arrows in F′ and K′ ) and other GFP(+);β-gal(−) cells (cyan arrows in A and C ; G , L ) were nonradial glial (non-RG) cells that extend their processes orthogonally to the DV axis (arrows in G′ and L′ ). Conversely, in the midline (mid), GFP(+);β-gal(+) cells (magenta arrows in C ; e.g., O ) were found only in Epha4 KO mice and they extended their processes orthogonally to the DV axis (arrows in O′ ). In the contralateral (contra) side, both genotypes had GFP(+);β-gal(−) cells extending their processes orthogonally to the DV axis (cyan arrowheads in A and C ; e.g., H and M ; arrows in H′ and M′ ), but only Epha4 KO mice had GFP(+);β-gal(+) cells (magenta arrowheads in C ; e.g., P ), which projected axon-like processes to the ipsilateral side (arrows in P′ and Q′ ). Asterisks in F′ – H′ , K′ – M′ , and O′ – Q′ indicate somata. The MB is outlined with green dotted lines in O′ – Q′ . Scale bars: A – D , 50 μm; E – Q , F′ – H′ , K′ – M′ , O′ – Q′ , 10 μm. R , S , Ratio of midline (Mid) GFP(+);β-gal(+) cells in total GFP(+);β-gal(+) cells in each section ( R ) and ratio of contralateral (Contra) GFP(+);β-gal(+) cells in total GFP(+);β-gal(+) cells in each section ( S ) are shown (control: n = 5 sections from 3 mice; EphA4 KO: n = 8 sections from 4 mice). Only in Epha4 KO mice were some GFP(+);β-gal(+) cells distributed in the midline and contralateral areas. Data are represented as scatter plot and mean. Mann–Whitney U test, ** p

    Techniques Used: Mouse Assay, Expressing, Plasmid Preparation, In Utero, Electroporation, Staining, Confocal Microscopy, MANN-WHITNEY

    34) Product Images from "Mutations in DZIP1L, which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease"

    Article Title: Mutations in DZIP1L, which encodes a ciliary transition zone protein, cause autosomal recessive polycystic kidney disease

    Journal: Nature genetics

    doi: 10.1038/ng.3871

    DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.
    Figure Legend Snippet: DZIP1L localization overlaps with basal body, centrosome and transition zone markers. (a,b) IF staining for DZIP1L (green) and acetylated-α-tubulin and γ-tubulin (both in magenta) in serum starved IMCD3 kidney cells. DZIP1L staining overlaps with the basal body in ciliated cells (a) and the centrosome in non-ciliated cells (b). (c–d) Co-localization of DZIP1L (magenta) with the distal (CEP164, green in c) and subdistal (ODF2, green in d) appendage proteins. (e–i) DZIP1L (green) tracks with the centrioles (magenta) throughout all stages of the cell cycle in IMCD3 cells. (j) TCTN1 (magenta) and DZIP1L (cyan) co-localize at the transition zone in human dermal fibroblasts. ARL13B-GFP was used to mark the ciliary membrane around the axoneme (green in j). (k) 3D-SIM superresolution microscopy on RPE-1 (human retinal pigment epithelial) cells confirms both DZIP1L (magenta) and TCTN1 (cyan) are closely associated at the transition zone. Cilium labeled with ARL13B-GFP (green). Nuclei are stained with DAPI (blue). Scale bars in a-I = 5µm; j = 1µm; k = 500 nm. DZIP1L stained with Sigma C-terminal antibody, except in panels c,d,j,k where the Abnova antibody was used.

    Techniques Used: Staining, Microscopy, Labeling

    35) Product Images from "Kinetic analysis of novel mono- and multivalent VHH-fragments and their application for molecular imaging of brain tumours"

    Article Title: Kinetic analysis of novel mono- and multivalent VHH-fragments and their application for molecular imaging of brain tumours

    Journal: British Journal of Pharmacology

    doi: 10.1111/j.1476-5381.2010.00742.x

    (A1) Immunofluorescence of anti-EGFR polyclonal antibody or (B1) V2C-EG 2 sdAb on sections of human high-grade glioma brain tumour. EGFR expression in red, DAPI staining for cell nuclei in blue and ULEX staining for brain blood vessels in green. EGFR expression only (in red) is shown in right panels (A2, B2). Scale bar: 50 µm.
    Figure Legend Snippet: (A1) Immunofluorescence of anti-EGFR polyclonal antibody or (B1) V2C-EG 2 sdAb on sections of human high-grade glioma brain tumour. EGFR expression in red, DAPI staining for cell nuclei in blue and ULEX staining for brain blood vessels in green. EGFR expression only (in red) is shown in right panels (A2, B2). Scale bar: 50 µm.

    Techniques Used: Immunofluorescence, Expressing, Staining

    EGFR and EGFRvIII expressed on U87MG cell lines were affinity labelled with [ 125 I]-EG 2 in the absence and presence of 100-fold excess unlabelled EG 2 (A, left). The resulting bands were competed for by unlabelled sdAb and corresponded to the expected molecular weight as confirmed by Western blot (A, right). IC 50 values were calculated by fitting [ 125 I]-EG 2 cell binding data to a one-site competition model using non-linear regression. For monovalent EG 2 binding to EGFR on the surface of U87MG.wtEGFR cells, the average IC 50 was 7.04 ± 2.67 × 10 −8 M (B). Similarly, the average IC 50 on U87MG.EGFRvIII cells was 5.98 ± 0.12 × 10 −8 M (C).
    Figure Legend Snippet: EGFR and EGFRvIII expressed on U87MG cell lines were affinity labelled with [ 125 I]-EG 2 in the absence and presence of 100-fold excess unlabelled EG 2 (A, left). The resulting bands were competed for by unlabelled sdAb and corresponded to the expected molecular weight as confirmed by Western blot (A, right). IC 50 values were calculated by fitting [ 125 I]-EG 2 cell binding data to a one-site competition model using non-linear regression. For monovalent EG 2 binding to EGFR on the surface of U87MG.wtEGFR cells, the average IC 50 was 7.04 ± 2.67 × 10 −8 M (B). Similarly, the average IC 50 on U87MG.EGFRvIII cells was 5.98 ± 0.12 × 10 −8 M (C).

    Techniques Used: Molecular Weight, Western Blot, Binding Assay

    36) Product Images from "The stimulatory G protein Gsα is required in melanocortin 4 receptor–expressing cells for normal energy balance, thermogenesis, and glucose metabolism"

    Article Title: The stimulatory G protein Gsα is required in melanocortin 4 receptor–expressing cells for normal energy balance, thermogenesis, and glucose metabolism

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA118.003450

    Acute responses to cold tolerance in MC4RGsKO mouse lines. Shown are rectal temperature ( Tb ) ( A ) and iBAT temperature ( B ) measured in 2-month-old male MC4RGsKO mice and controls maintained on standard diet at room temperature (RT, time 0) and hourly after being placed at 6 °C ( n = 4–5/group). Shown are rectal ( D ) and iBAT temperature ( E ), respectively, in similar experiments performed in 2-month-old male mMC4RGsKO (4–5/group). G , rectal temperature in 3–4-month-old pMC4RGsKO (11–13/group) after acute cold exposure. C, F, and G , BAT Ucp1 mRNA levels (normalized to control mice at RT) in male MC4RGsKO ( C ), mMC4RGsKO ( F ), and pMC4RGsKO ( H ) and their respective controls at room temperature (22 °C, RT) and after 6 h at 4 °C ( n = 4–6/group). Data are expressed as mean ± S.E. ( error bars ). *, p
    Figure Legend Snippet: Acute responses to cold tolerance in MC4RGsKO mouse lines. Shown are rectal temperature ( Tb ) ( A ) and iBAT temperature ( B ) measured in 2-month-old male MC4RGsKO mice and controls maintained on standard diet at room temperature (RT, time 0) and hourly after being placed at 6 °C ( n = 4–5/group). Shown are rectal ( D ) and iBAT temperature ( E ), respectively, in similar experiments performed in 2-month-old male mMC4RGsKO (4–5/group). G , rectal temperature in 3–4-month-old pMC4RGsKO (11–13/group) after acute cold exposure. C, F, and G , BAT Ucp1 mRNA levels (normalized to control mice at RT) in male MC4RGsKO ( C ), mMC4RGsKO ( F ), and pMC4RGsKO ( H ) and their respective controls at room temperature (22 °C, RT) and after 6 h at 4 °C ( n = 4–6/group). Data are expressed as mean ± S.E. ( error bars ). *, p

    Techniques Used: Mouse Assay

    Chronic cold adaptation in MC4RGsKO and mMC4RGsKO mice. A and B , rectal temperature in male MC4RGsKO (5–7-month-old) ( A ) and mMC4RGsKO (8–12-month-old) ( B ) mice and their littermate controls over 14 days while ambient temperature was slowly lowered from 22 to 6 °C and then maintained at 6 °C for the last 5 days (temperatures for each day are noted at the top ; n = 5–8 mice/group). C and D , UCP1 immunostaining of iWAT sections from MC4RGsKO ( C ) and mMC4RGsKO ( D ) male mice housed at 22 °C or at the completion of chronic cold exposure (6 °C). Scale bar , 100 μm. E and F , BAT Ucp1 mRNA expression in male MC4RGsKO ( E ) and mMC4RGsKO ( F ) mice and their controls either maintained at 22 °C (RT) or after chronic cold adaptation. Data are expressed as mean ± S.E. ( error bars ). *, p
    Figure Legend Snippet: Chronic cold adaptation in MC4RGsKO and mMC4RGsKO mice. A and B , rectal temperature in male MC4RGsKO (5–7-month-old) ( A ) and mMC4RGsKO (8–12-month-old) ( B ) mice and their littermate controls over 14 days while ambient temperature was slowly lowered from 22 to 6 °C and then maintained at 6 °C for the last 5 days (temperatures for each day are noted at the top ; n = 5–8 mice/group). C and D , UCP1 immunostaining of iWAT sections from MC4RGsKO ( C ) and mMC4RGsKO ( D ) male mice housed at 22 °C or at the completion of chronic cold exposure (6 °C). Scale bar , 100 μm. E and F , BAT Ucp1 mRNA expression in male MC4RGsKO ( E ) and mMC4RGsKO ( F ) mice and their controls either maintained at 22 °C (RT) or after chronic cold adaptation. Data are expressed as mean ± S.E. ( error bars ). *, p

    Techniques Used: Mouse Assay, Immunostaining, Expressing

    37) Product Images from "The In Ovo Delivery of CpG Oligonucleotides Protects against Infectious Bronchitis with the Recruitment of Immune Cells into the Respiratory Tract of Chickens"

    Article Title: The In Ovo Delivery of CpG Oligonucleotides Protects against Infectious Bronchitis with the Recruitment of Immune Cells into the Respiratory Tract of Chickens

    Journal: Viruses

    doi: 10.3390/v10110635

    In ovo delivery of CpG ODNs is capable of recruiting key cells of the innate and adaptive arms of the immune system responsible for enhanced immune responses in the respiratory tract. The quantitative data following immunofluorescent assays done for the trachea ( a ) macrophages, ( b ) cluster of differentiation (CD)4+ T cells, and (c) CD8α+ T cells are given. The quantitative data following the immunofluorescent assays done for lung ( d ) macrophages, ( e ) CD4+ T cells, and ( f ) CD8α+ T cells are given. One-way ANOVA followed by the Students–Newman–Keuls post hoc test were used to identify the group differences. The differences were considered significant at * = significant at  p  ≤ 0.05, ** = significant at  p  ≤ 0.01 *** = significant at  p  ≤ 0.001. The results represent the pooled data of two independent experiments.
    Figure Legend Snippet: In ovo delivery of CpG ODNs is capable of recruiting key cells of the innate and adaptive arms of the immune system responsible for enhanced immune responses in the respiratory tract. The quantitative data following immunofluorescent assays done for the trachea ( a ) macrophages, ( b ) cluster of differentiation (CD)4+ T cells, and (c) CD8α+ T cells are given. The quantitative data following the immunofluorescent assays done for lung ( d ) macrophages, ( e ) CD4+ T cells, and ( f ) CD8α+ T cells are given. One-way ANOVA followed by the Students–Newman–Keuls post hoc test were used to identify the group differences. The differences were considered significant at * = significant at p ≤ 0.05, ** = significant at p ≤ 0.01 *** = significant at p ≤ 0.001. The results represent the pooled data of two independent experiments.

    Techniques Used: In Ovo

    38) Product Images from "Pretangle pathology within cholinergic nucleus basalis neurons coincides with neurotrophic and neurotransmitter receptor gene dysregulation during the progression of Alzheimer’s disease"

    Article Title: Pretangle pathology within cholinergic nucleus basalis neurons coincides with neurotrophic and neurotransmitter receptor gene dysregulation during the progression of Alzheimer’s disease

    Journal: Neurobiology of disease

    doi: 10.1016/j.nbd.2018.05.021

    The expression of neurotrophin receptor and select downstream signaling molecule gene transcripts is equivalent in pS422+ nbM neurons during the progression of AD. Color-coded heatmap of the relative expression profiles for select transcripts in pS422+ nbM neurons aspirated from NCI, MCI, and AD cases (red to green = increasing mRNA levels). Quantitative analysis revealed no statistical differences in the expression levels of the transcripts examined in pS422+ nbM neurons derived from MCI or AD compared to NCI. This observation suggests that the pathological state of the neuron, not disease status, may drive changes in gene expression. Therefore, in the present analysis, we compared mRNA levels in individual pS422+, pS422+/TauC3+, and TauC3+ nbM neurons independent of clinical diagnosis. Abbreviations: Nrtk1, Nrtk2 , and Nrtk3 , neurotrophin tyrosine kinase receptor type 1 (TrkA), 2 (TrkB), 3 (TrkC); ECD, extracellular domain; TK, intracellular tyrosine kinase domain; Ngfr , nerve growth factor receptor (p75 NTR ); Mapk1 , mitogen-activated protein kinase 1 (extracellular signal-regulated kinase 2); Mapk3 , mitogen-activated protein kinase 3 (extracellular signal-regulated kinase 1); Creb1 , cAMP response element binding protein; Akt1 , Akt serine/threonine kinase 1 (protein kinase B); Prkca, Prkce, Prkci , protein kinase C alpha, epsilon, iota.
    Figure Legend Snippet: The expression of neurotrophin receptor and select downstream signaling molecule gene transcripts is equivalent in pS422+ nbM neurons during the progression of AD. Color-coded heatmap of the relative expression profiles for select transcripts in pS422+ nbM neurons aspirated from NCI, MCI, and AD cases (red to green = increasing mRNA levels). Quantitative analysis revealed no statistical differences in the expression levels of the transcripts examined in pS422+ nbM neurons derived from MCI or AD compared to NCI. This observation suggests that the pathological state of the neuron, not disease status, may drive changes in gene expression. Therefore, in the present analysis, we compared mRNA levels in individual pS422+, pS422+/TauC3+, and TauC3+ nbM neurons independent of clinical diagnosis. Abbreviations: Nrtk1, Nrtk2 , and Nrtk3 , neurotrophin tyrosine kinase receptor type 1 (TrkA), 2 (TrkB), 3 (TrkC); ECD, extracellular domain; TK, intracellular tyrosine kinase domain; Ngfr , nerve growth factor receptor (p75 NTR ); Mapk1 , mitogen-activated protein kinase 1 (extracellular signal-regulated kinase 2); Mapk3 , mitogen-activated protein kinase 3 (extracellular signal-regulated kinase 1); Creb1 , cAMP response element binding protein; Akt1 , Akt serine/threonine kinase 1 (protein kinase B); Prkca, Prkce, Prkci , protein kinase C alpha, epsilon, iota.

    Techniques Used: Expressing, Derivative Assay, Binding Assay

    Phenotypic characterization of NFT evolution with pS422 and TauC3 immunoreactivity in nbM neurons. (A–G) Tissue sections from the nbM of a representative AD case. (A) Cholinergic neurons in the nbM can be identified phenotypically by expression of the pan-neurotrophin (p75 NTR ) receptor. A tissue section from a consecutive series was immunostained with p75 NTR (brown) and pS422 (blue) to confirm the location of the cholinergic nbM. (B) Low magnification view of the nbM subfield immunostained with pS422 (brown) and TauC3 (blue). (C) High magnification of pS422 and TauC3 pathology in boxed area from (B). A Nissl counterstain was used to identify nbM neurons lacking tau pathology (*). (D-G) Confocal microscopy was used to confirm the presence of three discrete populations of nbM neurons. (D) Low magnification view of nbM subfield. (E–G) High magnification of boxed area from (D) identify pS422 (E), TauC3 (F), and overlay (G). Single arrowhead indicates a pS422+ nbM neuron, double arrowheads indicate a TauC3+ nbM neuron, and arrow indicates a pS422+/TauC3+ nbM neuron (colocalization appears yellow). Scale bar in A, 100 μm for A–B; scale bar in C, 50 μm for C; scale bar in D, 100 μm for D; scale bar in G, 50 μm for E–G.
    Figure Legend Snippet: Phenotypic characterization of NFT evolution with pS422 and TauC3 immunoreactivity in nbM neurons. (A–G) Tissue sections from the nbM of a representative AD case. (A) Cholinergic neurons in the nbM can be identified phenotypically by expression of the pan-neurotrophin (p75 NTR ) receptor. A tissue section from a consecutive series was immunostained with p75 NTR (brown) and pS422 (blue) to confirm the location of the cholinergic nbM. (B) Low magnification view of the nbM subfield immunostained with pS422 (brown) and TauC3 (blue). (C) High magnification of pS422 and TauC3 pathology in boxed area from (B). A Nissl counterstain was used to identify nbM neurons lacking tau pathology (*). (D-G) Confocal microscopy was used to confirm the presence of three discrete populations of nbM neurons. (D) Low magnification view of nbM subfield. (E–G) High magnification of boxed area from (D) identify pS422 (E), TauC3 (F), and overlay (G). Single arrowhead indicates a pS422+ nbM neuron, double arrowheads indicate a TauC3+ nbM neuron, and arrow indicates a pS422+/TauC3+ nbM neuron (colocalization appears yellow). Scale bar in A, 100 μm for A–B; scale bar in C, 50 μm for C; scale bar in D, 100 μm for D; scale bar in G, 50 μm for E–G.

    Techniques Used: Expressing, Confocal Microscopy

    Neurotrophin receptor and select downstream signaling molecule mRNAs are dysregulated during the progression of NFT maturation. Heatmap of relative mRNA expression levels of neurotrophin receptors and downstream signaling molecules in pS422+, pS422+/TauC3+, and TauC3+ nbM neurons compared to unlabeled nbM neurons (red to green = increasing mRNA levels). Quantitative analysis revealed downregulated expression of Nrtk1-3 transcripts as well as Akt1 and Prkce in pS422+ nbM neurons as compared to unlabeled control neurons. Appearance of the late stage neoepitope TauC3 was associated with downregulation of the Ngfr transcript and upregulation of the Prkca transcript. a, unlabeled > pS422, p
    Figure Legend Snippet: Neurotrophin receptor and select downstream signaling molecule mRNAs are dysregulated during the progression of NFT maturation. Heatmap of relative mRNA expression levels of neurotrophin receptors and downstream signaling molecules in pS422+, pS422+/TauC3+, and TauC3+ nbM neurons compared to unlabeled nbM neurons (red to green = increasing mRNA levels). Quantitative analysis revealed downregulated expression of Nrtk1-3 transcripts as well as Akt1 and Prkce in pS422+ nbM neurons as compared to unlabeled control neurons. Appearance of the late stage neoepitope TauC3 was associated with downregulation of the Ngfr transcript and upregulation of the Prkca transcript. a, unlabeled > pS422, p

    Techniques Used: Expressing

    Select cholinergic markers are dysregulated during the development of NFTs in nbM neurons. Heatmap of relative expression levels of cholinergic neuronal markers in pS422+, pS422+/TauC3+, and TauC3+ NB neurons compared to unlabeled control neurons (red to green = increasing mRNA levels). Quantitative analysis revealed upregulation of the Chrna7 transcript and downregulation of the Chrnb2 transcript following the appearance of the TauC3 epitope. Abbreviations: Chat choline acetyltransferase; Slcl8a3 , vesicular acetylcholine transporter; Ache , acetylcholinesterase; Bche , butyrylcholinesterase; Chrm1, Chrm2 , cholinergic receptor, muscarinic 1, 2; Chrna7, Chrna4, Chrnb2 , cholinergic receptor, nicotinic, alpha polypeptide 7, alpha polypeptide 4, beta polypeptide 2. a, pS422
    Figure Legend Snippet: Select cholinergic markers are dysregulated during the development of NFTs in nbM neurons. Heatmap of relative expression levels of cholinergic neuronal markers in pS422+, pS422+/TauC3+, and TauC3+ NB neurons compared to unlabeled control neurons (red to green = increasing mRNA levels). Quantitative analysis revealed upregulation of the Chrna7 transcript and downregulation of the Chrnb2 transcript following the appearance of the TauC3 epitope. Abbreviations: Chat choline acetyltransferase; Slcl8a3 , vesicular acetylcholine transporter; Ache , acetylcholinesterase; Bche , butyrylcholinesterase; Chrm1, Chrm2 , cholinergic receptor, muscarinic 1, 2; Chrna7, Chrna4, Chrnb2 , cholinergic receptor, nicotinic, alpha polypeptide 7, alpha polypeptide 4, beta polypeptide 2. a, pS422

    Techniques Used: Expressing

    39) Product Images from "BTBR ob/ob mouse model of type 2 diabetes exhibits early loss of retinal function and retinal inflammation followed by late vascular changes"

    Article Title: BTBR ob/ob mouse model of type 2 diabetes exhibits early loss of retinal function and retinal inflammation followed by late vascular changes

    Journal: Diabetologia

    doi: 10.1007/s00125-018-4696-x

    ( a ) GFAP is found along the GCL, and Müller cell processes are seen to project towards the plexiform layer. Representative images of GFAP expression (red) in transverse sections of eyes from 6-week-old  ob − / ob −  and  ob + / ob +  mice reveal elevated GFAP expression in  ob − / ob −  mice along the GCL (white arrowheads) and OPL (yellow arrowheads), suggesting gliosis. Scale bar, 100 μm. ( b ) In the retinal whole-mounts, there were more GFAP-labelled Müller cells (green) in the primary plexus layer in  ob − / ob −  than  ob + / ob +  mice. Gliosis by upregulated Müller cells reflects inflammation of the nervous system. Scale bar, 50 μm. ( c ) In the same layer of retinal whole-mounts, the IBA-1-labelled macrophages (green) were more abundant, amoeboid and activated in  ob − / ob −  mice. Scale bars, 150 μm. ( d ) Graph of the number of Rho-Con A-labelled leucocytes in the main retinal blood vessels in each eye, in 6- and 20-week-old  ob + / ob +  and  ob − / ob −  mice. In both age groups, there were more leucocytes in the eyes of  ob − / ob −  than  ob + / ob +  mice. This was significant at both 6 weeks ( n  = 6 eyes) and 20 weeks ( n  = 7 eyes); * p
    Figure Legend Snippet: ( a ) GFAP is found along the GCL, and Müller cell processes are seen to project towards the plexiform layer. Representative images of GFAP expression (red) in transverse sections of eyes from 6-week-old ob − / ob − and ob + / ob + mice reveal elevated GFAP expression in ob − / ob − mice along the GCL (white arrowheads) and OPL (yellow arrowheads), suggesting gliosis. Scale bar, 100 μm. ( b ) In the retinal whole-mounts, there were more GFAP-labelled Müller cells (green) in the primary plexus layer in ob − / ob − than ob + / ob + mice. Gliosis by upregulated Müller cells reflects inflammation of the nervous system. Scale bar, 50 μm. ( c ) In the same layer of retinal whole-mounts, the IBA-1-labelled macrophages (green) were more abundant, amoeboid and activated in ob − / ob − mice. Scale bars, 150 μm. ( d ) Graph of the number of Rho-Con A-labelled leucocytes in the main retinal blood vessels in each eye, in 6- and 20-week-old ob + / ob + and ob − / ob − mice. In both age groups, there were more leucocytes in the eyes of ob − / ob − than ob + / ob + mice. This was significant at both 6 weeks ( n  = 6 eyes) and 20 weeks ( n  = 7 eyes); * p

    Techniques Used: Expressing, Mouse Assay

    40) Product Images from "Dlx1 and Rgs5 in the Ductus Arteriosus: Vessel-Specific Genes Identified by Transcriptional Profiling of Laser-Capture Microdissected Endothelial and Smooth Muscle Cells"

    Article Title: Dlx1 and Rgs5 in the Ductus Arteriosus: Vessel-Specific Genes Identified by Transcriptional Profiling of Laser-Capture Microdissected Endothelial and Smooth Muscle Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086892

    Gene expression of Rgs5 (a), Dlx1 (b), Tfap2B (c) and Pcp4 (d) by microarray. Expression levels are expressed as fluorescent signal intensity measured on the array after normalization. The dots represent individual samples. Horizontal bars represent the means. The same colors are used in a–d. Red = ECs from the aorta at day 18 (AO EC 18), yellow = ECs from the aorta at day 21 (AO EC 21), light green = SMCs from the aorta at day 18 (AO SMC 18), dark green = SMCs from the aorta at day 21 (AO SMC 21), turquoise = ECs from the DA at day 18 (DA EC 18), blue = ECs from the DA at day 21 (DA EC 21), purple = SMCs from the DA at day 18 (DA SMC 18), pink = SMCs from the DA at day 21).
    Figure Legend Snippet: Gene expression of Rgs5 (a), Dlx1 (b), Tfap2B (c) and Pcp4 (d) by microarray. Expression levels are expressed as fluorescent signal intensity measured on the array after normalization. The dots represent individual samples. Horizontal bars represent the means. The same colors are used in a–d. Red = ECs from the aorta at day 18 (AO EC 18), yellow = ECs from the aorta at day 21 (AO EC 21), light green = SMCs from the aorta at day 18 (AO SMC 18), dark green = SMCs from the aorta at day 21 (AO SMC 21), turquoise = ECs from the DA at day 18 (DA EC 18), blue = ECs from the DA at day 21 (DA EC 21), purple = SMCs from the DA at day 18 (DA SMC 18), pink = SMCs from the DA at day 21).

    Techniques Used: Expressing, Microarray

    Visual representation of microarray results. A. Spectral map bioplot. The first two principal components (PC) of the weighted spectral map analysis (SPM) of normalized microarray data are plotted. The samples are depicted in coloured squares with numbers. The colours are explained in the figure. AO SMC = smooth muscle cells from the descending aorta, DA EC = endothelial cells from the ductus arteriosus, DA SMC = smooth muscle cells from the ductus arteriosus. 18 d = day 18 of gestation, 21 d = day 21 of gestation. Distances between the squares are a measure for the similarity between samples. Genes that do not contribute to the differences are indicated as dots in the cloud around the centroid (represented by the cross). The ten most significantly contributing genes are annotated by their gene symbol. The first PC (PC1) explains 29% of the variance in the dataset and discriminates samples from day 18 (n = 24) from those of day 21 (n = 24). The second PC (PC2) explains 8% of the variance and discriminates between ECs and SMCs. B. Histogram showing the most significant differentially expressed genes between DA and aorta. The annotation of the genes is on the x-axis. The adjusted p-values are on the y-axis. Red bars represent genes that are enriched in the aorta. Green bars represent genes that are upregulated in the DA. C. Volcano plot. The volcano plot constructed with LIMMA analysis summarizes the fold changes between the two types of the samples ( i.e. , DA versus aorta) and the log10 transformed p-values. The negative log10 transformed p-values (y-axis) are plotted against the log ratios between the samples (log 2 fold change). For our study we selected 4 genes. The position in the upper left ( Rgs5, Tfap2B, Dlx1 ) is the result of a high ratio of differential expression.
    Figure Legend Snippet: Visual representation of microarray results. A. Spectral map bioplot. The first two principal components (PC) of the weighted spectral map analysis (SPM) of normalized microarray data are plotted. The samples are depicted in coloured squares with numbers. The colours are explained in the figure. AO SMC = smooth muscle cells from the descending aorta, DA EC = endothelial cells from the ductus arteriosus, DA SMC = smooth muscle cells from the ductus arteriosus. 18 d = day 18 of gestation, 21 d = day 21 of gestation. Distances between the squares are a measure for the similarity between samples. Genes that do not contribute to the differences are indicated as dots in the cloud around the centroid (represented by the cross). The ten most significantly contributing genes are annotated by their gene symbol. The first PC (PC1) explains 29% of the variance in the dataset and discriminates samples from day 18 (n = 24) from those of day 21 (n = 24). The second PC (PC2) explains 8% of the variance and discriminates between ECs and SMCs. B. Histogram showing the most significant differentially expressed genes between DA and aorta. The annotation of the genes is on the x-axis. The adjusted p-values are on the y-axis. Red bars represent genes that are enriched in the aorta. Green bars represent genes that are upregulated in the DA. C. Volcano plot. The volcano plot constructed with LIMMA analysis summarizes the fold changes between the two types of the samples ( i.e. , DA versus aorta) and the log10 transformed p-values. The negative log10 transformed p-values (y-axis) are plotted against the log ratios between the samples (log 2 fold change). For our study we selected 4 genes. The position in the upper left ( Rgs5, Tfap2B, Dlx1 ) is the result of a high ratio of differential expression.

    Techniques Used: Microarray, Construct, Transformation Assay, Expressing

    Gene expression of Rgs5 (a), Dlx1 (b), Tfap2B (c) and Pcp4 (d) by rtqPCR. The same mRNA preparations were used for microarray (shown in figure 4 ) and rtqPCR. Relative expression levels are shown for each sample. Horizontal bars depict the means. The levels are peptidylprolylisomerase B ( Ppib ) normalized. Red symbols represent endothelial cells (EC) and green symbols represent smooth muscle cells (SMC). AO = aorta, DA = ductus arteriosus. 18 = day 18, 21 = day 21.
    Figure Legend Snippet: Gene expression of Rgs5 (a), Dlx1 (b), Tfap2B (c) and Pcp4 (d) by rtqPCR. The same mRNA preparations were used for microarray (shown in figure 4 ) and rtqPCR. Relative expression levels are shown for each sample. Horizontal bars depict the means. The levels are peptidylprolylisomerase B ( Ppib ) normalized. Red symbols represent endothelial cells (EC) and green symbols represent smooth muscle cells (SMC). AO = aorta, DA = ductus arteriosus. 18 = day 18, 21 = day 21.

    Techniques Used: Expressing, Microarray

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    Avidin-Biotin Assay:

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    Incubation:

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    Article Title: High Mobility Group A2 (HMGA2) Promotes EMT via MAPK Pathway in Prostate Cancer
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    Article Snippet: Phaseolus vulgaris leucoagglutinin (PHA-L), goat anti-PHA-L, and FITC-conjugated streptavidin were purchased from Vector Laboratories (Burlingame, CA, USA).

    Staining:

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    Hybridization:

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    Software:

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    Vector Laboratories vectashield mounting medium
    The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into <t>Vectashield</t> mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.
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    The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Article Snippet: After multiple washes, the coverslips were mounted into Vectashield mounting medium (with DAPI for anti-mCherry antibody staining).

    Techniques: Marker, Transfection, Mutagenesis, Variant Assay, Microscopy

    Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Article Snippet: After multiple washes, the coverslips were mounted into Vectashield mounting medium (with DAPI for anti-mCherry antibody staining).

    Techniques: Transfection, Expressing, Plasmid Preparation, Incubation, Microscopy

    Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Article Snippet: After multiple washes, the coverslips were mounted into Vectashield mounting medium (with DAPI for anti-mCherry antibody staining).

    Techniques: Transfection, Marker, Microscopy

    STORM imaging of microtubules (see section 2.2 for more details) in Vectashield. (A): Widefield image (B): Single frame, (C1): Reconstructed STORM image, with blow-up on the ROI in (C2). scale-bar = 5 μ m for (A),(B),(C1) and 1 μ m for C2.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: STORM imaging of microtubules (see section 2.2 for more details) in Vectashield. (A): Widefield image (B): Single frame, (C1): Reconstructed STORM image, with blow-up on the ROI in (C2). scale-bar = 5 μ m for (A),(B),(C1) and 1 μ m for C2.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Imaging

    STORM imaging of Alexa-647 stained microtubules in a Vectashield/TRIS-Glycerol mixture: (A) 50% Vectashield and (B) 25% Vectashield. The different panels represent: (1) STORM image reconstructed from 15.000 frames, scale-bar = 500 nm (2) photon count distribution per frame and per molecule, averaged over three data-sets, and (3) standard deviation of multiple localizations giving a measure of the frame localization precision.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: STORM imaging of Alexa-647 stained microtubules in a Vectashield/TRIS-Glycerol mixture: (A) 50% Vectashield and (B) 25% Vectashield. The different panels represent: (1) STORM image reconstructed from 15.000 frames, scale-bar = 500 nm (2) photon count distribution per frame and per molecule, averaged over three data-sets, and (3) standard deviation of multiple localizations giving a measure of the frame localization precision.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Imaging, Staining, Standard Deviation

    (A) Absorption spectrum of Vectashield, as well as normalized emission spectra measured at 3 different wavelengths: 400 nm (B), 560 nm (C) and 630 nm (D) with normalization factor indicated in the top right corner.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: (A) Absorption spectrum of Vectashield, as well as normalized emission spectra measured at 3 different wavelengths: 400 nm (B), 560 nm (C) and 630 nm (D) with normalization factor indicated in the top right corner.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques:

    Statistics on STORM imaging performed in 25% Vectashield - 75% TRIS-Glycerol in which were added 1% NPG (w/v) (A), 20 mM DABCO (B), and 10 mM Lipoic Acid (C). The different panels represent: (1) photon count distribution per frame and per molecule, averaged over three datasets, (2) standard deviation of multiple localizations giving a measure of the frame localization precision, and (3) Density of molecules as a function of number of recorded frames, averaged over three measurements, with error bars indicating the standard deviation.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: Statistics on STORM imaging performed in 25% Vectashield - 75% TRIS-Glycerol in which were added 1% NPG (w/v) (A), 20 mM DABCO (B), and 10 mM Lipoic Acid (C). The different panels represent: (1) photon count distribution per frame and per molecule, averaged over three datasets, (2) standard deviation of multiple localizations giving a measure of the frame localization precision, and (3) Density of molecules as a function of number of recorded frames, averaged over three measurements, with error bars indicating the standard deviation.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Imaging, Standard Deviation

    Quantifying the quality of Vectashield as a STORM buffer for Alexa-647: (A) photon count distribution per frame (blue) and per molecule (red), obtained by grouping consecutive frame localizations and (B) standard deviation of multiple localizations (see section 2.3 for grouping details), with mean values displayed in the top right corner (C) Density of molecules as a function of number of recorded frames, averaged over three measurements. The error bar indicates the standard deviation. (D) STORM image of microtubule, on which the hollowness of the structure can be resolved, as quantified in the profile taken over the 200 nm yellow-boxed region with ≈ 35 nm between the two peaks, consistent with a 25 mm structure broadened by the antibodies.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: Quantifying the quality of Vectashield as a STORM buffer for Alexa-647: (A) photon count distribution per frame (blue) and per molecule (red), obtained by grouping consecutive frame localizations and (B) standard deviation of multiple localizations (see section 2.3 for grouping details), with mean values displayed in the top right corner (C) Density of molecules as a function of number of recorded frames, averaged over three measurements. The error bar indicates the standard deviation. (D) STORM image of microtubule, on which the hollowness of the structure can be resolved, as quantified in the profile taken over the 200 nm yellow-boxed region with ≈ 35 nm between the two peaks, consistent with a 25 mm structure broadened by the antibodies.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Standard Deviation

    STORM images obtained with the other working dyes (A) Alexa-555 in 20% Vectashield-80% TRIS-Glycerol (B) Cy-5 (C) CF-647 (D) Alexa-700, all in pure Vec-tashield. scale-bar = 5 μ m.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: STORM images obtained with the other working dyes (A) Alexa-555 in 20% Vectashield-80% TRIS-Glycerol (B) Cy-5 (C) CF-647 (D) Alexa-700, all in pure Vec-tashield. scale-bar = 5 μ m.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques:

    (A) STORM image of CEP-152 stained with Cy3 using a buffer 40% Vectashield + 1% NPG + 20 mM DABCO, which improves the quality of Cy3 blinking. Scale-bar = 500 nm (B) Radial intensity distribution measured from the yellow ROI defined in (A), and Lorentzian fit showing a peak at r = 143 nm.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: (A) STORM image of CEP-152 stained with Cy3 using a buffer 40% Vectashield + 1% NPG + 20 mM DABCO, which improves the quality of Cy3 blinking. Scale-bar = 500 nm (B) Radial intensity distribution measured from the yellow ROI defined in (A), and Lorentzian fit showing a peak at r = 143 nm.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Staining

    (A) Index matching with Vectashield: Optical index as a function of Vectashield concentration starting from PBS (red) or TDE (blue), and imaging performed at n = 1.5 (adapted to oil objectives) and n=1.4 (adapted to glycerol objectives) (B–D) STORM imaging of microtubules immunostained with Alexa-647 for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively, scale-bar = 500 nm (C–E): photon count distribution per frame and per molecule, averaged over three datasets for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively.

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: (A) Index matching with Vectashield: Optical index as a function of Vectashield concentration starting from PBS (red) or TDE (blue), and imaging performed at n = 1.5 (adapted to oil objectives) and n=1.4 (adapted to glycerol objectives) (B–D) STORM imaging of microtubules immunostained with Alexa-647 for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively, scale-bar = 500 nm (C–E): photon count distribution per frame and per molecule, averaged over three datasets for the 25% Vectashield-75% TDE buffer and 50% Vectashield - 50% PBS buffer respectively.

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Concentration Assay, Imaging

    3D STORM of Alexa-647-labeled microtubules in Vectashield: (A) Imaging performed in 25% Vectashield-75 % TRIS-Glycerol, scale-bar = 5 μ m. (B1 2): axial profile taken from the two regions delimited in A (yellow for (B1), showing a single microtubule; red for (B2) showing two well-resolved microtubules crossing at a distance of ≈ 160 nm).

    Journal: Biomedical Optics Express

    Article Title: Simple buffers for 3D STORM microscopy

    doi: 10.1364/BOE.4.000885

    Figure Lengend Snippet: 3D STORM of Alexa-647-labeled microtubules in Vectashield: (A) Imaging performed in 25% Vectashield-75 % TRIS-Glycerol, scale-bar = 5 μ m. (B1 2): axial profile taken from the two regions delimited in A (yellow for (B1), showing a single microtubule; red for (B2) showing two well-resolved microtubules crossing at a distance of ≈ 160 nm).

    Article Snippet: Imaging was performed by placing the 25 mm coverslip into a holder, then pipetting 30 μ L of Vectashield (Vectorlab, H-1000) on top of it and adding a clean 18 mm coverslip to spread the Vectashield evenly.

    Techniques: Labeling, Imaging

    Wls loss in motoneuron does not enhance synapse number in single muscle fiber. ( A ) Representative images of individual gastrocnemius muscle fibers of 2-month-old control and HB9-Wls -/- mice. To visualize AChR clusters on single muscle fibers, gastrocnemius was fixed in 4% PFA and stained with R-BTX (Red) and DAPI (Blue) to show AChR clusters and myonuclei. Muscles were washed three times in PBS and teased into single fibers and mounted in Vectashield mounting medium. We counted the number of AChR cluster in each single muscle fiber and defined that synapse elimination was impaired when more than one AChR cluster was found after P15. ( B ) Quantitative analysis of NMJ number per muscle fiber. NMJ number per muscle fiber was comparable between control and HB9-Wls -/- mice at P0, P7, P15, and P60. n = 3 mice per group. Unpaired t-test, p > 0.05.

    Journal: eLife

    Article Title: Motoneuron Wnts regulate neuromuscular junction development

    doi: 10.7554/eLife.34625

    Figure Lengend Snippet: Wls loss in motoneuron does not enhance synapse number in single muscle fiber. ( A ) Representative images of individual gastrocnemius muscle fibers of 2-month-old control and HB9-Wls -/- mice. To visualize AChR clusters on single muscle fibers, gastrocnemius was fixed in 4% PFA and stained with R-BTX (Red) and DAPI (Blue) to show AChR clusters and myonuclei. Muscles were washed three times in PBS and teased into single fibers and mounted in Vectashield mounting medium. We counted the number of AChR cluster in each single muscle fiber and defined that synapse elimination was impaired when more than one AChR cluster was found after P15. ( B ) Quantitative analysis of NMJ number per muscle fiber. NMJ number per muscle fiber was comparable between control and HB9-Wls -/- mice at P0, P7, P15, and P60. n = 3 mice per group. Unpaired t-test, p > 0.05.

    Article Snippet: Muscles were washed in PBS and teased into single fibers and mounted with Vectashield mounting medium.

    Techniques: Mouse Assay, Staining

    The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: The colocalization of mCherryTM and cis/medial Golgi marker GOLM1. COS-1 cells were transfected with pSARMXmCherryTM WT or mutant variant. 48 h later, they were fixed with 4% formaldehyde and permeabilized with Tween 20. All samples were immunostained with primary rabbit anti-GOLM1 antibody and then with secondary antibody against rabbit IgG conjugated with Alexa Fluor TM Plus 488 and mounted into Vectashield mounting medium with DAPI. Samples were imaged with spinning disk confocal microscope (Andor). Magnification 600×; scale bar 20 µm.

    Article Snippet: The coverslips were mounted in Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA).

    Techniques: Marker, Transfection, Mutagenesis, Variant Assay, Microscopy

    Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: Plasma membrane distribution of M-PMV Env. The COS-1 cells were either transfected with plasmids encoding M-PMV genomic DNA with: WT Env (pSARMX-WT; panel A ), I18A Env (pSARMX-I18A; panel B ), or Y22A Env (pSARMX-Y22A; panel C ) or an M-PMV Env expression vector (pTMT-WT; panel D ). 48 h post-transfection cells were incubated with goat anti-M-PMV antibody on ice for 25 min to bind surface exposed M-PMV Env. Formaldehyde (4%) fixed cells were immunostained for CA protein with rabbit anti-CA antibody (except for the cells transfected with pTMT-WT,). Env was visualized using secondary anti-goat IgG antibody conjugated with Alexa Fluor ® 350. Samples were mounted in Vectashield mounting media and imaged on an Olympus cell^R microscope. The original blue color of the AF350 signal was changed to cyan for increased contrast. Magnification 600×; scale bars 20 µm.

    Article Snippet: The coverslips were mounted in Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA).

    Techniques: Transfection, Expressing, Plasmid Preparation, Incubation, Microscopy

    Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Journal: Viruses

    Article Title: Mason-Pfizer Monkey Virus Envelope Glycoprotein Cycling and Its Vesicular Co-Transport with Immature Particles

    doi: 10.3390/v10100575

    Figure Lengend Snippet: Rab-markers based identification of intracellular vesicles carrying mCherryTM protein variants. The COS-1 cells co-transfected with a combination of pSARMXmCherryTM and enhanced green fluorescent protein (EGFP)-tagged endosomal marker coding plasmids were fixed with 4% formaldehyde 24 h post-transfection. Upon mounting in Vectashield media the samples were imaged using an Olympus cell^R microscope. White arrowheads indicate colocalization of the mCherryTM signal and the corresponding marker. Magnification 600×; scale bars 20 µm.

    Article Snippet: The coverslips were mounted in Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA).

    Techniques: Transfection, Marker, Microscopy