fascin 1  (Vector Laboratories)


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    Name:
    Biotinylated Rabbit Anti Rat IgG Antibody
    Description:
    Biotinylated Rabbit Anti Rat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Rat IgG H L is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    Catalog Number:
    ba-4000
    Price:
    None
    Host:
    Rabbit
    Size:
    1 5 mg
    Category:
    Antibodies
    Reactivity:
    Rat
    Buy from Supplier


    Structured Review

    Vector Laboratories fascin 1
    Biotinylated Rabbit Anti Rat IgG Antibody
    Biotinylated Rabbit Anti Rat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Rat IgG H L is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    https://www.bioz.com/result/fascin 1/product/Vector Laboratories
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    Price from $9.99 to $1999.99
    fascin 1 - by Bioz Stars, 2020-09
    89/100 stars

    Images

    1) Product Images from "Fascin-1, Ezrin and Paxillin Contribute to the Malignant Progression and Are Predictors of Clinical Prognosis in Laryngeal Squamous Cell Carcinoma"

    Article Title: Fascin-1, Ezrin and Paxillin Contribute to the Malignant Progression and Are Predictors of Clinical Prognosis in Laryngeal Squamous Cell Carcinoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0050710

    Quantitative RT-PCR analysis of relative mRNA level of fascin-1 (A), ezrin (C) and paxillin (E) in laryngeal squamous cell carcinomas (LSCC) and adjacent normal margin (ANM) tissue by sample pairs. Levels were relative to that of 18s RNA as an internal control. Data are mean±SD from experiments performed in triplicate.
    Figure Legend Snippet: Quantitative RT-PCR analysis of relative mRNA level of fascin-1 (A), ezrin (C) and paxillin (E) in laryngeal squamous cell carcinomas (LSCC) and adjacent normal margin (ANM) tissue by sample pairs. Levels were relative to that of 18s RNA as an internal control. Data are mean±SD from experiments performed in triplicate.

    Techniques Used: Quantitative RT-PCR

    Survival curves by Kaplan-Meier analysis (Log-rank test). LSCC patients with negative or low expression of (A) fascin-1, (B) ezrin or (C) paxillin survived significantly longer than those with high expression.
    Figure Legend Snippet: Survival curves by Kaplan-Meier analysis (Log-rank test). LSCC patients with negative or low expression of (A) fascin-1, (B) ezrin or (C) paxillin survived significantly longer than those with high expression.

    Techniques Used: Expressing

    Representative immunohistochemistry staining of fascin-1 expression in LSCC. Positive immunostaining of fascin-1 is in cytoplasm of tumor cells (pink arrows) and in endothelial cells of microvessels (blue arrows). (A) Negative expression of fascin-1 in well-differentiated LSCC. High expression of fascin-1 in (B) well-differentiated, (C) poorly differentiated, (D) supraglottic, (E) glottic, and (F) subglottic LSCC. (G) Negative control staining (phosphate buffered saline [PBS]) of fascin-1. (Magnification 400×, insets 1000×).
    Figure Legend Snippet: Representative immunohistochemistry staining of fascin-1 expression in LSCC. Positive immunostaining of fascin-1 is in cytoplasm of tumor cells (pink arrows) and in endothelial cells of microvessels (blue arrows). (A) Negative expression of fascin-1 in well-differentiated LSCC. High expression of fascin-1 in (B) well-differentiated, (C) poorly differentiated, (D) supraglottic, (E) glottic, and (F) subglottic LSCC. (G) Negative control staining (phosphate buffered saline [PBS]) of fascin-1. (Magnification 400×, insets 1000×).

    Techniques Used: Immunohistochemistry, Staining, Expressing, Immunostaining, Negative Control

    Western blot analysis and quantification of protein levels of fascin-1 (A), ezrin (C) and paxillin (E) in LSCC (L) and ANM (A) tissues (P1-P10). Data are normalized by the loading control GAPDH. Bars are mean±SD fold increase or decrease from experiments performed in triplicate. Arrowheads represent data for each subject.
    Figure Legend Snippet: Western blot analysis and quantification of protein levels of fascin-1 (A), ezrin (C) and paxillin (E) in LSCC (L) and ANM (A) tissues (P1-P10). Data are normalized by the loading control GAPDH. Bars are mean±SD fold increase or decrease from experiments performed in triplicate. Arrowheads represent data for each subject.

    Techniques Used: Western Blot

    2) Product Images from "Fluorescent Arc/Arg3.1 indicator mice: a versatile tool to study brain activity changes in vitro and in vivo"

    Article Title: Fluorescent Arc/Arg3.1 indicator mice: a versatile tool to study brain activity changes in vitro and in vivo

    Journal: Journal of neuroscience methods

    doi: 10.1016/j.jneumeth.2009.07.015

    Increased d4EGFP expression in a mouse model of Alzheimer’s disease Representative images of DG hippocampus (coronal sections) from one month-old Tg Arc/Arg3.1-d4EGFP and Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP littermate mice are shown after visualization of d4EGFP by DAB (A, B) or immunofluorescence (C,D). Note the increased number of and labeling intensity of d4EGFP-positive neurons in CA1 (B) and DG granule cells (B, D) in the Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP mice. Brain sections stained with anti-GFP antibody and visualized by biotinylated (DAB method) or FITC-conjugated secondary antibodies. The fluorescent images are single confocal scans. Abbreviations: db – dorsal blade of DG, vb – ventral blade of DG; scale bars = 250 μm (A,B) and 100 μm (C,D). (E) Count of d4EGFP-positive neurons in Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP mice, as percent change from control Tg Arc/Arg3.1-d4EGFP littermates, in the DG: (% mean ± SD): 216.3 ± 7.7 vs. 100 ± 6.4 (**p
    Figure Legend Snippet: Increased d4EGFP expression in a mouse model of Alzheimer’s disease Representative images of DG hippocampus (coronal sections) from one month-old Tg Arc/Arg3.1-d4EGFP and Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP littermate mice are shown after visualization of d4EGFP by DAB (A, B) or immunofluorescence (C,D). Note the increased number of and labeling intensity of d4EGFP-positive neurons in CA1 (B) and DG granule cells (B, D) in the Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP mice. Brain sections stained with anti-GFP antibody and visualized by biotinylated (DAB method) or FITC-conjugated secondary antibodies. The fluorescent images are single confocal scans. Abbreviations: db – dorsal blade of DG, vb – ventral blade of DG; scale bars = 250 μm (A,B) and 100 μm (C,D). (E) Count of d4EGFP-positive neurons in Tg 5XFAD /Tg Arc/Arg3.1-d4EGFP mice, as percent change from control Tg Arc/Arg3.1-d4EGFP littermates, in the DG: (% mean ± SD): 216.3 ± 7.7 vs. 100 ± 6.4 (**p

    Techniques Used: Expressing, Mouse Assay, Immunofluorescence, Labeling, Staining

    3) Product Images from "Anti-α4β1 Integrin Antibodies Attenuated Brain Inflammatory Changes in a Mouse Model of Alzheimer’s Disease"

    Article Title: Anti-α4β1 Integrin Antibodies Attenuated Brain Inflammatory Changes in a Mouse Model of Alzheimer’s Disease

    Journal: Current Alzheimer Research

    doi: 10.2174/1567205015666180801111033

    Both the isotype control and anti-CD49d antibody injections affected brain CD4 immunoreactivity. C57BL/6 and APP/PS1 mice were injected intravenously (tail-vein) with Saline, IgG isotype control (purified NA/LE Rat IgG2b) (I.C.) or 2 mg/kg purified NA/LE rat anti-mouse CD49d (anti-CD49d) once a week for 4 weeks. Right brain hemispheres from all the mice were fixed and serially sectioned. The presence of IgG2b antibody in the brain was assessed by immunostaining using biotinylated anti-mouse IgG2b antibody (8A) . The presence of T cells in the brain was visualized using anti-CD4 antibody and immunoreactivity observed in the striatum (8B) and parietal cortex (8C) was imaged. Representative images from 3-6 animals per group are shown at 20X magnification with 63X magnification insets.
    Figure Legend Snippet: Both the isotype control and anti-CD49d antibody injections affected brain CD4 immunoreactivity. C57BL/6 and APP/PS1 mice were injected intravenously (tail-vein) with Saline, IgG isotype control (purified NA/LE Rat IgG2b) (I.C.) or 2 mg/kg purified NA/LE rat anti-mouse CD49d (anti-CD49d) once a week for 4 weeks. Right brain hemispheres from all the mice were fixed and serially sectioned. The presence of IgG2b antibody in the brain was assessed by immunostaining using biotinylated anti-mouse IgG2b antibody (8A) . The presence of T cells in the brain was visualized using anti-CD4 antibody and immunoreactivity observed in the striatum (8B) and parietal cortex (8C) was imaged. Representative images from 3-6 animals per group are shown at 20X magnification with 63X magnification insets.

    Techniques Used: Mouse Assay, Injection, Purification, Immunostaining

    4) Product Images from "Immunohistochemical analysis of macroautophagy"

    Article Title: Immunohistochemical analysis of macroautophagy

    Journal: Autophagy

    doi: 10.4161/auto.22968

    Figure 5. The highest expression of LC3A and LC3B in nonstarved control mice was found in brain tissue. ( A ) Western blot analysis of LC3A and LC3B in different tissue lysates. GAPDH served as a loading control. ( B ) Immunohistochemical detection of LC3A and LC3B in mouse brain using the Vectastain ABC system. Tissue samples were isolated from fed control mice. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:3000) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100). Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm.
    Figure Legend Snippet: Figure 5. The highest expression of LC3A and LC3B in nonstarved control mice was found in brain tissue. ( A ) Western blot analysis of LC3A and LC3B in different tissue lysates. GAPDH served as a loading control. ( B ) Immunohistochemical detection of LC3A and LC3B in mouse brain using the Vectastain ABC system. Tissue samples were isolated from fed control mice. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:3000) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100). Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm.

    Techniques Used: Expressing, Mouse Assay, Western Blot, Immunohistochemistry, Isolation, Staining

    Figure 7. Optimal immunohistochemical detection of LC3B requires processing of tissue samples in a suitable fixative. Liver samples were isolated from Gfp-Lc3 tg/tg mice that underwent starvation for 48 h. After fixation in different fixatives for 24 h, tissues were paraffin-embedded and stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) and Vectastain ABC. For formalin-fixed samples, heat-mediated antigen retrieval was performed either in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0). Scale bar, 20 μm.
    Figure Legend Snippet: Figure 7. Optimal immunohistochemical detection of LC3B requires processing of tissue samples in a suitable fixative. Liver samples were isolated from Gfp-Lc3 tg/tg mice that underwent starvation for 48 h. After fixation in different fixatives for 24 h, tissues were paraffin-embedded and stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) and Vectastain ABC. For formalin-fixed samples, heat-mediated antigen retrieval was performed either in citrate buffer (pH 6.0) or in EDTA buffer (pH 8.0). Scale bar, 20 μm.

    Techniques Used: Immunohistochemistry, Isolation, Mouse Assay, Staining

    Figure 4. Liver from autophagy-deficient Atg7 F/F Alb-Cre + mice but not from autophagy-competent Atg7 +/+ Alb-Cre + shows immunohistochemical staining for LC3A and LC3B. Liver samples were isolated from fed mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:100) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The LC3A and LC3B positive area was quantified. ***p
    Figure Legend Snippet: Figure 4. Liver from autophagy-deficient Atg7 F/F Alb-Cre + mice but not from autophagy-competent Atg7 +/+ Alb-Cre + shows immunohistochemical staining for LC3A and LC3B. Liver samples were isolated from fed mice (control) or from mice that underwent starvation for 48 h. After fixation in neutral buffered formalin for 24 h, tissues were paraffin-embedded and stained for LC3A ( A ) and LC3B ( B ) using rabbit polyclonal anti-LC3A (Abgent, 1:100) and biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:100) in combination with Vectastain ABC. Heat-mediated antigen retrieval was performed in citrate buffer (pH 6.0). Scale bar, 20 μm. The LC3A and LC3B positive area was quantified. ***p

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining, Isolation

    Figure 9. LC3B dots are detectable in frozen liver sections using a staining procedure with alkaline phosphatase and are localized on the surface of lipid droplets. Liver samples were isolated from Atg7 +/+ Alb-Cre + ( A ) or Atg7 F/F Alb-Cre + mice ( B ) that were fed normal chow (control) or underwent starvation for 48 h. After fixation in 4% paraformaldehyde, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000) and Vectastain ABC-AP kit, containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase. LC3B-positive dots (arrows) were detected on the surface of lipid droplets. These structures could be stained using oil red O. Scale bar, 20 μm.
    Figure Legend Snippet: Figure 9. LC3B dots are detectable in frozen liver sections using a staining procedure with alkaline phosphatase and are localized on the surface of lipid droplets. Liver samples were isolated from Atg7 +/+ Alb-Cre + ( A ) or Atg7 F/F Alb-Cre + mice ( B ) that were fed normal chow (control) or underwent starvation for 48 h. After fixation in 4% paraformaldehyde, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000) and Vectastain ABC-AP kit, containing biotinylated alkaline phosphatase instead of biotinylated horseradish peroxidase. LC3B-positive dots (arrows) were detected on the surface of lipid droplets. These structures could be stained using oil red O. Scale bar, 20 μm.

    Techniques Used: Staining, Isolation, Mouse Assay

    Figure 8. Immunohistochemical staining of LC3B is enhanced in frozen liver sections. Liver samples were isolated from Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice ( A ) or from transgenic Gfp-Lc3 tg/+ or Gfp-Lc3 tg/tg mice ( B ). Some animals were fed normal chow (control), others underwent starvation for 48 h. After fixation in acetone, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000 [ Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + samples] or 1:30,000 [ Gfp-Lc3 tg/+ and Gfp-Lc3 tg/tg samples]) and Vectastain ABC. Scale bar, 40 μm.
    Figure Legend Snippet: Figure 8. Immunohistochemical staining of LC3B is enhanced in frozen liver sections. Liver samples were isolated from Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + mice ( A ) or from transgenic Gfp-Lc3 tg/+ or Gfp-Lc3 tg/tg mice ( B ). Some animals were fed normal chow (control), others underwent starvation for 48 h. After fixation in acetone, frozen sections were stained for LC3B using biotinylated mouse monoclonal anti-LC3B (clone 5F10, Nanotools, 1:1,000 [ Atg7 +/+ Alb-Cre + and Atg7 F/F Alb-Cre + samples] or 1:30,000 [ Gfp-Lc3 tg/+ and Gfp-Lc3 tg/tg samples]) and Vectastain ABC. Scale bar, 40 μm.

    Techniques Used: Immunohistochemistry, Staining, Isolation, Mouse Assay, Transgenic Assay

    5) Product Images from "Knockdown of L Calcium Channel Subtypes: Differential Effects in Neuropathic Pain"

    Article Title: Knockdown of L Calcium Channel Subtypes: Differential Effects in Neuropathic Pain

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3145-09.2010

    Penetration of TP10-PNAs restricted to the lumbar spinal dorsal horn. A , Naive animals received an intrathecal injection of either biotin-coupled transportan 10-anti-Ca V 1.2 PNA construct ( Aa , Ab , Ad ), or vehicle ( Ac ). Then, peroxidase activity was revealed in the dorsal horn of the L5 lumbar ( Aa , Ac ) and Th8-10 thoracic ( Ab ) spinal cord, and in the L5 DRG ( Ad ). A positive signal was seen only in L5 dorsal horn after injection of biotinylated TP10-PNA constructs (black star in Aa ). No signal was observed in thoracic segments (open star in Ab ), or after vehicle injection (open star in Ac ). Sensory neurons in the L5 DRG also remained unlabeled (arrows in Ad ). Sections were observed from three animals in each condition (biotinylated PNA- or vehicle-injected animals). B , Confocal views of ipsilateral DRG sections from SNL rats, before ( Ba ) or after ( Bb ) intrathecal application of TP10-anti-Ca V 1.2 PNA construct. Immunofluorescence of individual neurons is similar in both conditions (arrows, see quantification in supplemental Fig. S1 B as supplemental material). Scale bars: 50 μm in A , 20 μm in B .
    Figure Legend Snippet: Penetration of TP10-PNAs restricted to the lumbar spinal dorsal horn. A , Naive animals received an intrathecal injection of either biotin-coupled transportan 10-anti-Ca V 1.2 PNA construct ( Aa , Ab , Ad ), or vehicle ( Ac ). Then, peroxidase activity was revealed in the dorsal horn of the L5 lumbar ( Aa , Ac ) and Th8-10 thoracic ( Ab ) spinal cord, and in the L5 DRG ( Ad ). A positive signal was seen only in L5 dorsal horn after injection of biotinylated TP10-PNA constructs (black star in Aa ). No signal was observed in thoracic segments (open star in Ab ), or after vehicle injection (open star in Ac ). Sensory neurons in the L5 DRG also remained unlabeled (arrows in Ad ). Sections were observed from three animals in each condition (biotinylated PNA- or vehicle-injected animals). B , Confocal views of ipsilateral DRG sections from SNL rats, before ( Ba ) or after ( Bb ) intrathecal application of TP10-anti-Ca V 1.2 PNA construct. Immunofluorescence of individual neurons is similar in both conditions (arrows, see quantification in supplemental Fig. S1 B as supplemental material). Scale bars: 50 μm in A , 20 μm in B .

    Techniques Used: Injection, Construct, Activity Assay, Immunofluorescence

    6) Product Images from "Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice"

    Article Title: Oxidized Phospholipids are Proinflammatory and Proatherogenic in Hypercholesterolemic Mice

    Journal: Nature

    doi: 10.1038/s41586-018-0198-8

    Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.
    Figure Legend Snippet: Plasma E06-scFv binds to atherosclerotic lesions and apoptotic thymocytes and is present in aorta of Ldlr −/− /E06-scFv mice a , Staining of atherosclerotic lesions of WHHL rabbit aorta with E06-scFv plasma (left panel), and Ldlr −/− mice (right panel) (both at dilution of 1:20), visualized using biotinylated anti-Myc mAb and ABC-AP VectaStain kit. b , Deconvolution microscopy of E06-scFv plasma (1:20 dilution) binding to apoptotic but not normal cells. Blue, nuclei stained with Hoechst dye; Green, FITC-labeled anti-His tag mAb; Red, Annexin V-PE. c , Binding of E06-scFv plasma (1:20 dilution) to apoptotic thymocytes (7AAD+/Annexin V+) by FACS analysis. d , Expression of E06-scFv in aortic lesion of Ldlr −/− / E06-scFv but not Ldlr −/− mouse. Cross-sections at the AV were stained with biotinylated anti-Myc mAb to identify presence of E06-scFv in atherosclerotic lesion. Nuclei counterstained using Hematoxylin QS (Original ×200). Panels a-c are representative of similar studies with 5 other plasma samples. Panel d is representative of studies in 3 other aortic sections.

    Techniques Used: Mouse Assay, Staining, Microscopy, Binding Assay, Labeling, FACS, Expressing

    7) Product Images from "Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury"

    Article Title: Aging impairs induction of redox factor-1 after heat stress: a potential mechanism for heat-induced liver injury

    Journal: International Journal of Physiology, Pathophysiology and Pharmacology

    doi:

    Colocalization of Ref-1 and Trx-1 within ductular reactions of aged animals. A: Ductular reaction (DR) in an old rat showing positive staining for Ref-1. The arrow indicates a cell positive for Ref-1. Multiple nuclei within bile ducts cells are positive for Ref-1. B: Ductular reaction in an old rat showing positive staining for Trx-1. The arrow indicates a cell positive for Trx-1. Frequent positive staining for Trx-1 was observed in cells within DR. Sections in Panels A and B were counterstained with hematoxylin. C: Colocalization of Ref-1 (black) and Trx-1 (brown) in cells of a DR. Arrowheads indicate cells positive for both Ref-1 and Trx-1. D: Colocalization of Ref-1 (black) and β-catenin (brown) in biliary epithelial cells of a DR. Arrowheads indicate cells positive for both Ref-1 and β-catenin. Sections in Panels C and D were not counterstained. Scale bar = 100 μm.
    Figure Legend Snippet: Colocalization of Ref-1 and Trx-1 within ductular reactions of aged animals. A: Ductular reaction (DR) in an old rat showing positive staining for Ref-1. The arrow indicates a cell positive for Ref-1. Multiple nuclei within bile ducts cells are positive for Ref-1. B: Ductular reaction in an old rat showing positive staining for Trx-1. The arrow indicates a cell positive for Trx-1. Frequent positive staining for Trx-1 was observed in cells within DR. Sections in Panels A and B were counterstained with hematoxylin. C: Colocalization of Ref-1 (black) and Trx-1 (brown) in cells of a DR. Arrowheads indicate cells positive for both Ref-1 and Trx-1. D: Colocalization of Ref-1 (black) and β-catenin (brown) in biliary epithelial cells of a DR. Arrowheads indicate cells positive for both Ref-1 and β-catenin. Sections in Panels C and D were not counterstained. Scale bar = 100 μm.

    Techniques Used: Staining

    Increase in Trx-1 abundance in old animals after hyperthermia. Left panel: representative immunoblot of Trx-1 in young and old animals under control conditions, and at the indicated times after heat stress (in hours). Ponceau stained membrane (Pon) demonstrates equal loading and transfer. Right panel: quantitation of Trx-1 expression, normalized to the Ponceau stain and further normalized to the young, nonheated group. †Significant effect of heat stress (n = 7-9 young and old animals in the nonheated condition, and at each time point after heat stress).
    Figure Legend Snippet: Increase in Trx-1 abundance in old animals after hyperthermia. Left panel: representative immunoblot of Trx-1 in young and old animals under control conditions, and at the indicated times after heat stress (in hours). Ponceau stained membrane (Pon) demonstrates equal loading and transfer. Right panel: quantitation of Trx-1 expression, normalized to the Ponceau stain and further normalized to the young, nonheated group. †Significant effect of heat stress (n = 7-9 young and old animals in the nonheated condition, and at each time point after heat stress).

    Techniques Used: Staining, Quantitation Assay, Expressing

    Localization of hepatic Trx-1. A: Trx-1 staining in a young, control liver. An identical pattern of cellular localization was observed in the old control rats (not shown). The arrowhead indicates positive staining in a bile duct, arrows indicate positive staining in nonparenchymal cells. B: Colocalization of Trx-1 (black) and HO-1 (brown) in a young control rat; double-positive cells are indicated by arrowheads (results are representative of 3 separate young and old control animals, with the experiment run in triplicate). C: Colocalization of Trx-1 (brown) and iron deposits (cyan) in an old, control rat, double positive cells are indicated by arrowheads (results are representative of 3 separate old animals). Scale bar = 100 μm.
    Figure Legend Snippet: Localization of hepatic Trx-1. A: Trx-1 staining in a young, control liver. An identical pattern of cellular localization was observed in the old control rats (not shown). The arrowhead indicates positive staining in a bile duct, arrows indicate positive staining in nonparenchymal cells. B: Colocalization of Trx-1 (black) and HO-1 (brown) in a young control rat; double-positive cells are indicated by arrowheads (results are representative of 3 separate young and old control animals, with the experiment run in triplicate). C: Colocalization of Trx-1 (brown) and iron deposits (cyan) in an old, control rat, double positive cells are indicated by arrowheads (results are representative of 3 separate old animals). Scale bar = 100 μm.

    Techniques Used: Staining

    Immunolocalization of Ref-1 in the liver. A: Immunostaining for Ref-1 alone (brown), with hematoxylin counterstain. The arrow indicates a hepatocyte nucleus positive for Ref-1; the arrowhead indicates a sinusoidal lining cell positive for Ref-1. Positive staining was also apparent in cells lining the bile duct (BD). B: Double staining for Ref-1 (black) and glutamine synthetase (GS; brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in hepatocytes; the arrow indicates a hepatocyte nucleus positive for Ref-1. HV: terminal hepatic venule C: Double staining for Ref-1 (black) and β-catenin (brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in biliary epithelial cells; the arrow indicates a hepatocyte nucleus positive for Ref-1. D: Double staining for Ref-1 (black) and Trx-1 (brown). Arrowheads indicate colocalization in bile ducts (BD); double arrows indicate colocalization in sinusoidal lining cells (SEC; Kupffer cells). The arrow indicates a hepatocyte positive for Ref-1. Sections in panels B-D were not counterstained. All panels are images from young, control rats and are representative of 5 individual animals. The cellular localization of Ref-1 was identical in old, nonheated rats (not shown). Scale bar = 100 μm.
    Figure Legend Snippet: Immunolocalization of Ref-1 in the liver. A: Immunostaining for Ref-1 alone (brown), with hematoxylin counterstain. The arrow indicates a hepatocyte nucleus positive for Ref-1; the arrowhead indicates a sinusoidal lining cell positive for Ref-1. Positive staining was also apparent in cells lining the bile duct (BD). B: Double staining for Ref-1 (black) and glutamine synthetase (GS; brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in hepatocytes; the arrow indicates a hepatocyte nucleus positive for Ref-1. HV: terminal hepatic venule C: Double staining for Ref-1 (black) and β-catenin (brown). Colocalization (indicated by arrowhead) demonstrates Ref-1 expression in biliary epithelial cells; the arrow indicates a hepatocyte nucleus positive for Ref-1. D: Double staining for Ref-1 (black) and Trx-1 (brown). Arrowheads indicate colocalization in bile ducts (BD); double arrows indicate colocalization in sinusoidal lining cells (SEC; Kupffer cells). The arrow indicates a hepatocyte positive for Ref-1. Sections in panels B-D were not counterstained. All panels are images from young, control rats and are representative of 5 individual animals. The cellular localization of Ref-1 was identical in old, nonheated rats (not shown). Scale bar = 100 μm.

    Techniques Used: Immunostaining, Staining, Double Staining, Expressing, Size-exclusion Chromatography

    8) Product Images from "GM1 ganglioside-independent intoxication by Cholera toxin"

    Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006862

    CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p
    Figure Legend Snippet: CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p

    Techniques Used: Incubation

    GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.
    Figure Legend Snippet: GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.

    Techniques Used: Inhibition, Cell Culture, Staining, CtB Assay, Fluorescence, Flow Cytometry, Cytometry, Luminescence Assay, Polyacrylamide Gel Electrophoresis

    9) Product Images from "Evaluation of the soft tissue biocompatibility of MgCa0.8 and surgical steel 316L in vivo: a comparative study in rabbits"

    Article Title: Evaluation of the soft tissue biocompatibility of MgCa0.8 and surgical steel 316L in vivo: a comparative study in rabbits

    Journal: BioMedical Engineering OnLine

    doi: 10.1186/1475-925X-9-63

    Image sections of immunohistochemically stained cranial tibial muscle cross sections . (A) Periimplant fibrous tissue of an MgCa0.8 screw six weeks post operatively (CD79α, 400×). Arrows represent CD79α positive B-lymphocytes. (B) Periimplant fibrous tissue of an S316L screw two weeks postoperatively (CD3, 400×). Triangles indicate CD3 positive T-lymphocytes.
    Figure Legend Snippet: Image sections of immunohistochemically stained cranial tibial muscle cross sections . (A) Periimplant fibrous tissue of an MgCa0.8 screw six weeks post operatively (CD79α, 400×). Arrows represent CD79α positive B-lymphocytes. (B) Periimplant fibrous tissue of an S316L screw two weeks postoperatively (CD3, 400×). Triangles indicate CD3 positive T-lymphocytes.

    Techniques Used: Staining

    10) Product Images from "Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey"

    Article Title: Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhn078

    Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).
    Figure Legend Snippet: Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).

    Techniques Used: Labeling, Immunohistochemistry

    11) Product Images from "Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey"

    Article Title: Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhn078

    GAD67 labeling patterns in coronal sections of monkey fetal telencephalon at E55 ( A , a , a ′), E68 ( B , b ) and E75 ( C , c , c ′). ( A ) At E55 as at earlier stage of cortical development, GAD67-containing neurons were mainly present in the MZ and below the CP in the upper part of the subplate (SPu). In the MZ these neurons were surrounded by dense plexus of stained fibers as illustrated at higher magnification (inset a ′). Some neurons were also observed in the IZ (arrows). Many of them in this layer displayed a unipolar migratory-like morphology (inset a). ( B ) At E68 a general increased density of GAD67-labeled neurons was observed but this increase was particularly striking in the lower part of the subplate (SPL) (arrows). Inset b: In this later layer, many GAD67-labeled cells corresponded to morphologically differentiated neurons, that is, neurons with strong labeling of the cell body and proximal dendrites (arrow). These neurons contrast with migratory-like postmitotic neurons lightly labeled for GAD67 (arrowhead). ( C ) At E75 the thickness of the SP and the density of GAD67-containing neurons further increased. More GAD67-labeled neurons were also observed in the MZ and the IZ. Some GAD67-containing neurons were now present in the CP (arrows). Higher magnification illustrating morphologically differentiated neurons and axon terminals labeled for GAD67 in the SP (inset c) and in the CP (inset c ′). ( D ) At E75 Adjacent section stained for cresyl violet showed that the SP is at this age up to 3 times thicker than the CP. Scale bars = 200 μm ( A – D ), 50 μm (a′, b, c, c′), 25 μm ( a ).
    Figure Legend Snippet: GAD67 labeling patterns in coronal sections of monkey fetal telencephalon at E55 ( A , a , a ′), E68 ( B , b ) and E75 ( C , c , c ′). ( A ) At E55 as at earlier stage of cortical development, GAD67-containing neurons were mainly present in the MZ and below the CP in the upper part of the subplate (SPu). In the MZ these neurons were surrounded by dense plexus of stained fibers as illustrated at higher magnification (inset a ′). Some neurons were also observed in the IZ (arrows). Many of them in this layer displayed a unipolar migratory-like morphology (inset a). ( B ) At E68 a general increased density of GAD67-labeled neurons was observed but this increase was particularly striking in the lower part of the subplate (SPL) (arrows). Inset b: In this later layer, many GAD67-labeled cells corresponded to morphologically differentiated neurons, that is, neurons with strong labeling of the cell body and proximal dendrites (arrow). These neurons contrast with migratory-like postmitotic neurons lightly labeled for GAD67 (arrowhead). ( C ) At E75 the thickness of the SP and the density of GAD67-containing neurons further increased. More GAD67-labeled neurons were also observed in the MZ and the IZ. Some GAD67-containing neurons were now present in the CP (arrows). Higher magnification illustrating morphologically differentiated neurons and axon terminals labeled for GAD67 in the SP (inset c) and in the CP (inset c ′). ( D ) At E75 Adjacent section stained for cresyl violet showed that the SP is at this age up to 3 times thicker than the CP. Scale bars = 200 μm ( A – D ), 50 μm (a′, b, c, c′), 25 μm ( a ).

    Techniques Used: Labeling, Staining

    12) Product Images from "Selective Restoration of Pomc Expression in Glutamatergic POMC Neurons: Evidence for a Dynamic Hypothalamic Neurotransmitter Network"

    Article Title: Selective Restoration of Pomc Expression in Glutamatergic POMC Neurons: Evidence for a Dynamic Hypothalamic Neurotransmitter Network

    Journal: eNeuro

    doi: 10.1523/ENEURO.0400-18.2019

    IHC for POMC cell counts in control and restored mice, and from VGlut2-Cre; tdTomato animals. A , POMC-IR in a male control mouse detected with an Alexa Fluor 568 (red) secondary antibody (1:500). B , POMC-IR in a male restored mouse detected with an Alexa Fluor 568 (red) secondary antibody (1:500). C , POMC-IR in a female control mouse detected with biotinylated secondary antibody (1:500) and visualized with a diaminobenzidine (DAB) reaction (brown). D , POMC-IR in a female restored mouse detected with biotinylated secondary antibody (1:500) and visualized with a DAB reaction (brown). E , POMC-IR in a female Vglut2-Cre; tdTomato mouse detected with an Alexa Fluor 488 (green) secondary antibody (1:500; mirrored section from Fig. 1 G , H ). F , POMC neuron cell counts from sections (three per mouse). There was no difference between control (blue bars) or restored (green bars) mice, but only in the method of secondary labeling used. Male data for each group represented by filled blue circles and female data shown by filled pink circles; **** p
    Figure Legend Snippet: IHC for POMC cell counts in control and restored mice, and from VGlut2-Cre; tdTomato animals. A , POMC-IR in a male control mouse detected with an Alexa Fluor 568 (red) secondary antibody (1:500). B , POMC-IR in a male restored mouse detected with an Alexa Fluor 568 (red) secondary antibody (1:500). C , POMC-IR in a female control mouse detected with biotinylated secondary antibody (1:500) and visualized with a diaminobenzidine (DAB) reaction (brown). D , POMC-IR in a female restored mouse detected with biotinylated secondary antibody (1:500) and visualized with a DAB reaction (brown). E , POMC-IR in a female Vglut2-Cre; tdTomato mouse detected with an Alexa Fluor 488 (green) secondary antibody (1:500; mirrored section from Fig. 1 G , H ). F , POMC neuron cell counts from sections (three per mouse). There was no difference between control (blue bars) or restored (green bars) mice, but only in the method of secondary labeling used. Male data for each group represented by filled blue circles and female data shown by filled pink circles; **** p

    Techniques Used: Immunohistochemistry, Mouse Assay, Labeling

    13) Product Images from "Induction of intestinalization in human esophageal keratinocytes is a multistep process"

    Article Title: Induction of intestinalization in human esophageal keratinocytes is a multistep process

    Journal: Carcinogenesis

    doi: 10.1093/carcin/bgn227

    Senescence checkpoint genes are activated by chromatin remodeling agents 5-AzaC in esophageal keratinocytes. After 5-AzaC treatment, EPC-hTERT.D1-Cdx2 and MIGR1 (EPC.D1.X2 and EPC.D1.M, respectively) control cells exhibit characteristics of senescence including altered morphology, increased cell volume and subconfluent growth arrest in the normal medium. ( A ) Morphology of EPC-hTERT.D1.Cdx2 and control EPC-hTERT.D1.MIGR1 cells after 5AzaC at 1 and 5 μM for 5 days. Induction of senescence-associated β-galactosidase activity was identified by staining (blue). ( B ) 5-AzaC treatment induces the expression of senescence-associated p16(INK4a) and p21(Waf1/Cip1). Western Blot for p16 and p21 after cells were treated with 1 μM 5-AzaC or control diluent for 5 days. β-Actin served as an internal loading control. ( C ) Proposed model for the early molecular events preceding the onset of intestinalization in esophageal keratinocytes.
    Figure Legend Snippet: Senescence checkpoint genes are activated by chromatin remodeling agents 5-AzaC in esophageal keratinocytes. After 5-AzaC treatment, EPC-hTERT.D1-Cdx2 and MIGR1 (EPC.D1.X2 and EPC.D1.M, respectively) control cells exhibit characteristics of senescence including altered morphology, increased cell volume and subconfluent growth arrest in the normal medium. ( A ) Morphology of EPC-hTERT.D1.Cdx2 and control EPC-hTERT.D1.MIGR1 cells after 5AzaC at 1 and 5 μM for 5 days. Induction of senescence-associated β-galactosidase activity was identified by staining (blue). ( B ) 5-AzaC treatment induces the expression of senescence-associated p16(INK4a) and p21(Waf1/Cip1). Western Blot for p16 and p21 after cells were treated with 1 μM 5-AzaC or control diluent for 5 days. β-Actin served as an internal loading control. ( C ) Proposed model for the early molecular events preceding the onset of intestinalization in esophageal keratinocytes.

    Techniques Used: Activity Assay, Staining, Expressing, Western Blot

    14) Product Images from "Trafficking to the Plasma Membrane of the Seven-Transmembrane Protein Encoded by Human Herpesvirus 6 U51 Gene Involves a Cell-Specific Function Present in T Lymphocytes"

    Article Title: Trafficking to the Plasma Membrane of the Seven-Transmembrane Protein Encoded by Human Herpesvirus 6 U51 Gene Involves a Cell-Specific Function Present in T Lymphocytes

    Journal: Journal of Virology

    doi:

    Synthesis of U51 in CBMCs infected with HHV-6(A)U1102 (A) or HHV-6(B)Z29 (B and C). (A and B) Detection of U51 by metabolic labeling with [ 35 S]methionine and [ 35 S]cysteine ( 35 S-met). (C) Detection of cell surface expression by labeling with biotin prior to harvesting (biotin). Immunoprecipitations were performed on lysates of infected (+) or uninfected (−) CBMCs with immune serum 6 (Immune) or with preimmune serum (Pre). Note that U51 (black arrowheads) was detectable as radiolabeled protein in lanes d and h and as biotinylated species in lane l. Numbers at left show molecular mass in kilodaltons.
    Figure Legend Snippet: Synthesis of U51 in CBMCs infected with HHV-6(A)U1102 (A) or HHV-6(B)Z29 (B and C). (A and B) Detection of U51 by metabolic labeling with [ 35 S]methionine and [ 35 S]cysteine ( 35 S-met). (C) Detection of cell surface expression by labeling with biotin prior to harvesting (biotin). Immunoprecipitations were performed on lysates of infected (+) or uninfected (−) CBMCs with immune serum 6 (Immune) or with preimmune serum (Pre). Note that U51 (black arrowheads) was detectable as radiolabeled protein in lanes d and h and as biotinylated species in lane l. Numbers at left show molecular mass in kilodaltons.

    Techniques Used: Infection, Labeling, Expressing

    Immunofluorescence staining of U51 in HHV-6(B)Z29-infected CBMCs. Cells fixed with acetone were reacted with immune serum 6, followed by biotinylated anti-mouse antibodies and Extravidin coupled to TRITC.
    Figure Legend Snippet: Immunofluorescence staining of U51 in HHV-6(B)Z29-infected CBMCs. Cells fixed with acetone were reacted with immune serum 6, followed by biotinylated anti-mouse antibodies and Extravidin coupled to TRITC.

    Techniques Used: Immunofluorescence, Staining, Infection

    Lack of cell surface expression of U51 in transfected 143tk− cells detected by biotinylation. VacT7-preinfected 143tk− cells were transfected with p51-HA (lanes b and e), cotransfected with p51-HA and pgD (lanes c and f), or mock transfected (lanes a and d). Cells were metabolically labeled with [ 35 S]methionine and [ 35 S]cysteine and surface labeled with biotin immediately prior to harvesting. U51 and gD were immunoprecipitated, separated by electrophoresis, and transferred to a nitrocellulose sheet. (A) Autoradiographic image. (B) Avidin-peroxidase staining of biotinylated proteins. Note that U51 is detectable only as radiolabeled species (black arrowhead). gD is detectable both as radiolabeled and as biotinylated species (white arrowheads). Numbers at left show molecular mass in kilodaltons.
    Figure Legend Snippet: Lack of cell surface expression of U51 in transfected 143tk− cells detected by biotinylation. VacT7-preinfected 143tk− cells were transfected with p51-HA (lanes b and e), cotransfected with p51-HA and pgD (lanes c and f), or mock transfected (lanes a and d). Cells were metabolically labeled with [ 35 S]methionine and [ 35 S]cysteine and surface labeled with biotin immediately prior to harvesting. U51 and gD were immunoprecipitated, separated by electrophoresis, and transferred to a nitrocellulose sheet. (A) Autoradiographic image. (B) Avidin-peroxidase staining of biotinylated proteins. Note that U51 is detectable only as radiolabeled species (black arrowhead). gD is detectable both as radiolabeled and as biotinylated species (white arrowheads). Numbers at left show molecular mass in kilodaltons.

    Techniques Used: Expressing, Transfection, Metabolic Labelling, Labeling, Immunoprecipitation, Electrophoresis, Avidin-Biotin Assay, Staining

    15) Product Images from "Abnormalities of Thymic Stroma may Contribute to Immune Dysregulation in Murine Models of Leaky Severe Combined Immunodeficiency"

    Article Title: Abnormalities of Thymic Stroma may Contribute to Immune Dysregulation in Murine Models of Leaky Severe Combined Immunodeficiency

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2011.00015

    Altered distribution of thymic DCs populations in lig4 R/R and rag1 S/S thymuses . Top panels: FACS dot plot analysis of the distribution of CD11c + CD45RA − cDCs and CD11c int CD45RA hi pDCs in the thymus of WT, lig4 R/R , and rag1 S/S mice after gating on a population of stromal cells positive for CD11c expression but negative for a cocktail of biotinylated markers specific for markers of T and B lymphocytes, erythroid, granulocyte, and macrophage lineages. Lower panels: Proportion of thymic cDCs and pDCs in WT, lig4 R/R , and rag1 S/S mice. Mean values ± SE are shown. At least six mice per group were analyzed.
    Figure Legend Snippet: Altered distribution of thymic DCs populations in lig4 R/R and rag1 S/S thymuses . Top panels: FACS dot plot analysis of the distribution of CD11c + CD45RA − cDCs and CD11c int CD45RA hi pDCs in the thymus of WT, lig4 R/R , and rag1 S/S mice after gating on a population of stromal cells positive for CD11c expression but negative for a cocktail of biotinylated markers specific for markers of T and B lymphocytes, erythroid, granulocyte, and macrophage lineages. Lower panels: Proportion of thymic cDCs and pDCs in WT, lig4 R/R , and rag1 S/S mice. Mean values ± SE are shown. At least six mice per group were analyzed.

    Techniques Used: FACS, Mouse Assay, Expressing

    16) Product Images from "Mitogen- and stress-activated protein kinase 1-induced neuroprotection in Huntington's disease: role on chromatin remodeling at the PGC-1-alpha promoter"

    Article Title: Mitogen- and stress-activated protein kinase 1-induced neuroprotection in Huntington's disease: role on chromatin remodeling at the PGC-1-alpha promoter

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddr148

    MSK-1-expressing cells exhibit DARPP-32 immunoreactivity in the area surrounding the injection point 10 weeks post-infection. ( A ) Rats were infected as indicated in Fig. 1 . ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels), 10 weeks later. Note the down-regulation of both MSK-1 and DARPP-32 in the right striatum infected with LV-Exp-Ht alone. Note also the preserved expression of both MSK-1 and DARPP-32 around the injection point on the left side infected with both LV-Exp-Htt and LV-MSK-1. The right panels show a higher magnification of the area surrounding the LV-Exp + MSK-1 injection point (see square box in the middle panels). Note that all three markers, Exp-Ht, MSK-1 and DARPP-32, are expressed in this area (white arrow). ( C ) The Exp-Htt volume expression in the rostro-caudal extension of the striatum and ( D ) the DARPP-32 loss/Exp-Htt infected area were measured as described in Fig. 1 . Statistics: means ± SEM; eight rats per group. ns, not significant; * P
    Figure Legend Snippet: MSK-1-expressing cells exhibit DARPP-32 immunoreactivity in the area surrounding the injection point 10 weeks post-infection. ( A ) Rats were infected as indicated in Fig. 1 . ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels), 10 weeks later. Note the down-regulation of both MSK-1 and DARPP-32 in the right striatum infected with LV-Exp-Ht alone. Note also the preserved expression of both MSK-1 and DARPP-32 around the injection point on the left side infected with both LV-Exp-Htt and LV-MSK-1. The right panels show a higher magnification of the area surrounding the LV-Exp + MSK-1 injection point (see square box in the middle panels). Note that all three markers, Exp-Ht, MSK-1 and DARPP-32, are expressed in this area (white arrow). ( C ) The Exp-Htt volume expression in the rostro-caudal extension of the striatum and ( D ) the DARPP-32 loss/Exp-Htt infected area were measured as described in Fig. 1 . Statistics: means ± SEM; eight rats per group. ns, not significant; * P

    Techniques Used: Expressing, Injection, Infection

    MSK-1 overexpression in the striatum induces histone H3 and CREB hyperphosphorylation. Rat striata were infected with LV-Htt or LV-Exp-Htt, with or without MSK-1. ( A ) Immunohistochemical detection of phospho-Ser 10 -H3 was performed 4 weeks later with an anti-rabbit Cy3-coupled secondary antibody (upper panels). MSK-1 expression was detected after co-labeling with an anti-mouse anti-FITC secondary antibody, on the same section (lower panels). Note that a majority of cells express P-H3 in the LV-Htt + MSK-1- and LV-Exp-Htt + MSK-1-infected striata but not in the striata infected with LV-Htt or LV-Exp-Htt alone. ( B ) Immunohistochemical detection of phospho-Ser 133 -CREB was performed 4 weeks post-infection. MSK-1 expression was detected as indicated in (A). Note that MSK-1 over-expression enhanced P-CREB immunoreactivity in Htt + MSK-1- and Exp-Htt + MSK-1-infected striata. ( C ) MSK-1, ( D ) P-H3- and ( E ) P-CREB-immunoreactive neurons were counted in the infected areas of the striatum, as indicated in Figure 3 . * P
    Figure Legend Snippet: MSK-1 overexpression in the striatum induces histone H3 and CREB hyperphosphorylation. Rat striata were infected with LV-Htt or LV-Exp-Htt, with or without MSK-1. ( A ) Immunohistochemical detection of phospho-Ser 10 -H3 was performed 4 weeks later with an anti-rabbit Cy3-coupled secondary antibody (upper panels). MSK-1 expression was detected after co-labeling with an anti-mouse anti-FITC secondary antibody, on the same section (lower panels). Note that a majority of cells express P-H3 in the LV-Htt + MSK-1- and LV-Exp-Htt + MSK-1-infected striata but not in the striata infected with LV-Htt or LV-Exp-Htt alone. ( B ) Immunohistochemical detection of phospho-Ser 133 -CREB was performed 4 weeks post-infection. MSK-1 expression was detected as indicated in (A). Note that MSK-1 over-expression enhanced P-CREB immunoreactivity in Htt + MSK-1- and Exp-Htt + MSK-1-infected striata. ( C ) MSK-1, ( D ) P-H3- and ( E ) P-CREB-immunoreactive neurons were counted in the infected areas of the striatum, as indicated in Figure 3 . * P

    Techniques Used: Over Expression, Infection, Immunohistochemistry, Expressing, Labeling

    Exp-Htt-induced down-regulation of DARPP-32 is attenuated by MSK-1 overexpression 4 weeks post-infection. ( A ) The left striatum was infected with a lentiviral construct containing a gene fragment of huntingtin with 82 polyglutamine repeats (Exp-Htt) and the right striatum was infected with LV-Exp-Htt and another lentiviral construct containing the MSK-1 gene (Exp-Htt + MSK-1). ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels) was performed with specific antibodies (see Materials and Methods), 4 weeks after infection. ( C ) The volume of striatal tissue showing Exp-Htt expression or DARPP-32 loss was measured on adjacent coronal sections across the rostro-caudal extension of the striatum. Seven to 10 sections per rat were analyzed (each section was 25 µm thick and was separated by 175 µm). ( D ) Graphical representation of the Exp-Htt volume expression in the rostro-caudal extension of the striatum. ( E ) Graphical representation of DARPP-32 loss/Exp-Ht infected area. Statistics: means ± SEM; eight rats per group). ns, not significant; ** P
    Figure Legend Snippet: Exp-Htt-induced down-regulation of DARPP-32 is attenuated by MSK-1 overexpression 4 weeks post-infection. ( A ) The left striatum was infected with a lentiviral construct containing a gene fragment of huntingtin with 82 polyglutamine repeats (Exp-Htt) and the right striatum was infected with LV-Exp-Htt and another lentiviral construct containing the MSK-1 gene (Exp-Htt + MSK-1). ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels) was performed with specific antibodies (see Materials and Methods), 4 weeks after infection. ( C ) The volume of striatal tissue showing Exp-Htt expression or DARPP-32 loss was measured on adjacent coronal sections across the rostro-caudal extension of the striatum. Seven to 10 sections per rat were analyzed (each section was 25 µm thick and was separated by 175 µm). ( D ) Graphical representation of the Exp-Htt volume expression in the rostro-caudal extension of the striatum. ( E ) Graphical representation of DARPP-32 loss/Exp-Ht infected area. Statistics: means ± SEM; eight rats per group). ns, not significant; ** P

    Techniques Used: Over Expression, Infection, Construct, Expressing

    PGC-1α expression is induced by MSK-1 overexpression in the striatum. LV-Exp-Htt was injected in the left striatum (left panel) and both LV-Exp-Htt and LV-MSK-1 in the right striatum (right panel). Double staining for MSK-1 (red) and PGC-1α (green) was performed. At 4 weeks ( A ) and 10 weeks ( B ) post-infection, Exp-Htt down-regulated the expression of both MSK-1 and PGC-1α (left panels, asterisk). (A and B) In the co-infected sides, MSK-1 overexpression was associated with an increase in PGC-1α immunoreactivity at 4 weeks (A , right panels) and 10 weeks in the area surrounding the injection point ( B , right panels, triangle). ( C and D ) PGC-1α-immunoreactive neurons were counted in the infected area of the striatum at 4 (C) and 10 weeks (D), using the endogenous basal level of PGC-1α expression as threshold. Results are means ± SEM, eights rats per group; *** P
    Figure Legend Snippet: PGC-1α expression is induced by MSK-1 overexpression in the striatum. LV-Exp-Htt was injected in the left striatum (left panel) and both LV-Exp-Htt and LV-MSK-1 in the right striatum (right panel). Double staining for MSK-1 (red) and PGC-1α (green) was performed. At 4 weeks ( A ) and 10 weeks ( B ) post-infection, Exp-Htt down-regulated the expression of both MSK-1 and PGC-1α (left panels, asterisk). (A and B) In the co-infected sides, MSK-1 overexpression was associated with an increase in PGC-1α immunoreactivity at 4 weeks (A , right panels) and 10 weeks in the area surrounding the injection point ( B , right panels, triangle). ( C and D ) PGC-1α-immunoreactive neurons were counted in the infected area of the striatum at 4 (C) and 10 weeks (D), using the endogenous basal level of PGC-1α expression as threshold. Results are means ± SEM, eights rats per group; *** P

    Techniques Used: Pyrolysis Gas Chromatography, Expressing, Over Expression, Injection, Double Staining, Infection

    MSK-1 overexpression prevents Exp-Htt-induced neuronal dysfunction. ( A ) LV-Exp-Htt was injected in the left striatum and both LV-Exp-Htt and LV-MSK-1 were injected in the right striatum. Double staining for MSK-1 (red) and NeuN (green) was performed 10 weeks later. Left panels (Exp-Htt): note the disappearance of both MSK-1 and NeuN labeling within the dotted lines (injection point). Right panels (Exp-Htt + MSK-1): note the down-regulation of both markers within the dotted lines, despite the presence of some immunoreactive cells. Note also the overexpression of MSK-1 and NeuN in the area surrounding the injection point (outside the dotted lines). ( B ) Confocal micrograph showing the co-localization of aggregated Exp-Htt (green) and MSK-1 (red) in the area surrounding the injection point. ( C ) Cells overexpressing NeuN were quantified by image analysis, with a threshold based on mean endogenous NeuN expression. Cells exhibiting a fluorescence level above this threshold were considered positive, and were quantified in eight animals. ( D ) The mean nuclear area of cells in the area surrounding the injection point was analyzed with Hoechst staining and Image-ProPlus software. ** P
    Figure Legend Snippet: MSK-1 overexpression prevents Exp-Htt-induced neuronal dysfunction. ( A ) LV-Exp-Htt was injected in the left striatum and both LV-Exp-Htt and LV-MSK-1 were injected in the right striatum. Double staining for MSK-1 (red) and NeuN (green) was performed 10 weeks later. Left panels (Exp-Htt): note the disappearance of both MSK-1 and NeuN labeling within the dotted lines (injection point). Right panels (Exp-Htt + MSK-1): note the down-regulation of both markers within the dotted lines, despite the presence of some immunoreactive cells. Note also the overexpression of MSK-1 and NeuN in the area surrounding the injection point (outside the dotted lines). ( B ) Confocal micrograph showing the co-localization of aggregated Exp-Htt (green) and MSK-1 (red) in the area surrounding the injection point. ( C ) Cells overexpressing NeuN were quantified by image analysis, with a threshold based on mean endogenous NeuN expression. Cells exhibiting a fluorescence level above this threshold were considered positive, and were quantified in eight animals. ( D ) The mean nuclear area of cells in the area surrounding the injection point was analyzed with Hoechst staining and Image-ProPlus software. ** P

    Techniques Used: Over Expression, Injection, Double Staining, Labeling, Expressing, Fluorescence, Staining, Software

    17) Product Images from "Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey"

    Article Title: Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhn078

    Comparison of labeling for Mash1 ( A , B , E ) and GAD65 ( C , D , F ) in the proliferative zones of GE ( A , C ), septum ( B , D ) and dorsal telencephalon ( E , F ) in coronal sections of monkey fetal telencephalon at E47 ( E , F ) and E55 ( A – D ). Figures correspond to magnified regions indicated by boxes in Figure 1 A , C , D . ( A ) E55. In the GE many cells were labeled for Mash1 in the VZ and the SVZ, the intensity of labeling being much higher in the VZ than in the SVZ. Moreover, the cells highly labeled for Mash1 in the VZ of the ventral part of GE were organized in two bands in contrast to the dorsal part where the highly labeled cells for Mash1 were distributed in a homogeneous band (see Fig. 1 C ). ( B ) E55. In the prospective septal region (Sept) numerous cells highly labeled for Mash1 were observed in the VZ whereas the cells in the SVZ displayed a moderate level of labeling. ( C ) E55. Many cells labeled for GAD65 were observed in the GE. These cells were mainly located in the SVZ, only a few cells being present in the VZ. ( D ) E55. Many cells labeled for GAD65 were present in the SVZ of the septal region. Box: magnified region. ( E ) E47. Cresyl violet labeled section, in which the different cellular zones of the most dorsal part of the dorsal telencephalon are delineated. ( F ) E47. Some cells moderately labeled for Mash1 were observed in the proliferative zones of the most dorsal part of the dorsal telencephalon on the roof of the lateral ventricle. Most of these Mash1 positive cells were present in the upper part of the SVZ, on the border to the IZ. Some cells were also observed in the deepest part of the VZ. ( G ) E47. In contrast to Mash1 no GAD65-containing cells were observed in the VZ and SVZ of the most dorsal part of the dorsal telencephalon. However in this region, some GAD65-containing cells with a characteristic morphology of early postmitotic migrating neurons, including a leading process, and tangentially oriented, were observed in the IZ and the CP (arrows). Scale bars = 50 μm ( A – G ), 10 μm (box in D ).
    Figure Legend Snippet: Comparison of labeling for Mash1 ( A , B , E ) and GAD65 ( C , D , F ) in the proliferative zones of GE ( A , C ), septum ( B , D ) and dorsal telencephalon ( E , F ) in coronal sections of monkey fetal telencephalon at E47 ( E , F ) and E55 ( A – D ). Figures correspond to magnified regions indicated by boxes in Figure 1 A , C , D . ( A ) E55. In the GE many cells were labeled for Mash1 in the VZ and the SVZ, the intensity of labeling being much higher in the VZ than in the SVZ. Moreover, the cells highly labeled for Mash1 in the VZ of the ventral part of GE were organized in two bands in contrast to the dorsal part where the highly labeled cells for Mash1 were distributed in a homogeneous band (see Fig. 1 C ). ( B ) E55. In the prospective septal region (Sept) numerous cells highly labeled for Mash1 were observed in the VZ whereas the cells in the SVZ displayed a moderate level of labeling. ( C ) E55. Many cells labeled for GAD65 were observed in the GE. These cells were mainly located in the SVZ, only a few cells being present in the VZ. ( D ) E55. Many cells labeled for GAD65 were present in the SVZ of the septal region. Box: magnified region. ( E ) E47. Cresyl violet labeled section, in which the different cellular zones of the most dorsal part of the dorsal telencephalon are delineated. ( F ) E47. Some cells moderately labeled for Mash1 were observed in the proliferative zones of the most dorsal part of the dorsal telencephalon on the roof of the lateral ventricle. Most of these Mash1 positive cells were present in the upper part of the SVZ, on the border to the IZ. Some cells were also observed in the deepest part of the VZ. ( G ) E47. In contrast to Mash1 no GAD65-containing cells were observed in the VZ and SVZ of the most dorsal part of the dorsal telencephalon. However in this region, some GAD65-containing cells with a characteristic morphology of early postmitotic migrating neurons, including a leading process, and tangentially oriented, were observed in the IZ and the CP (arrows). Scale bars = 50 μm ( A – G ), 10 μm (box in D ).

    Techniques Used: Labeling

    Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).
    Figure Legend Snippet: Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).

    Techniques Used: Labeling, Immunohistochemistry

    Labeling patterns for Mash1 and GAD65 in coronal sections at E68, E75 and E88. (A–D) Mash1. ( A ) At E75, presence of Mash1 immunoreactivity in GE as well as in ventricular (VZ) and subventricular (SVZ) zone of the entire dorsal telencephalon. In both areas, the intensity of labeling for Mash1 decreased from the VZ to the SVZ. Few cells very lightly labeled for Mash1 were observed in the lower part of the IZ. ( B ) At E68, the same pattern of labeling for Mash1 is already present in the VZ and SVZ of the dorsal telencephalon. No labeling is observed in the upper part of IZ, the SP and the CP. ( C ) E75 Higher magnification of panel A. The numerous Mash1 highly labeled cells located in the VZ and SVZ of the dorsal telencephalon are round-shaped. ( D ) At E88, end of neurogenesis, the intensity of labeling for Mash1 was strongly decreased as compared to E68 ( B ) and E75 ( A and C ). (E–H) GAD65. ( E ) At E75, GAD65 immunolabeling was present in the GE, but also in the proliferative VZ and SVZ of the dorsal telencephalon in contrast to earlier stages ( Figs 1 D , 3 B ). The stream of migrating GAD65-containing neurons observed at E55 ( Fig. 3 B ) in the lower part of IZ and upper part of SVZ was still present (arrows). ( F ) At E68, similar labeling was already observed. In contrast to previous stages (E47–E55 compare with Figs 2 F and 3 B ) many migrating-like postmitotic neurons with one leading process were now observed in the VZ and SVZ of the dorsal telencephalon. ( G ) At E75, in addition to migrating-like postmitotic neurons, many round-shaped cells without any process were labeled for GAD65 (arrowheads). These cells displayed a similar morphology to that observed in the GE and septal region (compare with Fig. 2 C , D , and box in Fig. 2 D ). ( H ) At E75, many migrating-like GAD65 neurons in the SVZ of the dorsal telencephalon displayed multidirectional orientations. Scale bar = 550 μm ( A , E ), 250 μm ( B ), 60 μm ( C , D ) and 30 μm ( F – H ).
    Figure Legend Snippet: Labeling patterns for Mash1 and GAD65 in coronal sections at E68, E75 and E88. (A–D) Mash1. ( A ) At E75, presence of Mash1 immunoreactivity in GE as well as in ventricular (VZ) and subventricular (SVZ) zone of the entire dorsal telencephalon. In both areas, the intensity of labeling for Mash1 decreased from the VZ to the SVZ. Few cells very lightly labeled for Mash1 were observed in the lower part of the IZ. ( B ) At E68, the same pattern of labeling for Mash1 is already present in the VZ and SVZ of the dorsal telencephalon. No labeling is observed in the upper part of IZ, the SP and the CP. ( C ) E75 Higher magnification of panel A. The numerous Mash1 highly labeled cells located in the VZ and SVZ of the dorsal telencephalon are round-shaped. ( D ) At E88, end of neurogenesis, the intensity of labeling for Mash1 was strongly decreased as compared to E68 ( B ) and E75 ( A and C ). (E–H) GAD65. ( E ) At E75, GAD65 immunolabeling was present in the GE, but also in the proliferative VZ and SVZ of the dorsal telencephalon in contrast to earlier stages ( Figs 1 D , 3 B ). The stream of migrating GAD65-containing neurons observed at E55 ( Fig. 3 B ) in the lower part of IZ and upper part of SVZ was still present (arrows). ( F ) At E68, similar labeling was already observed. In contrast to previous stages (E47–E55 compare with Figs 2 F and 3 B ) many migrating-like postmitotic neurons with one leading process were now observed in the VZ and SVZ of the dorsal telencephalon. ( G ) At E75, in addition to migrating-like postmitotic neurons, many round-shaped cells without any process were labeled for GAD65 (arrowheads). These cells displayed a similar morphology to that observed in the GE and septal region (compare with Fig. 2 C , D , and box in Fig. 2 D ). ( H ) At E75, many migrating-like GAD65 neurons in the SVZ of the dorsal telencephalon displayed multidirectional orientations. Scale bar = 550 μm ( A , E ), 250 μm ( B ), 60 μm ( C , D ) and 30 μm ( F – H ).

    Techniques Used: Labeling, Immunolabeling

    Mash1 ( A , C ) and GAD65 ( D ) labeling patterns in coronal sections of monkey fetal telencephalon at E47 ( A ) and E55 ( B – D ). (A) At E47 Mash1 was highly expressed in the proliferative zone of the ventral telencephalon, the GE. Mash1 immunoreactivity was also observed in the proliferative zones of the dorsal telencephalon and followed a clear dorso-ventral decreasing gradient in the labeling intensity within the entire cortical wall. Whereas moderate level of labeling was observed in the most dorsal part of the telencephalon, on the roof of the lateral ventricle (vent) (indicated by square and magnified in Fig. 2 E ), the intensity of staining decreased in the lateral and medial wall. No Mash1 immunoreactivity was observed close to the GE, nor in the most ventral part of the medial telencephalic wall that corresponds at this level to dorsal hippocampus (dhipp). No Mash1 immunoreactivity could be seen in the thalamic nuclei (th). ( B – D ) Adjacent sections from E55 fetal brain at a rostral level in which the lateral ventricle displayed its rostral dilatation (paleocortical ventricle indicated by a star). ( B ) Cresyl violet labeled section, in which the following regions were specified: corpus callosum (CC), septum (sept), caudate nucleus (caud), putamen (put), capsula interna (CI), and primordium of the dorsal hippocampus (dhipp). ( C ) Strong intensity of labeling for Mash1 was present in the GE (box magnified in Fig. 2 A ) and septal proliferative zone (box magnified in Fig. 2 B ). ( D ) GAD65-immunolabeling was observed in the same regions of the ventral telencephalon as Mash1, the GE and the septal eminence (boxes enlarged in Fig. 2 C , D ). The levels of sections ( A , B , C , D ) are indicated by dotted lines on the schematic drawing of monkey fetal brain. Crossed arrows indicated the orientation of sections ( A , B , C , D ). Scale bar = 1 mm (same for all the figures).
    Figure Legend Snippet: Mash1 ( A , C ) and GAD65 ( D ) labeling patterns in coronal sections of monkey fetal telencephalon at E47 ( A ) and E55 ( B – D ). (A) At E47 Mash1 was highly expressed in the proliferative zone of the ventral telencephalon, the GE. Mash1 immunoreactivity was also observed in the proliferative zones of the dorsal telencephalon and followed a clear dorso-ventral decreasing gradient in the labeling intensity within the entire cortical wall. Whereas moderate level of labeling was observed in the most dorsal part of the telencephalon, on the roof of the lateral ventricle (vent) (indicated by square and magnified in Fig. 2 E ), the intensity of staining decreased in the lateral and medial wall. No Mash1 immunoreactivity was observed close to the GE, nor in the most ventral part of the medial telencephalic wall that corresponds at this level to dorsal hippocampus (dhipp). No Mash1 immunoreactivity could be seen in the thalamic nuclei (th). ( B – D ) Adjacent sections from E55 fetal brain at a rostral level in which the lateral ventricle displayed its rostral dilatation (paleocortical ventricle indicated by a star). ( B ) Cresyl violet labeled section, in which the following regions were specified: corpus callosum (CC), septum (sept), caudate nucleus (caud), putamen (put), capsula interna (CI), and primordium of the dorsal hippocampus (dhipp). ( C ) Strong intensity of labeling for Mash1 was present in the GE (box magnified in Fig. 2 A ) and septal proliferative zone (box magnified in Fig. 2 B ). ( D ) GAD65-immunolabeling was observed in the same regions of the ventral telencephalon as Mash1, the GE and the septal eminence (boxes enlarged in Fig. 2 C , D ). The levels of sections ( A , B , C , D ) are indicated by dotted lines on the schematic drawing of monkey fetal brain. Crossed arrows indicated the orientation of sections ( A , B , C , D ). Scale bar = 1 mm (same for all the figures).

    Techniques Used: Labeling, Staining, Immunolabeling

    Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 and E75 double-labeled ( A – D ) for Mash1 (red, nuclear) and GAD65 (green, cytoplasmic) or double-labeled ( E – I ) for GAD65 (green, cytoplasmic) and Ki67 (red, nuclear). ( A ) At E55, almost all Mash1-labeled cells in SVZ of the GE showed cytoplasmic labeling for GAD65, whereas none of the migrating GAD65 neurons in the IZ displayed detectable level of Mash1. ( B ) At E75, in the SVZ of GE many Mash1-containing cells were labeled for GAD65. ( C ) At E75, in the dorsal telencephalon, most Mash1-labeled cells of SVZ also contained GAD65, whereas many Mash1-labeled cells in the VZ did not. ( D ) At E75, aggregates of cells double-labeled for Mash1 and GAD65 were frequently observed in the SVZ of the dorsal telencephalon, suggesting cells ongoing division. ( E ) At E55, in the SVZ of the GE many GAD65-labeled cells were labeled for Ki67 and therefore corresponded to proliferating cells. In contrast none of the migrating GAD65-containing cells in the IZ displayed detectable level of Ki67 as illustrated at higher magnification (inset E′) and therefore likely corresponded to early postmitotic neurons. ( F , G ) At E55 in the SVZ of GE ( F ) like at E75 in the SVZ of the dorsal telencephalon ( G ) most GAD65-containing cells were labeled for Ki67 and therefore likely corresponded to progenitors of GABAergic neurons. ( H , I ) At E75, many migrating like GAD65-containing cells in the IZ ( H ) and SP ( I ) were not labeled for Ki67. Scale bars = 50 μm ( A , C , E , F , G , I ), 30 μm ( B , E ′) and 10 μm ( D , H ).
    Figure Legend Snippet: Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 and E75 double-labeled ( A – D ) for Mash1 (red, nuclear) and GAD65 (green, cytoplasmic) or double-labeled ( E – I ) for GAD65 (green, cytoplasmic) and Ki67 (red, nuclear). ( A ) At E55, almost all Mash1-labeled cells in SVZ of the GE showed cytoplasmic labeling for GAD65, whereas none of the migrating GAD65 neurons in the IZ displayed detectable level of Mash1. ( B ) At E75, in the SVZ of GE many Mash1-containing cells were labeled for GAD65. ( C ) At E75, in the dorsal telencephalon, most Mash1-labeled cells of SVZ also contained GAD65, whereas many Mash1-labeled cells in the VZ did not. ( D ) At E75, aggregates of cells double-labeled for Mash1 and GAD65 were frequently observed in the SVZ of the dorsal telencephalon, suggesting cells ongoing division. ( E ) At E55, in the SVZ of the GE many GAD65-labeled cells were labeled for Ki67 and therefore corresponded to proliferating cells. In contrast none of the migrating GAD65-containing cells in the IZ displayed detectable level of Ki67 as illustrated at higher magnification (inset E′) and therefore likely corresponded to early postmitotic neurons. ( F , G ) At E55 in the SVZ of GE ( F ) like at E75 in the SVZ of the dorsal telencephalon ( G ) most GAD65-containing cells were labeled for Ki67 and therefore likely corresponded to progenitors of GABAergic neurons. ( H , I ) At E75, many migrating like GAD65-containing cells in the IZ ( H ) and SP ( I ) were not labeled for Ki67. Scale bars = 50 μm ( A , C , E , F , G , I ), 30 μm ( B , E ′) and 10 μm ( D , H ).

    Techniques Used: Labeling

    Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 ( A – D ) and E75 ( E – G ) double-labeled for Mash1 (red, nuclear) and PGDFR-α (green, cytoplasmic). ( A ) Few cells double-labeled for PGDFR-α and Mash1 were present among the numerous cells only labeled for Mash1 observed in the SVZ of the GE. ( B , C , D ) Some PDGFR-α cells were observed in IZ and SP of the dorsal telencephalon, these cells contained very low or no detectable level of Mash1 in contrast to PDGFR-α in the GE (compare with A ). ( E ) At E75 many cells double-labeled for PDGFR-α and Mash1 were now observed in the SVZ and IZ of the dorsal telencephalon. ( F , G ) In the SP ( F ) and CP ( G ) presumed PDGFR-α containing oligodendrocytes did not show detectable level of Mash1. Scale bars = 10 μm ( A , C , D , F , G ), and 50 μm ( B , E ).
    Figure Legend Snippet: Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 ( A – D ) and E75 ( E – G ) double-labeled for Mash1 (red, nuclear) and PGDFR-α (green, cytoplasmic). ( A ) Few cells double-labeled for PGDFR-α and Mash1 were present among the numerous cells only labeled for Mash1 observed in the SVZ of the GE. ( B , C , D ) Some PDGFR-α cells were observed in IZ and SP of the dorsal telencephalon, these cells contained very low or no detectable level of Mash1 in contrast to PDGFR-α in the GE (compare with A ). ( E ) At E75 many cells double-labeled for PDGFR-α and Mash1 were now observed in the SVZ and IZ of the dorsal telencephalon. ( F , G ) In the SP ( F ) and CP ( G ) presumed PDGFR-α containing oligodendrocytes did not show detectable level of Mash1. Scale bars = 10 μm ( A , C , D , F , G ), and 50 μm ( B , E ).

    Techniques Used: Labeling

    18) Product Images from "Decreased Coenzyme Q10 Levels in Multiple System Atrophy Cerebellum"

    Article Title: Decreased Coenzyme Q10 Levels in Multiple System Atrophy Cerebellum

    Journal: Journal of Neuropathology and Experimental Neurology

    doi: 10.1093/jnen/nlw037

    Oxidative stress in cerebellum. (a) Representative images of immunohistochemistry with anti-4-HNE and anti-8-OHdG (indices of lipid and DNA oxidation, respectively) in samples from controls (CTRL) (n = 4 per staining) and multiple system atrophy (MSA) patients (n = 6 per staining). Figures are representative of 5 different areas per slide. Scale bar: 50 µm. (b) Image J analysis showed increased signals in brains of MSA cases versus controls for HNE but not for 8-OHdG. 4-HNE, 4-hydroxynonenal; 8-OHdG, 8-hydroxy-2’-deoxyguanosine; P, Purkinje cells. *p
    Figure Legend Snippet: Oxidative stress in cerebellum. (a) Representative images of immunohistochemistry with anti-4-HNE and anti-8-OHdG (indices of lipid and DNA oxidation, respectively) in samples from controls (CTRL) (n = 4 per staining) and multiple system atrophy (MSA) patients (n = 6 per staining). Figures are representative of 5 different areas per slide. Scale bar: 50 µm. (b) Image J analysis showed increased signals in brains of MSA cases versus controls for HNE but not for 8-OHdG. 4-HNE, 4-hydroxynonenal; 8-OHdG, 8-hydroxy-2’-deoxyguanosine; P, Purkinje cells. *p

    Techniques Used: Immunohistochemistry, Staining

    19) Product Images from "Endothelial damage, vascular bagging and remodeling of the microvascular bed in human microangiopathy with deep white matter lesions"

    Article Title: Endothelial damage, vascular bagging and remodeling of the microvascular bed in human microangiopathy with deep white matter lesions

    Journal: Acta Neuropathologica Communications

    doi: 10.1186/s40478-018-0632-z

    Double- and triple-label immunohistochemistry (IHC) for UEA-l, COLL4, the plasma proteins fibrinogen (FIBR) or immunoglobulin G fraction (IgG) and the endothelial marker alkaline phosphatase protein (ALPL) in 7 μm thick paraffin sections. a - b The healthy capillary in the control white matter (Co) of a NoSVD case does not have vascular bags ( a ). Vascular bag (short thick arrow) in a SVD case with multiple layers of COLL4-positive membranes around the UEA-l-labeled endothelium (white arrowhead) of a small vessel (
    Figure Legend Snippet: Double- and triple-label immunohistochemistry (IHC) for UEA-l, COLL4, the plasma proteins fibrinogen (FIBR) or immunoglobulin G fraction (IgG) and the endothelial marker alkaline phosphatase protein (ALPL) in 7 μm thick paraffin sections. a - b The healthy capillary in the control white matter (Co) of a NoSVD case does not have vascular bags ( a ). Vascular bag (short thick arrow) in a SVD case with multiple layers of COLL4-positive membranes around the UEA-l-labeled endothelium (white arrowhead) of a small vessel (

    Techniques Used: Immunohistochemistry, Marker, Labeling

    20) Product Images from "Endocytosis of a Glycosylphosphatidylinositol-anchored Protein via Clathrin-coated Vesicles, Sorting by Default in Endosomes, and Exocytosis via RAB11-positive Carriers V⃞"

    Article Title: Endocytosis of a Glycosylphosphatidylinositol-anchored Protein via Clathrin-coated Vesicles, Sorting by Default in Endosomes, and Exocytosis via RAB11-positive Carriers V⃞

    Journal: Molecular Biology of the Cell

    doi: 10.1091/mbc.E02-10-0640

    Endocytosis of VSG by clathrin-coated vesicles. (A) Section of a high-pressure frozen, freeze-substituted and Epon-embedded cell showing the FP and flagellum (FL). A clathrin-coated pit (coat marked by arrow heads) pinches off the flagellar pocket membrane and two endocytic vesicles (arrows), which have already lost their clathrin coat, can be seen in the cytoplasm. The VSG surface coat (SC) lines the flagellar pocket membrane and the inner face of coated pits and endocytic vesicles. (B and C) Cryosections labeled with anti-clathrin antibodies and PAG-6. Clathrin covers coated pits (CCP in B) and vesicles (CCV in C); a new coated pit forms at the flagellar pocket membrane in C (arrows). (D) Epon-section prepared as for A was labeled with anti-VSG antibodies and PAG-15. VSG enters a coated pit (arrow heads). (E) Cryosection labeled with rabbit anti-biotin antibodies and PAG-6. Cells were biotinylated at 0°C and then warmed to 26°C for 5 s to initiate endocytosis. Biotinylated VSG (VSG biot. ) enters a clathrin-coated pit and is present in a coated vesicle but the EXC is not labeled. The inset shows a coated pit double labeled for VSG with biotinylated anti-VSG antibodies (PAG-18) and clathrin (PAG-6; for further details, see MATERIALS AND METHODS). Bars (A–D), 250 nm; E, 125 nm.
    Figure Legend Snippet: Endocytosis of VSG by clathrin-coated vesicles. (A) Section of a high-pressure frozen, freeze-substituted and Epon-embedded cell showing the FP and flagellum (FL). A clathrin-coated pit (coat marked by arrow heads) pinches off the flagellar pocket membrane and two endocytic vesicles (arrows), which have already lost their clathrin coat, can be seen in the cytoplasm. The VSG surface coat (SC) lines the flagellar pocket membrane and the inner face of coated pits and endocytic vesicles. (B and C) Cryosections labeled with anti-clathrin antibodies and PAG-6. Clathrin covers coated pits (CCP in B) and vesicles (CCV in C); a new coated pit forms at the flagellar pocket membrane in C (arrows). (D) Epon-section prepared as for A was labeled with anti-VSG antibodies and PAG-15. VSG enters a coated pit (arrow heads). (E) Cryosection labeled with rabbit anti-biotin antibodies and PAG-6. Cells were biotinylated at 0°C and then warmed to 26°C for 5 s to initiate endocytosis. Biotinylated VSG (VSG biot. ) enters a clathrin-coated pit and is present in a coated vesicle but the EXC is not labeled. The inset shows a coated pit double labeled for VSG with biotinylated anti-VSG antibodies (PAG-18) and clathrin (PAG-6; for further details, see MATERIALS AND METHODS). Bars (A–D), 250 nm; E, 125 nm.

    Techniques Used: Labeling

    21) Product Images from "Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis"

    Article Title: Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/s13075-016-0984-3

    Id1 is expressed in ECs and ST fibroblasts. a mRNA was isolated from HMVECs and fibroblasts were isolated from NL and RA ST. mRNA was reverse transcribed into cDNA and underwent PCR for 40 cycles. RA fibroblasts showed significantly elevated Id1 expression compared to NL ST fibroblasts and HMVECs. b Id1 is expressed in RA STs. IHC was performed on RA, OA, and NL ST cryosections. Tissues were blocked and then incubated using a mouse anti-human Id1 (Abcam) primary antibody. After washing, tissues were incubated with a biotinylated anti-mouse secondary antibody (Vector Laboratories). Tissues were washed and subsequently developed with the Vectastain ABC kit (Vector Laboratories). Id1 is found on synovial cells (SNC) in the RA ST. c For immunofluorescence staining, RA, osteoarthritis (OA), and normal (NL) ST sections were fixed in cold acetone for 30 min. The STs were blocked with 5 % donkey serum and 20 % fetal bovine serum (FBS) in PBS at 37 °C for 1 h, and then incubated with rabbit anti-human Id1 antibody (Abcam, 10 μg/ml) or purified nonspecific rabbit IgG for 1 h at 37 °C in blocking buffer. The synovial tissues samples were washed with PBS, and a 1:200 dilution in blocking buffer of fluorescent-conjugated donkey anti-rabbit antibody was added and incubated for an additional 1 h at 37 °C. As shown, we could validate Id1 staining in RA ST similar to what was found using IHC (×400). FLS fibroblast-like synoviocytes, HMVEC human dermal microvascular endothelial cell, Id1 inhibitor of DNA binding 1, IgG immunoglobulin, NL normal, RA rheumatoid arthritis, SNC synovial cell
    Figure Legend Snippet: Id1 is expressed in ECs and ST fibroblasts. a mRNA was isolated from HMVECs and fibroblasts were isolated from NL and RA ST. mRNA was reverse transcribed into cDNA and underwent PCR for 40 cycles. RA fibroblasts showed significantly elevated Id1 expression compared to NL ST fibroblasts and HMVECs. b Id1 is expressed in RA STs. IHC was performed on RA, OA, and NL ST cryosections. Tissues were blocked and then incubated using a mouse anti-human Id1 (Abcam) primary antibody. After washing, tissues were incubated with a biotinylated anti-mouse secondary antibody (Vector Laboratories). Tissues were washed and subsequently developed with the Vectastain ABC kit (Vector Laboratories). Id1 is found on synovial cells (SNC) in the RA ST. c For immunofluorescence staining, RA, osteoarthritis (OA), and normal (NL) ST sections were fixed in cold acetone for 30 min. The STs were blocked with 5 % donkey serum and 20 % fetal bovine serum (FBS) in PBS at 37 °C for 1 h, and then incubated with rabbit anti-human Id1 antibody (Abcam, 10 μg/ml) or purified nonspecific rabbit IgG for 1 h at 37 °C in blocking buffer. The synovial tissues samples were washed with PBS, and a 1:200 dilution in blocking buffer of fluorescent-conjugated donkey anti-rabbit antibody was added and incubated for an additional 1 h at 37 °C. As shown, we could validate Id1 staining in RA ST similar to what was found using IHC (×400). FLS fibroblast-like synoviocytes, HMVEC human dermal microvascular endothelial cell, Id1 inhibitor of DNA binding 1, IgG immunoglobulin, NL normal, RA rheumatoid arthritis, SNC synovial cell

    Techniques Used: Isolation, Polymerase Chain Reaction, Expressing, Immunohistochemistry, Incubation, Plasmid Preparation, Immunofluorescence, Staining, Purification, Blocking Assay, Binding Assay

    SNC Id1 expression is significantly higher in inflamed ST and in the ankles of K/BxN serum-induced mice. a Id1 expression was visualized on SNCs in ST by IHC. A significantly greater percentage of SNCs were positive for Id1 in RA compared to OA and NL ST. b Percentages of cells expressing Id1 were also evaluated on SNCs from joint tissues taken from K/BxN serum-induced mice. A significantly greater percentage of SNCs positive for Id1 compared to Wt (noninduced mice) was found. c Murine tissues were blocked and then incubated using a rabbit anti-mouse Id1 antibody (CalBioreagents). After washing, tissues were incubated with a biotinylated anti-rabbit secondary antibody (Vector Laboratories). Tissues were washed and subsequently developed with the Vectastain ABC kit (Vector Laboratories). Id1 is found on SNCs near the bone in the K/BxN mouse ankles. Id1 inhibitor of DNA binding 1, IgG immunoglobulin, NL normal, OA osteoarthritis, RA rheumatoid arthritis, SNC synovial cell, ST synovial tissue, Wt wildtype
    Figure Legend Snippet: SNC Id1 expression is significantly higher in inflamed ST and in the ankles of K/BxN serum-induced mice. a Id1 expression was visualized on SNCs in ST by IHC. A significantly greater percentage of SNCs were positive for Id1 in RA compared to OA and NL ST. b Percentages of cells expressing Id1 were also evaluated on SNCs from joint tissues taken from K/BxN serum-induced mice. A significantly greater percentage of SNCs positive for Id1 compared to Wt (noninduced mice) was found. c Murine tissues were blocked and then incubated using a rabbit anti-mouse Id1 antibody (CalBioreagents). After washing, tissues were incubated with a biotinylated anti-rabbit secondary antibody (Vector Laboratories). Tissues were washed and subsequently developed with the Vectastain ABC kit (Vector Laboratories). Id1 is found on SNCs near the bone in the K/BxN mouse ankles. Id1 inhibitor of DNA binding 1, IgG immunoglobulin, NL normal, OA osteoarthritis, RA rheumatoid arthritis, SNC synovial cell, ST synovial tissue, Wt wildtype

    Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Incubation, Plasmid Preparation, Binding Assay

    22) Product Images from "N-Glycosylation of ss4 Integrin Controls the Adhesion and Motility of Keratinocytes"

    Article Title: N-Glycosylation of ss4 Integrin Controls the Adhesion and Motility of Keratinocytes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0027084

    Characterization of N -glycosylation-defective ß4 integrin-expressing keratinocytes. (A) Schematic diagram of the potential N -glycosylation sites (N X (S/T)) on the ß4 integrin subunit. The sites corresponding to the putative N -glycosylation sites on the ß4 integrin subunit (Asn 327 , Asn 491 , Asn 579 , Asn 617 , and Asn 695 ) are shown by flags. Numbers and boxes indicate the number of amino acid residue and the four intracellular fibronectin type III repeats, respectively. TM, transmembrane region. (B) Cell lysates from normal keratinocytes were immunoprecipitated using a control IgG or an anti-ß4 integrin Ab. Immunoprecipitates were run on a 6% SDS-polyacrylamide gel and probed with the biotinylated L 4 -PHA lectin (upper left panel) and E 4 -PHA lectin (upper right panel) or anti-ß4 and anti-α6 integrin Abs (lower panels). IB, immunoblot. The black and white arrowheads indicate the ß4 integrin and α6 integrin subunits, respectively. Ordinates indicate molecular sizes in kDa of marker proteins. (C) Cell morphology of the WT and ΔNß4 integrin-expressing keratinocytes during cell culture. (D) The cell-spreading area in A was calculated using computer software (Image J). Each bar represents the mean ± S.D. of triplicate assays. *p
    Figure Legend Snippet: Characterization of N -glycosylation-defective ß4 integrin-expressing keratinocytes. (A) Schematic diagram of the potential N -glycosylation sites (N X (S/T)) on the ß4 integrin subunit. The sites corresponding to the putative N -glycosylation sites on the ß4 integrin subunit (Asn 327 , Asn 491 , Asn 579 , Asn 617 , and Asn 695 ) are shown by flags. Numbers and boxes indicate the number of amino acid residue and the four intracellular fibronectin type III repeats, respectively. TM, transmembrane region. (B) Cell lysates from normal keratinocytes were immunoprecipitated using a control IgG or an anti-ß4 integrin Ab. Immunoprecipitates were run on a 6% SDS-polyacrylamide gel and probed with the biotinylated L 4 -PHA lectin (upper left panel) and E 4 -PHA lectin (upper right panel) or anti-ß4 and anti-α6 integrin Abs (lower panels). IB, immunoblot. The black and white arrowheads indicate the ß4 integrin and α6 integrin subunits, respectively. Ordinates indicate molecular sizes in kDa of marker proteins. (C) Cell morphology of the WT and ΔNß4 integrin-expressing keratinocytes during cell culture. (D) The cell-spreading area in A was calculated using computer software (Image J). Each bar represents the mean ± S.D. of triplicate assays. *p

    Techniques Used: Expressing, Immunoprecipitation, Marker, Cell Culture, Software

    23) Product Images from "Green algae Chlamydomonas reinhardtii possess endogenous sialylated N-glycans"

    Article Title: Green algae Chlamydomonas reinhardtii possess endogenous sialylated N-glycans

    Journal: FEBS Open Bio

    doi: 10.1016/j.fob.2011.10.003

    Lectin blotting analysis of C. reinhardtii proteins using MAA and SNA-I. Twenty microgram of C. reinhardtii total soluble protein (CS) and 20 μg of C. reinhardtii total membrane protein (CM) along with 2 μg fibrinogen (F), treated with sialidase (A, as indicated) and non-treated (B and C), were loaded onto the SDS–PAGE gel followed by a Western blot analysis. Sialic acid was detected using the MAA and SNA-I lectins at 50 μg/ml and 10 μg/ml, respectively, in the reaction buffer (RB) (50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM MnCl 2 ). Fibrinogen was used as a positive control. (A and B) After blocking with Carbo-free blocking buffer, the membrane was incubated with the lectins followed by avidin plus biotinylated HRP using ABC kit; (C) after blocking, the membrane was incubated with avidin plus biotinylated HRP using ABC kit. M – protein marker. Arrows show non-specific signals due to algae-derived biotin staining.
    Figure Legend Snippet: Lectin blotting analysis of C. reinhardtii proteins using MAA and SNA-I. Twenty microgram of C. reinhardtii total soluble protein (CS) and 20 μg of C. reinhardtii total membrane protein (CM) along with 2 μg fibrinogen (F), treated with sialidase (A, as indicated) and non-treated (B and C), were loaded onto the SDS–PAGE gel followed by a Western blot analysis. Sialic acid was detected using the MAA and SNA-I lectins at 50 μg/ml and 10 μg/ml, respectively, in the reaction buffer (RB) (50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM MnCl 2 ). Fibrinogen was used as a positive control. (A and B) After blocking with Carbo-free blocking buffer, the membrane was incubated with the lectins followed by avidin plus biotinylated HRP using ABC kit; (C) after blocking, the membrane was incubated with avidin plus biotinylated HRP using ABC kit. M – protein marker. Arrows show non-specific signals due to algae-derived biotin staining.

    Techniques Used: SDS Page, Western Blot, Positive Control, Blocking Assay, Incubation, Avidin-Biotin Assay, Marker, Derivative Assay, Staining

    Lectin blotting analysis of C. reinhardtii proteins using RCA 120 . Eight microgram of C. reinhardtii total soluble protein (CS), 8 μg of C. reinhardtii total membrane protein (CM), 8 μg of total soluble proteins from N. benthamiana (PC), 8 μg of total soluble proteins from N. benthamiana infiltrated with human β1,4-galactosyltransferase (PT), and 250 ng of fibrinogen (F) were loaded onto the SDS–PAGE gel followed by a Western blot analysis. Galactosylated proteins were detected using the RCA 120 lectin at 10 μg/ml in PBS with 0.05% Tween. Fibrinogen (Cat. No. F8630, Sigma, St. Louis, MO) was used as a positive control. (A) After blocking with Carbo-free blocking buffer, membranes were incubated with the 1,4-galactose residue-specific lectin followed by avidin plus biotinylated HRP using ABC kit (Vector Laboratories); (B) after blocking, the membrane was incubated with avidin plus biotinylated HRP using ABC kit. M – protein marker. Arrows show non-specific signals due to algae-derived biotin staining.
    Figure Legend Snippet: Lectin blotting analysis of C. reinhardtii proteins using RCA 120 . Eight microgram of C. reinhardtii total soluble protein (CS), 8 μg of C. reinhardtii total membrane protein (CM), 8 μg of total soluble proteins from N. benthamiana (PC), 8 μg of total soluble proteins from N. benthamiana infiltrated with human β1,4-galactosyltransferase (PT), and 250 ng of fibrinogen (F) were loaded onto the SDS–PAGE gel followed by a Western blot analysis. Galactosylated proteins were detected using the RCA 120 lectin at 10 μg/ml in PBS with 0.05% Tween. Fibrinogen (Cat. No. F8630, Sigma, St. Louis, MO) was used as a positive control. (A) After blocking with Carbo-free blocking buffer, membranes were incubated with the 1,4-galactose residue-specific lectin followed by avidin plus biotinylated HRP using ABC kit (Vector Laboratories); (B) after blocking, the membrane was incubated with avidin plus biotinylated HRP using ABC kit. M – protein marker. Arrows show non-specific signals due to algae-derived biotin staining.

    Techniques Used: SDS Page, Western Blot, Positive Control, Blocking Assay, Incubation, Avidin-Biotin Assay, Plasmid Preparation, Marker, Derivative Assay, Staining

    24) Product Images from "Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey"

    Article Title: Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhn078

    Comparison of labeling for Mash1 ( A , B , E ) and GAD65 ( C , D , F ) in the proliferative zones of GE ( A , C ), septum ( B , D ) and dorsal telencephalon ( E , F ) in coronal sections of monkey fetal telencephalon at E47 ( E , F ) and E55 ( A – D ). Figures correspond to magnified regions indicated by boxes in Figure 1 A , C , D . ( A ) E55. In the GE many cells were labeled for Mash1 in the VZ and the SVZ, the intensity of labeling being much higher in the VZ than in the SVZ. Moreover, the cells highly labeled for Mash1 in the VZ of the ventral part of GE were organized in two bands in contrast to the dorsal part where the highly labeled cells for Mash1 were distributed in a homogeneous band (see Fig. 1 C ). ( B ) E55. In the prospective septal region (Sept) numerous cells highly labeled for Mash1 were observed in the VZ whereas the cells in the SVZ displayed a moderate level of labeling. ( C ) E55. Many cells labeled for GAD65 were observed in the GE. These cells were mainly located in the SVZ, only a few cells being present in the VZ. ( D ) E55. Many cells labeled for GAD65 were present in the SVZ of the septal region. Box: magnified region. ( E ) E47. Cresyl violet labeled section, in which the different cellular zones of the most dorsal part of the dorsal telencephalon are delineated. ( F ) E47. Some cells moderately labeled for Mash1 were observed in the proliferative zones of the most dorsal part of the dorsal telencephalon on the roof of the lateral ventricle. Most of these Mash1 positive cells were present in the upper part of the SVZ, on the border to the IZ. Some cells were also observed in the deepest part of the VZ. ( G ) E47. In contrast to Mash1 no GAD65-containing cells were observed in the VZ and SVZ of the most dorsal part of the dorsal telencephalon. However in this region, some GAD65-containing cells with a characteristic morphology of early postmitotic migrating neurons, including a leading process, and tangentially oriented, were observed in the IZ and the CP (arrows). Scale bars = 50 μm ( A – G ), 10 μm (box in D ).
    Figure Legend Snippet: Comparison of labeling for Mash1 ( A , B , E ) and GAD65 ( C , D , F ) in the proliferative zones of GE ( A , C ), septum ( B , D ) and dorsal telencephalon ( E , F ) in coronal sections of monkey fetal telencephalon at E47 ( E , F ) and E55 ( A – D ). Figures correspond to magnified regions indicated by boxes in Figure 1 A , C , D . ( A ) E55. In the GE many cells were labeled for Mash1 in the VZ and the SVZ, the intensity of labeling being much higher in the VZ than in the SVZ. Moreover, the cells highly labeled for Mash1 in the VZ of the ventral part of GE were organized in two bands in contrast to the dorsal part where the highly labeled cells for Mash1 were distributed in a homogeneous band (see Fig. 1 C ). ( B ) E55. In the prospective septal region (Sept) numerous cells highly labeled for Mash1 were observed in the VZ whereas the cells in the SVZ displayed a moderate level of labeling. ( C ) E55. Many cells labeled for GAD65 were observed in the GE. These cells were mainly located in the SVZ, only a few cells being present in the VZ. ( D ) E55. Many cells labeled for GAD65 were present in the SVZ of the septal region. Box: magnified region. ( E ) E47. Cresyl violet labeled section, in which the different cellular zones of the most dorsal part of the dorsal telencephalon are delineated. ( F ) E47. Some cells moderately labeled for Mash1 were observed in the proliferative zones of the most dorsal part of the dorsal telencephalon on the roof of the lateral ventricle. Most of these Mash1 positive cells were present in the upper part of the SVZ, on the border to the IZ. Some cells were also observed in the deepest part of the VZ. ( G ) E47. In contrast to Mash1 no GAD65-containing cells were observed in the VZ and SVZ of the most dorsal part of the dorsal telencephalon. However in this region, some GAD65-containing cells with a characteristic morphology of early postmitotic migrating neurons, including a leading process, and tangentially oriented, were observed in the IZ and the CP (arrows). Scale bars = 50 μm ( A – G ), 10 μm (box in D ).

    Techniques Used: Labeling

    Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).
    Figure Legend Snippet: Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).

    Techniques Used: Labeling, Immunohistochemistry

    Labeling patterns for Mash1 and GAD65 in coronal sections at E68, E75 and E88. (A–D) Mash1. ( A ) At E75, presence of Mash1 immunoreactivity in GE as well as in ventricular (VZ) and subventricular (SVZ) zone of the entire dorsal telencephalon. In both areas, the intensity of labeling for Mash1 decreased from the VZ to the SVZ. Few cells very lightly labeled for Mash1 were observed in the lower part of the IZ. ( B ) At E68, the same pattern of labeling for Mash1 is already present in the VZ and SVZ of the dorsal telencephalon. No labeling is observed in the upper part of IZ, the SP and the CP. ( C ) E75 Higher magnification of panel A. The numerous Mash1 highly labeled cells located in the VZ and SVZ of the dorsal telencephalon are round-shaped. ( D ) At E88, end of neurogenesis, the intensity of labeling for Mash1 was strongly decreased as compared to E68 ( B ) and E75 ( A and C ). (E–H) GAD65. ( E ) At E75, GAD65 immunolabeling was present in the GE, but also in the proliferative VZ and SVZ of the dorsal telencephalon in contrast to earlier stages ( Figs 1 D , 3 B ). The stream of migrating GAD65-containing neurons observed at E55 ( Fig. 3 B ) in the lower part of IZ and upper part of SVZ was still present (arrows). ( F ) At E68, similar labeling was already observed. In contrast to previous stages (E47–E55 compare with Figs 2 F and 3 B ) many migrating-like postmitotic neurons with one leading process were now observed in the VZ and SVZ of the dorsal telencephalon. ( G ) At E75, in addition to migrating-like postmitotic neurons, many round-shaped cells without any process were labeled for GAD65 (arrowheads). These cells displayed a similar morphology to that observed in the GE and septal region (compare with Fig. 2 C , D , and box in Fig. 2 D ). ( H ) At E75, many migrating-like GAD65 neurons in the SVZ of the dorsal telencephalon displayed multidirectional orientations. Scale bar = 550 μm ( A , E ), 250 μm ( B ), 60 μm ( C , D ) and 30 μm ( F – H ).
    Figure Legend Snippet: Labeling patterns for Mash1 and GAD65 in coronal sections at E68, E75 and E88. (A–D) Mash1. ( A ) At E75, presence of Mash1 immunoreactivity in GE as well as in ventricular (VZ) and subventricular (SVZ) zone of the entire dorsal telencephalon. In both areas, the intensity of labeling for Mash1 decreased from the VZ to the SVZ. Few cells very lightly labeled for Mash1 were observed in the lower part of the IZ. ( B ) At E68, the same pattern of labeling for Mash1 is already present in the VZ and SVZ of the dorsal telencephalon. No labeling is observed in the upper part of IZ, the SP and the CP. ( C ) E75 Higher magnification of panel A. The numerous Mash1 highly labeled cells located in the VZ and SVZ of the dorsal telencephalon are round-shaped. ( D ) At E88, end of neurogenesis, the intensity of labeling for Mash1 was strongly decreased as compared to E68 ( B ) and E75 ( A and C ). (E–H) GAD65. ( E ) At E75, GAD65 immunolabeling was present in the GE, but also in the proliferative VZ and SVZ of the dorsal telencephalon in contrast to earlier stages ( Figs 1 D , 3 B ). The stream of migrating GAD65-containing neurons observed at E55 ( Fig. 3 B ) in the lower part of IZ and upper part of SVZ was still present (arrows). ( F ) At E68, similar labeling was already observed. In contrast to previous stages (E47–E55 compare with Figs 2 F and 3 B ) many migrating-like postmitotic neurons with one leading process were now observed in the VZ and SVZ of the dorsal telencephalon. ( G ) At E75, in addition to migrating-like postmitotic neurons, many round-shaped cells without any process were labeled for GAD65 (arrowheads). These cells displayed a similar morphology to that observed in the GE and septal region (compare with Fig. 2 C , D , and box in Fig. 2 D ). ( H ) At E75, many migrating-like GAD65 neurons in the SVZ of the dorsal telencephalon displayed multidirectional orientations. Scale bar = 550 μm ( A , E ), 250 μm ( B ), 60 μm ( C , D ) and 30 μm ( F – H ).

    Techniques Used: Labeling, Immunolabeling

    Mash1 ( A , C ) and GAD65 ( D ) labeling patterns in coronal sections of monkey fetal telencephalon at E47 ( A ) and E55 ( B – D ). (A) At E47 Mash1 was highly expressed in the proliferative zone of the ventral telencephalon, the GE. Mash1 immunoreactivity was also observed in the proliferative zones of the dorsal telencephalon and followed a clear dorso-ventral decreasing gradient in the labeling intensity within the entire cortical wall. Whereas moderate level of labeling was observed in the most dorsal part of the telencephalon, on the roof of the lateral ventricle (vent) (indicated by square and magnified in Fig. 2 E ), the intensity of staining decreased in the lateral and medial wall. No Mash1 immunoreactivity was observed close to the GE, nor in the most ventral part of the medial telencephalic wall that corresponds at this level to dorsal hippocampus (dhipp). No Mash1 immunoreactivity could be seen in the thalamic nuclei (th). ( B – D ) Adjacent sections from E55 fetal brain at a rostral level in which the lateral ventricle displayed its rostral dilatation (paleocortical ventricle indicated by a star). ( B ) Cresyl violet labeled section, in which the following regions were specified: corpus callosum (CC), septum (sept), caudate nucleus (caud), putamen (put), capsula interna (CI), and primordium of the dorsal hippocampus (dhipp). ( C ) Strong intensity of labeling for Mash1 was present in the GE (box magnified in Fig. 2 A ) and septal proliferative zone (box magnified in Fig. 2 B ). ( D ) GAD65-immunolabeling was observed in the same regions of the ventral telencephalon as Mash1, the GE and the septal eminence (boxes enlarged in Fig. 2 C , D ). The levels of sections ( A , B , C , D ) are indicated by dotted lines on the schematic drawing of monkey fetal brain. Crossed arrows indicated the orientation of sections ( A , B , C , D ). Scale bar = 1 mm (same for all the figures).
    Figure Legend Snippet: Mash1 ( A , C ) and GAD65 ( D ) labeling patterns in coronal sections of monkey fetal telencephalon at E47 ( A ) and E55 ( B – D ). (A) At E47 Mash1 was highly expressed in the proliferative zone of the ventral telencephalon, the GE. Mash1 immunoreactivity was also observed in the proliferative zones of the dorsal telencephalon and followed a clear dorso-ventral decreasing gradient in the labeling intensity within the entire cortical wall. Whereas moderate level of labeling was observed in the most dorsal part of the telencephalon, on the roof of the lateral ventricle (vent) (indicated by square and magnified in Fig. 2 E ), the intensity of staining decreased in the lateral and medial wall. No Mash1 immunoreactivity was observed close to the GE, nor in the most ventral part of the medial telencephalic wall that corresponds at this level to dorsal hippocampus (dhipp). No Mash1 immunoreactivity could be seen in the thalamic nuclei (th). ( B – D ) Adjacent sections from E55 fetal brain at a rostral level in which the lateral ventricle displayed its rostral dilatation (paleocortical ventricle indicated by a star). ( B ) Cresyl violet labeled section, in which the following regions were specified: corpus callosum (CC), septum (sept), caudate nucleus (caud), putamen (put), capsula interna (CI), and primordium of the dorsal hippocampus (dhipp). ( C ) Strong intensity of labeling for Mash1 was present in the GE (box magnified in Fig. 2 A ) and septal proliferative zone (box magnified in Fig. 2 B ). ( D ) GAD65-immunolabeling was observed in the same regions of the ventral telencephalon as Mash1, the GE and the septal eminence (boxes enlarged in Fig. 2 C , D ). The levels of sections ( A , B , C , D ) are indicated by dotted lines on the schematic drawing of monkey fetal brain. Crossed arrows indicated the orientation of sections ( A , B , C , D ). Scale bar = 1 mm (same for all the figures).

    Techniques Used: Labeling, Staining, Immunolabeling

    Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 and E75 double-labeled ( A – D ) for Mash1 (red, nuclear) and GAD65 (green, cytoplasmic) or double-labeled ( E – I ) for GAD65 (green, cytoplasmic) and Ki67 (red, nuclear). ( A ) At E55, almost all Mash1-labeled cells in SVZ of the GE showed cytoplasmic labeling for GAD65, whereas none of the migrating GAD65 neurons in the IZ displayed detectable level of Mash1. ( B ) At E75, in the SVZ of GE many Mash1-containing cells were labeled for GAD65. ( C ) At E75, in the dorsal telencephalon, most Mash1-labeled cells of SVZ also contained GAD65, whereas many Mash1-labeled cells in the VZ did not. ( D ) At E75, aggregates of cells double-labeled for Mash1 and GAD65 were frequently observed in the SVZ of the dorsal telencephalon, suggesting cells ongoing division. ( E ) At E55, in the SVZ of the GE many GAD65-labeled cells were labeled for Ki67 and therefore corresponded to proliferating cells. In contrast none of the migrating GAD65-containing cells in the IZ displayed detectable level of Ki67 as illustrated at higher magnification (inset E′) and therefore likely corresponded to early postmitotic neurons. ( F , G ) At E55 in the SVZ of GE ( F ) like at E75 in the SVZ of the dorsal telencephalon ( G ) most GAD65-containing cells were labeled for Ki67 and therefore likely corresponded to progenitors of GABAergic neurons. ( H , I ) At E75, many migrating like GAD65-containing cells in the IZ ( H ) and SP ( I ) were not labeled for Ki67. Scale bars = 50 μm ( A , C , E , F , G , I ), 30 μm ( B , E ′) and 10 μm ( D , H ).
    Figure Legend Snippet: Confocal micrographs of coronal sections of monkey fetal telencephalon at E55 and E75 double-labeled ( A – D ) for Mash1 (red, nuclear) and GAD65 (green, cytoplasmic) or double-labeled ( E – I ) for GAD65 (green, cytoplasmic) and Ki67 (red, nuclear). ( A ) At E55, almost all Mash1-labeled cells in SVZ of the GE showed cytoplasmic labeling for GAD65, whereas none of the migrating GAD65 neurons in the IZ displayed detectable level of Mash1. ( B ) At E75, in the SVZ of GE many Mash1-containing cells were labeled for GAD65. ( C ) At E75, in the dorsal telencephalon, most Mash1-labeled cells of SVZ also contained GAD65, whereas many Mash1-labeled cells in the VZ did not. ( D ) At E75, aggregates of cells double-labeled for Mash1 and GAD65 were frequently observed in the SVZ of the dorsal telencephalon, suggesting cells ongoing division. ( E ) At E55, in the SVZ of the GE many GAD65-labeled cells were labeled for Ki67 and therefore corresponded to proliferating cells. In contrast none of the migrating GAD65-containing cells in the IZ displayed detectable level of Ki67 as illustrated at higher magnification (inset E′) and therefore likely corresponded to early postmitotic neurons. ( F , G ) At E55 in the SVZ of GE ( F ) like at E75 in the SVZ of the dorsal telencephalon ( G ) most GAD65-containing cells were labeled for Ki67 and therefore likely corresponded to progenitors of GABAergic neurons. ( H , I ) At E75, many migrating like GAD65-containing cells in the IZ ( H ) and SP ( I ) were not labeled for Ki67. Scale bars = 50 μm ( A , C , E , F , G , I ), 30 μm ( B , E ′) and 10 μm ( D , H ).

    Techniques Used: Labeling

    25) Product Images from "GM1 ganglioside-independent intoxication by Cholera toxin"

    Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006862

    CTB binds to Le X -carrying proteins in HL-60 cells. (A) Histogram from flow cytometry analysis of CTB-binding to HL-60 cells following pre-treatment of the cells with AAL (10 μg/ml) or pre-treatment of CTB with sugars (50 mM). (B) gMFI of CTB binding to HL-60 cells cultured with the indicated inhibitors (*** = p
    Figure Legend Snippet: CTB binds to Le X -carrying proteins in HL-60 cells. (A) Histogram from flow cytometry analysis of CTB-binding to HL-60 cells following pre-treatment of the cells with AAL (10 μg/ml) or pre-treatment of CTB with sugars (50 mM). (B) gMFI of CTB binding to HL-60 cells cultured with the indicated inhibitors (*** = p

    Techniques Used: CtB Assay, Flow Cytometry, Cytometry, Binding Assay, Cell Culture

    CTB binds to fucosylated structures expressed on human granulocytes. (A) Representative histograms from flow cytometry analysis of CTB- (red) and OVA- (white) binding to cell types in human peripheral blood. (B) Bar graphs show the geometric mean fluorescent index (gMFI) of CTB binding (with OVA gMFI subtracted) to human whole blood (n = 8) and murine splenocytes (n = 4–6). Each dot represents one donor/mouse. (C) gMFI of CTB and OVA binding to the different cell types from human whole blood. Each pair of dots represents one donor. (D-E) Histogram and bar graph (n = 8) showing blocking of CTB binding to human granulocytes by pre-treating CTB with (D) L-, D-fucose and D-galactose or (E) with the lectins AAL or MAL-II. (F) Histograms and bar graphs (n = 6) showing the blocking of CTB binding to murine (wt or KO) cells by either pre-treating the cells with lectins or pre-treating CTB with sugars. For panel (C) significance was calculated using paired t-test and for (D-E) one-way-ANOVA was used with Tukey correction (*** = p
    Figure Legend Snippet: CTB binds to fucosylated structures expressed on human granulocytes. (A) Representative histograms from flow cytometry analysis of CTB- (red) and OVA- (white) binding to cell types in human peripheral blood. (B) Bar graphs show the geometric mean fluorescent index (gMFI) of CTB binding (with OVA gMFI subtracted) to human whole blood (n = 8) and murine splenocytes (n = 4–6). Each dot represents one donor/mouse. (C) gMFI of CTB and OVA binding to the different cell types from human whole blood. Each pair of dots represents one donor. (D-E) Histogram and bar graph (n = 8) showing blocking of CTB binding to human granulocytes by pre-treating CTB with (D) L-, D-fucose and D-galactose or (E) with the lectins AAL or MAL-II. (F) Histograms and bar graphs (n = 6) showing the blocking of CTB binding to murine (wt or KO) cells by either pre-treating the cells with lectins or pre-treating CTB with sugars. For panel (C) significance was calculated using paired t-test and for (D-E) one-way-ANOVA was used with Tukey correction (*** = p

    Techniques Used: CtB Assay, Flow Cytometry, Cytometry, Binding Assay, Blocking Assay

    GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.
    Figure Legend Snippet: GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.

    Techniques Used: Inhibition, Cell Culture, Staining, CtB Assay, Fluorescence, Flow Cytometry, Cytometry, Luminescence Assay, Polyacrylamide Gel Electrophoresis

    26) Product Images from "Terminal Bronchioles Harbor a Unique Airway Stem Cell Population That Localizes to the Bronchoalveolar Duct Junction"

    Article Title: Terminal Bronchioles Harbor a Unique Airway Stem Cell Population That Localizes to the Bronchoalveolar Duct Junction

    Journal: The American Journal of Pathology

    doi:

    CE cells adjacent to the BADJ are pollutant-resistant and represent the initial proliferative epithelial population. Mice were exposed to 275 mg/kg of naphthalene and recovered for 0, 6, 12, 24, and 36 hours ( A to E , respectively). Proliferating mitotic cells were labeled with 2.5 mCi of [ 3 H]-TdR/kg body weight 1 hour before sacrifice. CE cells were detected by immunohistochemistry using diaminobenzidine detection (brown) and sites of [ 3 H]-TdR incorporation identified by autoradiography (black grains). Nuclei were counterstained with hematoxylin. Naphthalene-resistant CE cells are detectable adjacent to the BADJ at all time points ( A–E , arrows ), and are the initial proliferative epithelial cell type 36 hours after naphthalene administration ( E , black grains). Arrowheads in B and E indicate underlying proliferative fibroblasts. Original magnifications, ×400.
    Figure Legend Snippet: CE cells adjacent to the BADJ are pollutant-resistant and represent the initial proliferative epithelial population. Mice were exposed to 275 mg/kg of naphthalene and recovered for 0, 6, 12, 24, and 36 hours ( A to E , respectively). Proliferating mitotic cells were labeled with 2.5 mCi of [ 3 H]-TdR/kg body weight 1 hour before sacrifice. CE cells were detected by immunohistochemistry using diaminobenzidine detection (brown) and sites of [ 3 H]-TdR incorporation identified by autoradiography (black grains). Nuclei were counterstained with hematoxylin. Naphthalene-resistant CE cells are detectable adjacent to the BADJ at all time points ( A–E , arrows ), and are the initial proliferative epithelial cell type 36 hours after naphthalene administration ( E , black grains). Arrowheads in B and E indicate underlying proliferative fibroblasts. Original magnifications, ×400.

    Techniques Used: Mouse Assay, Labeling, Immunohistochemistry, Autoradiography

    27) Product Images from "GM1 ganglioside-independent intoxication by Cholera toxin"

    Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006862

    CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p
    Figure Legend Snippet: CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p

    Techniques Used: Incubation

    GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.
    Figure Legend Snippet: GM1-deficient C6 cells are sensitized to CT by inhibition of sialylation or GSL biosynthesis. (A-E) C6 cells were cultured with the indicated inhibitors for 72 h followed by: (A) Staining was then performed with biotin-CTB, followed by DTAF-streptavidin. Fluorescence was measured by flow cytometry, represented here by MFI. (B) 1 h exposure to CT after which accumulated cAMP was measured by the cAMP-Glo™ luminescence assay. Luminescence signal is inversely proportional to cAMP levels. (C) As in panel A , but stained with biotin-PNA, followed by DTAF-streptavidin (D) Cell lysates were separated by PAGE and probed with biotin-PNA, biotin-CTB, or no biotinylated reagent, followed by streptavidin-peroxidase conjugate and development with chemiluminescent substrate. Equivalent amounts of protein were loaded in each lane and blots were probed with an anti-α-tubulin or anti-GAPDH antibody to confirm equivalent loading. (E) As in panel B , but brefeldin A (BFA) was added 1 h prior to CT addition and was also present during CT induction.

    Techniques Used: Inhibition, Cell Culture, Staining, CtB Assay, Fluorescence, Flow Cytometry, Cytometry, Luminescence Assay, Polyacrylamide Gel Electrophoresis

    28) Product Images from "Marginal Zone B-Cell Depletion Impairs Murine Host Defense against Borrelia burgdorferi Infection ▿"

    Article Title: Marginal Zone B-Cell Depletion Impairs Murine Host Defense against Borrelia burgdorferi Infection ▿

    Journal:

    doi: 10.1128/IAI.00422-07

    MZB cell depletion reduces B. burgdorferi -specific IgM and IgG titers and increases pathogen burden at 7 days of infection. Ab titers are the reciprocal endpoint positive titers of B. burgdorferi -specific IgM (A) or IgG (C) in serum obtained from 7-day-infected
    Figure Legend Snippet: MZB cell depletion reduces B. burgdorferi -specific IgM and IgG titers and increases pathogen burden at 7 days of infection. Ab titers are the reciprocal endpoint positive titers of B. burgdorferi -specific IgM (A) or IgG (C) in serum obtained from 7-day-infected

    Techniques Used: Infection

    29) Product Images from "Dual role for Fc? receptors in host defense and disease in Borrelia burgdorferi-infected mice"

    Article Title: Dual role for Fc? receptors in host defense and disease in Borrelia burgdorferi-infected mice

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2014.00075

    Anti- B. burgdorferi IgG subclass titers are different in the single and double knockout mice . Anti- B. burgdorferi specific titers were determined by ELISA and are reported as the reciprocal of the endpoint positive titers. The FcεR γ −/− MyD88 −/− anti- B. burgdorferi IgG1 titers are significantly higher than the MyD88 −/− titers ( P = 0.0113). The MyD88 −/− IgG 3 titers are significantly lower than the FcεR γ −/− MyD88 −/− titers ( P = 0.0025) as well as the WT and FcεR γ −/− titers ( P = 0.01 and.0025, respectively).
    Figure Legend Snippet: Anti- B. burgdorferi IgG subclass titers are different in the single and double knockout mice . Anti- B. burgdorferi specific titers were determined by ELISA and are reported as the reciprocal of the endpoint positive titers. The FcεR γ −/− MyD88 −/− anti- B. burgdorferi IgG1 titers are significantly higher than the MyD88 −/− titers ( P = 0.0113). The MyD88 −/− IgG 3 titers are significantly lower than the FcεR γ −/− MyD88 −/− titers ( P = 0.0025) as well as the WT and FcεR γ −/− titers ( P = 0.01 and.0025, respectively).

    Techniques Used: Double Knockout, Mouse Assay, Enzyme-linked Immunosorbent Assay

    30) Product Images from "Activation of Nerve Growth Factor-Induced B? by Methylene-Substituted Diindolylmethanes in Bladder Cancer Cells Induces Apoptosis and Inhibits Tumor Growth S⃞"

    Article Title: Activation of Nerve Growth Factor-Induced B? by Methylene-Substituted Diindolylmethanes in Bladder Cancer Cells Induces Apoptosis and Inhibits Tumor Growth S⃞

    Journal: Molecular Pharmacology

    doi: 10.1124/mol.109.061143

    Nur77-active C-DIMs induce apoptosis in UC-5 and KU7 cells and also act through nuclear Nur77. A, effects of leptomycin B on PARP cleavage. UC-5 cells were treated for 24 h with DIM-C-pPhOCH 3 alone or in the presence of leptomycin B (0.05 ng/ml), and Western blot analysis of nuclear and cytosolic cell lysates was used to detect Nur77 and PARP cleavage. Sp1, loading control for nuclear fraction; N.S., nonspecific band for loading control. B, immunostaining for Nur77. UC-5 cells were treated with DMSO or 15 μM DIM-C-pPhOCH 3 for 24 h and immunostained with IgG or Nur77 antibodies. C, dose-dependent induction of apoptosis by C-DIMs. UC-5 and KU7 cells were treated with DMSO or different concentrations of DIM-C-pPhOCH 3 or DIM-C-Ph for 48 h. D, inhibition of Nur77-active C-DIMs-induced apoptosis by Z-VAD-fmk. UC-5 cells were treated with Nur77-active C-DIMs alone or in combination with 10 μM Z-VAD-fmk, and whole-cell lysates were analyzed by Western blot analysis.
    Figure Legend Snippet: Nur77-active C-DIMs induce apoptosis in UC-5 and KU7 cells and also act through nuclear Nur77. A, effects of leptomycin B on PARP cleavage. UC-5 cells were treated for 24 h with DIM-C-pPhOCH 3 alone or in the presence of leptomycin B (0.05 ng/ml), and Western blot analysis of nuclear and cytosolic cell lysates was used to detect Nur77 and PARP cleavage. Sp1, loading control for nuclear fraction; N.S., nonspecific band for loading control. B, immunostaining for Nur77. UC-5 cells were treated with DMSO or 15 μM DIM-C-pPhOCH 3 for 24 h and immunostained with IgG or Nur77 antibodies. C, dose-dependent induction of apoptosis by C-DIMs. UC-5 and KU7 cells were treated with DMSO or different concentrations of DIM-C-pPhOCH 3 or DIM-C-Ph for 48 h. D, inhibition of Nur77-active C-DIMs-induced apoptosis by Z-VAD-fmk. UC-5 cells were treated with Nur77-active C-DIMs alone or in combination with 10 μM Z-VAD-fmk, and whole-cell lysates were analyzed by Western blot analysis.

    Techniques Used: Activated Clotting Time Assay, Western Blot, Immunostaining, Inhibition

    31) Product Images from "Mitogen- and stress-activated protein kinase 1-induced neuroprotection in Huntington's disease: role on chromatin remodeling at the PGC-1-alpha promoter"

    Article Title: Mitogen- and stress-activated protein kinase 1-induced neuroprotection in Huntington's disease: role on chromatin remodeling at the PGC-1-alpha promoter

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddr148

    MSK-1-expressing cells exhibit DARPP-32 immunoreactivity in the area surrounding the injection point 10 weeks post-infection. ( A ) Rats were infected as indicated in Fig. 1 . ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels), 10 weeks later. Note the down-regulation of both MSK-1 and DARPP-32 in the right striatum infected with LV-Exp-Ht alone. Note also the preserved expression of both MSK-1 and DARPP-32 around the injection point on the left side infected with both LV-Exp-Htt and LV-MSK-1. The right panels show a higher magnification of the area surrounding the LV-Exp + MSK-1 injection point (see square box in the middle panels). Note that all three markers, Exp-Ht, MSK-1 and DARPP-32, are expressed in this area (white arrow). ( C ) The Exp-Htt volume expression in the rostro-caudal extension of the striatum and ( D ) the DARPP-32 loss/Exp-Htt infected area were measured as described in Fig. 1 . Statistics: means ± SEM; eight rats per group. ns, not significant; * P
    Figure Legend Snippet: MSK-1-expressing cells exhibit DARPP-32 immunoreactivity in the area surrounding the injection point 10 weeks post-infection. ( A ) Rats were infected as indicated in Fig. 1 . ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels), 10 weeks later. Note the down-regulation of both MSK-1 and DARPP-32 in the right striatum infected with LV-Exp-Ht alone. Note also the preserved expression of both MSK-1 and DARPP-32 around the injection point on the left side infected with both LV-Exp-Htt and LV-MSK-1. The right panels show a higher magnification of the area surrounding the LV-Exp + MSK-1 injection point (see square box in the middle panels). Note that all three markers, Exp-Ht, MSK-1 and DARPP-32, are expressed in this area (white arrow). ( C ) The Exp-Htt volume expression in the rostro-caudal extension of the striatum and ( D ) the DARPP-32 loss/Exp-Htt infected area were measured as described in Fig. 1 . Statistics: means ± SEM; eight rats per group. ns, not significant; * P

    Techniques Used: Expressing, Injection, Infection

    MSK-1 overexpression in the striatum induces histone H3 and CREB hyperphosphorylation. Rat striata were infected with LV-Htt or LV-Exp-Htt, with or without MSK-1. ( A ) Immunohistochemical detection of phospho-Ser 10 -H3 was performed 4 weeks later with an anti-rabbit Cy3-coupled secondary antibody (upper panels). MSK-1 expression was detected after co-labeling with an anti-mouse anti-FITC secondary antibody, on the same section (lower panels). Note that a majority of cells express P-H3 in the LV-Htt + MSK-1- and LV-Exp-Htt + MSK-1-infected striata but not in the striata infected with LV-Htt or LV-Exp-Htt alone. ( B ) Immunohistochemical detection of phospho-Ser 133 -CREB was performed 4 weeks post-infection. MSK-1 expression was detected as indicated in (A). Note that MSK-1 over-expression enhanced P-CREB immunoreactivity in Htt + MSK-1- and Exp-Htt + MSK-1-infected striata. ( C ) MSK-1, ( D ) P-H3- and ( E ) P-CREB-immunoreactive neurons were counted in the infected areas of the striatum, as indicated in Figure 3 . * P
    Figure Legend Snippet: MSK-1 overexpression in the striatum induces histone H3 and CREB hyperphosphorylation. Rat striata were infected with LV-Htt or LV-Exp-Htt, with or without MSK-1. ( A ) Immunohistochemical detection of phospho-Ser 10 -H3 was performed 4 weeks later with an anti-rabbit Cy3-coupled secondary antibody (upper panels). MSK-1 expression was detected after co-labeling with an anti-mouse anti-FITC secondary antibody, on the same section (lower panels). Note that a majority of cells express P-H3 in the LV-Htt + MSK-1- and LV-Exp-Htt + MSK-1-infected striata but not in the striata infected with LV-Htt or LV-Exp-Htt alone. ( B ) Immunohistochemical detection of phospho-Ser 133 -CREB was performed 4 weeks post-infection. MSK-1 expression was detected as indicated in (A). Note that MSK-1 over-expression enhanced P-CREB immunoreactivity in Htt + MSK-1- and Exp-Htt + MSK-1-infected striata. ( C ) MSK-1, ( D ) P-H3- and ( E ) P-CREB-immunoreactive neurons were counted in the infected areas of the striatum, as indicated in Figure 3 . * P

    Techniques Used: Over Expression, Infection, Immunohistochemistry, Expressing, Labeling

    Exp-Htt-induced down-regulation of DARPP-32 is attenuated by MSK-1 overexpression 4 weeks post-infection. ( A ) The left striatum was infected with a lentiviral construct containing a gene fragment of huntingtin with 82 polyglutamine repeats (Exp-Htt) and the right striatum was infected with LV-Exp-Htt and another lentiviral construct containing the MSK-1 gene (Exp-Htt + MSK-1). ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels) was performed with specific antibodies (see Materials and Methods), 4 weeks after infection. ( C ) The volume of striatal tissue showing Exp-Htt expression or DARPP-32 loss was measured on adjacent coronal sections across the rostro-caudal extension of the striatum. Seven to 10 sections per rat were analyzed (each section was 25 µm thick and was separated by 175 µm). ( D ) Graphical representation of the Exp-Htt volume expression in the rostro-caudal extension of the striatum. ( E ) Graphical representation of DARPP-32 loss/Exp-Ht infected area. Statistics: means ± SEM; eight rats per group). ns, not significant; ** P
    Figure Legend Snippet: Exp-Htt-induced down-regulation of DARPP-32 is attenuated by MSK-1 overexpression 4 weeks post-infection. ( A ) The left striatum was infected with a lentiviral construct containing a gene fragment of huntingtin with 82 polyglutamine repeats (Exp-Htt) and the right striatum was infected with LV-Exp-Htt and another lentiviral construct containing the MSK-1 gene (Exp-Htt + MSK-1). ( B ) Immunocytochemical detection of Exp-Htt (upper panels), MSK-1 (middle panels) and DARPP-32 (lower panels) was performed with specific antibodies (see Materials and Methods), 4 weeks after infection. ( C ) The volume of striatal tissue showing Exp-Htt expression or DARPP-32 loss was measured on adjacent coronal sections across the rostro-caudal extension of the striatum. Seven to 10 sections per rat were analyzed (each section was 25 µm thick and was separated by 175 µm). ( D ) Graphical representation of the Exp-Htt volume expression in the rostro-caudal extension of the striatum. ( E ) Graphical representation of DARPP-32 loss/Exp-Ht infected area. Statistics: means ± SEM; eight rats per group). ns, not significant; ** P

    Techniques Used: Over Expression, Infection, Construct, Expressing

    MSK-1 controls histone H3 and CREB phosphorylation at the PGC-1α promoter: ( A ) Primary cultures of striatal neurons were transiently transfected with a cDNA encoding a GFP-tagged version of MSK-1 (yellow). After 16 h, PGC-1α expression was revealed immunocytochemically (red). Note that transfected neurons overexpressed PGC-1α (arrow). ( B ) Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of total RNA from striatal neurons isolated from WT and MSK-1 knock-out (MSK-KO) embryos, treated with glutamate (100 µ m ) for 1 or 2h. Primers amplifying PGC-1α and HPRT were used as internal controls. PGC-1α mRNA levels were quantified after normalization to the internal control levels (a.u.: arbitrary units). Chromatin immunoprecipitation (ChIP) followed by quantitative PCR using ( C ) primers encompassing the CRE binding site of the PGC-1α promoter, ( D ) primers located upstream this region. (C and D) Striatal neurons from WT embryos were treated (Glu) or not (Cont) with glutamate (100 µ m ) for 30 min. Immunoprecipitation was performed using specific antibodies against P-H3 (P-H3 Cont; P-H3 Glu), MSK-1 (MSK-1 Cont; MSK-1 Glu) and CREB (CREB Cont; CREB Glu). H4 immunoprecipitated chromatin was used as a negative control. The -fold increased binding on the PGC-1α promoter was determined after normalization to their appropriate input. Data are means ± SEM; three independent experiments, three points per sample in each experiment, *** P
    Figure Legend Snippet: MSK-1 controls histone H3 and CREB phosphorylation at the PGC-1α promoter: ( A ) Primary cultures of striatal neurons were transiently transfected with a cDNA encoding a GFP-tagged version of MSK-1 (yellow). After 16 h, PGC-1α expression was revealed immunocytochemically (red). Note that transfected neurons overexpressed PGC-1α (arrow). ( B ) Real-time quantitative polymerase chain reaction (RT-qPCR) analysis of total RNA from striatal neurons isolated from WT and MSK-1 knock-out (MSK-KO) embryos, treated with glutamate (100 µ m ) for 1 or 2h. Primers amplifying PGC-1α and HPRT were used as internal controls. PGC-1α mRNA levels were quantified after normalization to the internal control levels (a.u.: arbitrary units). Chromatin immunoprecipitation (ChIP) followed by quantitative PCR using ( C ) primers encompassing the CRE binding site of the PGC-1α promoter, ( D ) primers located upstream this region. (C and D) Striatal neurons from WT embryos were treated (Glu) or not (Cont) with glutamate (100 µ m ) for 30 min. Immunoprecipitation was performed using specific antibodies against P-H3 (P-H3 Cont; P-H3 Glu), MSK-1 (MSK-1 Cont; MSK-1 Glu) and CREB (CREB Cont; CREB Glu). H4 immunoprecipitated chromatin was used as a negative control. The -fold increased binding on the PGC-1α promoter was determined after normalization to their appropriate input. Data are means ± SEM; three independent experiments, three points per sample in each experiment, *** P

    Techniques Used: Pyrolysis Gas Chromatography, Transfection, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Isolation, Knock-Out, Chromatin Immunoprecipitation, Binding Assay, Immunoprecipitation, Negative Control

    MSK1 knock-out mice show spontaneous striatal degeneration. ( A ) Representative striatal sections from 9-month-old WT (left) and MSK-1 knock-out (MSK-1 KO, right) mice. Sections taken from the same rostro-caudal level were processed for cresyl violet staining after brain perfusion. Note the enlarged ventricles in MSK-1 KO mice. ( B ) Ventricle volume was measured from serial coronal sections (nine sections per animal) using ImageJ software. Data are means ± SEM; three WT and five MSK-KO mice; ** P
    Figure Legend Snippet: MSK1 knock-out mice show spontaneous striatal degeneration. ( A ) Representative striatal sections from 9-month-old WT (left) and MSK-1 knock-out (MSK-1 KO, right) mice. Sections taken from the same rostro-caudal level were processed for cresyl violet staining after brain perfusion. Note the enlarged ventricles in MSK-1 KO mice. ( B ) Ventricle volume was measured from serial coronal sections (nine sections per animal) using ImageJ software. Data are means ± SEM; three WT and five MSK-KO mice; ** P

    Techniques Used: Knock-Out, Mouse Assay, Staining, Software

    PGC-1α expression is induced by MSK-1 overexpression in the striatum. LV-Exp-Htt was injected in the left striatum (left panel) and both LV-Exp-Htt and LV-MSK-1 in the right striatum (right panel). Double staining for MSK-1 (red) and PGC-1α (green) was performed. At 4 weeks ( A ) and 10 weeks ( B ) post-infection, Exp-Htt down-regulated the expression of both MSK-1 and PGC-1α (left panels, asterisk). (A and B) In the co-infected sides, MSK-1 overexpression was associated with an increase in PGC-1α immunoreactivity at 4 weeks (A , right panels) and 10 weeks in the area surrounding the injection point ( B , right panels, triangle). ( C and D ) PGC-1α-immunoreactive neurons were counted in the infected area of the striatum at 4 (C) and 10 weeks (D), using the endogenous basal level of PGC-1α expression as threshold. Results are means ± SEM, eights rats per group; *** P
    Figure Legend Snippet: PGC-1α expression is induced by MSK-1 overexpression in the striatum. LV-Exp-Htt was injected in the left striatum (left panel) and both LV-Exp-Htt and LV-MSK-1 in the right striatum (right panel). Double staining for MSK-1 (red) and PGC-1α (green) was performed. At 4 weeks ( A ) and 10 weeks ( B ) post-infection, Exp-Htt down-regulated the expression of both MSK-1 and PGC-1α (left panels, asterisk). (A and B) In the co-infected sides, MSK-1 overexpression was associated with an increase in PGC-1α immunoreactivity at 4 weeks (A , right panels) and 10 weeks in the area surrounding the injection point ( B , right panels, triangle). ( C and D ) PGC-1α-immunoreactive neurons were counted in the infected area of the striatum at 4 (C) and 10 weeks (D), using the endogenous basal level of PGC-1α expression as threshold. Results are means ± SEM, eights rats per group; *** P

    Techniques Used: Pyrolysis Gas Chromatography, Expressing, Over Expression, Injection, Double Staining, Infection

    MSK-1 overexpression prevents Exp-Htt-induced neuronal dysfunction. ( A ) LV-Exp-Htt was injected in the left striatum and both LV-Exp-Htt and LV-MSK-1 were injected in the right striatum. Double staining for MSK-1 (red) and NeuN (green) was performed 10 weeks later. Left panels (Exp-Htt): note the disappearance of both MSK-1 and NeuN labeling within the dotted lines (injection point). Right panels (Exp-Htt + MSK-1): note the down-regulation of both markers within the dotted lines, despite the presence of some immunoreactive cells. Note also the overexpression of MSK-1 and NeuN in the area surrounding the injection point (outside the dotted lines). ( B ) Confocal micrograph showing the co-localization of aggregated Exp-Htt (green) and MSK-1 (red) in the area surrounding the injection point. ( C ) Cells overexpressing NeuN were quantified by image analysis, with a threshold based on mean endogenous NeuN expression. Cells exhibiting a fluorescence level above this threshold were considered positive, and were quantified in eight animals. ( D ) The mean nuclear area of cells in the area surrounding the injection point was analyzed with Hoechst staining and Image-ProPlus software. ** P
    Figure Legend Snippet: MSK-1 overexpression prevents Exp-Htt-induced neuronal dysfunction. ( A ) LV-Exp-Htt was injected in the left striatum and both LV-Exp-Htt and LV-MSK-1 were injected in the right striatum. Double staining for MSK-1 (red) and NeuN (green) was performed 10 weeks later. Left panels (Exp-Htt): note the disappearance of both MSK-1 and NeuN labeling within the dotted lines (injection point). Right panels (Exp-Htt + MSK-1): note the down-regulation of both markers within the dotted lines, despite the presence of some immunoreactive cells. Note also the overexpression of MSK-1 and NeuN in the area surrounding the injection point (outside the dotted lines). ( B ) Confocal micrograph showing the co-localization of aggregated Exp-Htt (green) and MSK-1 (red) in the area surrounding the injection point. ( C ) Cells overexpressing NeuN were quantified by image analysis, with a threshold based on mean endogenous NeuN expression. Cells exhibiting a fluorescence level above this threshold were considered positive, and were quantified in eight animals. ( D ) The mean nuclear area of cells in the area surrounding the injection point was analyzed with Hoechst staining and Image-ProPlus software. ** P

    Techniques Used: Over Expression, Injection, Double Staining, Labeling, Expressing, Fluorescence, Staining, Software

    32) Product Images from "GM1 ganglioside-independent intoxication by Cholera toxin"

    Article Title: GM1 ganglioside-independent intoxication by Cholera toxin

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006862

    CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p
    Figure Legend Snippet: CT induced ion secretion can be inhibited by pretreating the tissue with AAL and PNA. Human jejunal mucosae were pre-incubated with or without AAL or PNA at the indicated concentrations, mounted in an Ussing chamber and exposed to CT. (A) Dot plot showing percent difference in I ep to control tissue for jejunal mucosae over time. Each dot represents a mean of 4–7 donors (each treatment for each donor was tested in duplicates) with SEM error bars. Significance was calculated using a two-way-ANOVA with Tukey correction (compared to the CT). * represent CT to CT+AAL comparison and † represent CT to CT+PNA comparison (**** = p

    Techniques Used: Incubation

    33) Product Images from "Netrin-G/NGL Complexes Encode Functional Synaptic Diversification"

    Article Title: Netrin-G/NGL Complexes Encode Functional Synaptic Diversification

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1141-14.2014

    Altered PTP in netrin-G1-KO and netrin-G2-KO caused by distinct signaling deficits. A , B , PTP at the TA-CA1 synapses was impaired in netrin-G1-KO. In contrast, PTP at the SC-CA1 synapses was facilitated in netrin-G2-KO SC-CA1. C , D , fEPSP potentiation
    Figure Legend Snippet: Altered PTP in netrin-G1-KO and netrin-G2-KO caused by distinct signaling deficits. A , B , PTP at the TA-CA1 synapses was impaired in netrin-G1-KO. In contrast, PTP at the SC-CA1 synapses was facilitated in netrin-G2-KO SC-CA1. C , D , fEPSP potentiation

    Techniques Used:

    Deficiency of netrin-G2 attenuated synaptic transmission in the specific pathway. A , Schematics of field recording. Basal synaptic properties were examined in hippocampal CA1 and DG synapses. AMPAR-mediated fEPSPs were recorded by stimulating axon fibers:
    Figure Legend Snippet: Deficiency of netrin-G2 attenuated synaptic transmission in the specific pathway. A , Schematics of field recording. Basal synaptic properties were examined in hippocampal CA1 and DG synapses. AMPAR-mediated fEPSPs were recorded by stimulating axon fibers:

    Techniques Used: Transmission Assay

    Disappearance of presynaptic netrin-Gs in NGL-KO mice. A , B , Subregional iEM analyses for netrin-Gs in NGL-KO mice. Ultrathin sections from NGL1-KO and NGL2-KO mice were immunoreacted with antibodies against netrin-G1 and netrin-G2, respectively. C ,
    Figure Legend Snippet: Disappearance of presynaptic netrin-Gs in NGL-KO mice. A , B , Subregional iEM analyses for netrin-Gs in NGL-KO mice. Ultrathin sections from NGL1-KO and NGL2-KO mice were immunoreacted with antibodies against netrin-G1 and netrin-G2, respectively. C ,

    Techniques Used: Mouse Assay

    Presynaptic abnormalities in NGL-KO mice and normal dendritic morphologies of netrin-G2-KO and NGL2-KO mice. A–C , Synaptic phenotypes in TA-CA1 pathway of adult NGL1-KO mice. A , Normal basal synaptic transmission. B , Normal paired-pulse ratio
    Figure Legend Snippet: Presynaptic abnormalities in NGL-KO mice and normal dendritic morphologies of netrin-G2-KO and NGL2-KO mice. A–C , Synaptic phenotypes in TA-CA1 pathway of adult NGL1-KO mice. A , Normal basal synaptic transmission. B , Normal paired-pulse ratio

    Techniques Used: Mouse Assay, Transmission Assay

    34) Product Images from "Cell cycle-targeting microRNAs promote differentiation by enforcing cell-cycle exit"

    Article Title: Cell cycle-targeting microRNAs promote differentiation by enforcing cell-cycle exit

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1702914114

    CDK inhibition rescues the ciliogenesis defect in miR-34/449 TKO mice. ( A ) Histological sections of the nasal respiratory epithelium from wild-type and miR-34/449 TKO mice treated for 48 h with PBS, and miR-34/449 TKO mice treated for 48 h with ribociclib, stained for the proliferation marker Ki67 (brown). (Scale bars, 20 µm.) ( A , Right ) Quantification of the percentage of columnar epithelial cells with nuclear Ki67 staining (Ki67 + ) using sections from at least three mice per group. Data are presented as mean ± SD; *** P
    Figure Legend Snippet: CDK inhibition rescues the ciliogenesis defect in miR-34/449 TKO mice. ( A ) Histological sections of the nasal respiratory epithelium from wild-type and miR-34/449 TKO mice treated for 48 h with PBS, and miR-34/449 TKO mice treated for 48 h with ribociclib, stained for the proliferation marker Ki67 (brown). (Scale bars, 20 µm.) ( A , Right ) Quantification of the percentage of columnar epithelial cells with nuclear Ki67 staining (Ki67 + ) using sections from at least three mice per group. Data are presented as mean ± SD; *** P

    Techniques Used: Inhibition, Mouse Assay, Staining, Marker

    35) Product Images from "Sustained antibody responses depend on CD28 function in bone marrow-resident plasma cells"

    Article Title: Sustained antibody responses depend on CD28 function in bone marrow-resident plasma cells

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20110040

    Loss of CD28 ligands CD80 or CD86 diminishes long-lived antibody responses and long-lived PC numbers. (A) Mice were vaccinated and total serum IgG1 and NIP-IgG1 was analyzed by ELISA at the time points indicated. Each point represents one mouse, with the mean indicated by black bars. (B) Total PC numbers in spleen (left) and BM (right) was determined from total mononuclear cells by multiparametric flow cytometry using CD138 + B220 − to identify PC. Histograms are representative of three mice per group. Error bars represent the mean ± SD. (C and D) Splenic (C) and BM (D) NIP-IgG ASC numbers were determined by ELISPOT. Each point represents the triplicate mean of one mouse. Mean is indicated by black bars. ns, not significant. *, P
    Figure Legend Snippet: Loss of CD28 ligands CD80 or CD86 diminishes long-lived antibody responses and long-lived PC numbers. (A) Mice were vaccinated and total serum IgG1 and NIP-IgG1 was analyzed by ELISA at the time points indicated. Each point represents one mouse, with the mean indicated by black bars. (B) Total PC numbers in spleen (left) and BM (right) was determined from total mononuclear cells by multiparametric flow cytometry using CD138 + B220 − to identify PC. Histograms are representative of three mice per group. Error bars represent the mean ± SD. (C and D) Splenic (C) and BM (D) NIP-IgG ASC numbers were determined by ELISPOT. Each point represents the triplicate mean of one mouse. Mean is indicated by black bars. ns, not significant. *, P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunospot

    BMDCs interact with and support PC survival and function through CD28–CD80/CD86 interactions. (A) BM sections from WT mice were stained with antibodies against CD80 (red) and CD138 (green; image representative of four independent experiments). (B) Immunohistochemical staining from sternum sections of WT mice. Brown is fascin, identifying DCs, and pink is CD138 + PCs identified by arrows (image representative of two independent experiments). (C–H) Purified BM PCs were cocultured with BMDC of indicated genotypes for the indicated time periods. Total viable PC numbers were determined by 7AAD incorporation analyzed by flow cytometry (mean ± SD of three independent experiments is shown). Culture supernatants were analyzed for total IgG production by ELISA (mean ± SD is shown of one representative experiment of three). ns, not significant. *§, P
    Figure Legend Snippet: BMDCs interact with and support PC survival and function through CD28–CD80/CD86 interactions. (A) BM sections from WT mice were stained with antibodies against CD80 (red) and CD138 (green; image representative of four independent experiments). (B) Immunohistochemical staining from sternum sections of WT mice. Brown is fascin, identifying DCs, and pink is CD138 + PCs identified by arrows (image representative of two independent experiments). (C–H) Purified BM PCs were cocultured with BMDC of indicated genotypes for the indicated time periods. Total viable PC numbers were determined by 7AAD incorporation analyzed by flow cytometry (mean ± SD of three independent experiments is shown). Culture supernatants were analyzed for total IgG production by ELISA (mean ± SD is shown of one representative experiment of three). ns, not significant. *§, P

    Techniques Used: Mouse Assay, Staining, Immunohistochemistry, Purification, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    Loss of CD28 in the B lineage reduces long-lived antibody responses and long-lived PC numbers. (A) Experimental design. (B and C) Total splenic (B) or BM (C) mononuclear cells were analyzed in chimeras at time 0 before immunization for CD28 + CD3 + T cells (left) and CD28 + CD138 + B220 − PC (right) by multiparametric flow cytometry. (D) Total serum IgG1 and NIP-specific IgG1 was analyzed by ELISA at the time points indicated. Each point represents one mouse, with the mean indicated by black bars. (E) Total PC numbers in spleen (left) and BM (right) was determined from total mononuclear cells by multiparametric flow cytometry using CD138 + B220 − to identify PC. Histograms are representative of six mice (day 180 WT, n = 3) per group. Error bars represent the mean ± SD. (F and G) Splenic (F) and BM (G) NIP-IgG ASC numbers were determined by ELISPOT. Each point represents the triplicate mean of one mouse. Mean is indicated by black bars. ns, not significant. *, P
    Figure Legend Snippet: Loss of CD28 in the B lineage reduces long-lived antibody responses and long-lived PC numbers. (A) Experimental design. (B and C) Total splenic (B) or BM (C) mononuclear cells were analyzed in chimeras at time 0 before immunization for CD28 + CD3 + T cells (left) and CD28 + CD138 + B220 − PC (right) by multiparametric flow cytometry. (D) Total serum IgG1 and NIP-specific IgG1 was analyzed by ELISA at the time points indicated. Each point represents one mouse, with the mean indicated by black bars. (E) Total PC numbers in spleen (left) and BM (right) was determined from total mononuclear cells by multiparametric flow cytometry using CD138 + B220 − to identify PC. Histograms are representative of six mice (day 180 WT, n = 3) per group. Error bars represent the mean ± SD. (F and G) Splenic (F) and BM (G) NIP-IgG ASC numbers were determined by ELISPOT. Each point represents the triplicate mean of one mouse. Mean is indicated by black bars. ns, not significant. *, P

    Techniques Used: Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay, Mouse Assay, Enzyme-linked Immunospot

    36) Product Images from "Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey"

    Article Title: Origins of Cortical GABAergic Neurons in the Cynomolgus Monkey

    Journal: Cerebral Cortex (New York, NY)

    doi: 10.1093/cercor/bhn078

    Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).
    Figure Legend Snippet: Labeling patterns for Mash1 ( A ) and GAD65 ( B – E ) in coronal sections at E55. ( A ) IHC labeling for Mash1 is present in the GE but not in the neighboring structures including the caudate nucleus (caud), capsula interna (CI), and adjacent zones of the dorsal telencephalon: VZ, SVZ, and IZ. Arrows indicated border between GE and adjacent structures. ( B ) A high level of GAD65 labeling is present in the GE. In addition a stream of postmitotic like GAD65-containing neurons extending from GE to dorsal telencephalon was observed in the lower IZ/upper SVZ. Arrows indicate the deeper border of this stream, showing that at E55 none of the cells leaving GE migrate through VZ and lower SVZ. Boxes: magnified regions in ( C ) and ( D ). ( C ) GAD65-containing cells with a characteristic morphology of postmitotic migrating neurons, including a leading process (arrows) leaving the GE. ( D ) In the IZ and upper SVZ of dorsal telencephalon, the majority of GAD65 migrating neurons are tangentially (parallel to pia) oriented (arrows). ( E ) Tangentially oriented GAD65 migrating neurons are also observed in the SP (arrows), CP, and MZ of the dorsal telencephalon. Scale bars = 350 μm ( A , B ), 20 μm, ( C , D , E ).

    Techniques Used: Labeling, Immunohistochemistry

    37) Product Images from "Colocalization of GPR120 and anterior pituitary hormone-producing cells in female Japanese Black cattle"

    Article Title: Colocalization of GPR120 and anterior pituitary hormone-producing cells in female Japanese Black cattle

    Journal: The Journal of Reproduction and Development

    doi: 10.1262/jrd.2019-111

    Dual immunohistochemistry of GPR120 (red) and luteinizing hormone β subunit (LHβ) (green), follicle-stimulating hormone β subunit (FSHβ) (green), thyroid-stimulating hormone β subunit (TSHβ) (green), prolactin (PRL) (green), growth hormone (GH) (green), and adrenocorticotropic hormone (ACTH) (green) with 4',6-diaminodino-2-phenylinodole (DAPI) (blue) in anterior pituitary gland of ovariectomized heifers. White and black arrowheads indicate representative dual-labeled cells and single-labeled cells, respectively. Scale bar = 30 µm.
    Figure Legend Snippet: Dual immunohistochemistry of GPR120 (red) and luteinizing hormone β subunit (LHβ) (green), follicle-stimulating hormone β subunit (FSHβ) (green), thyroid-stimulating hormone β subunit (TSHβ) (green), prolactin (PRL) (green), growth hormone (GH) (green), and adrenocorticotropic hormone (ACTH) (green) with 4',6-diaminodino-2-phenylinodole (DAPI) (blue) in anterior pituitary gland of ovariectomized heifers. White and black arrowheads indicate representative dual-labeled cells and single-labeled cells, respectively. Scale bar = 30 µm.

    Techniques Used: Immunohistochemistry, Labeling

    38) Product Images from "Low-glucose enhances keratocyte-characteristic phenotype from corneal stromal cells in serum-free conditions"

    Article Title: Low-glucose enhances keratocyte-characteristic phenotype from corneal stromal cells in serum-free conditions

    Journal: Scientific Reports

    doi: 10.1038/srep10839

    Effects of glucose concentration on the expression of keratocyte-characteristic protein markers. a ) The expression levels of CD34, keratocan, and αSMA protein markers was analysed from lysates obtained from human corneal tissue (hC) and human corneal stromal cells grown in LG, HG, or serum-containing BM by immunoblotting. Detection of α-tubulin was performed to exclude variations from total protein load. Migration was evaluated using pre-stained molecular ladder (MW) to distinguish between high- ( white ) and low-migratory protein forms ( black arrowhead ). Panels show representative images from three independent experiments. b ) The expression of CD34 (i), keratocan (ii), and αSMA (iii) was quantified by densitometry, normalized with α-tubulin signal, and compared between LG ( light blue ) and HG ( dark blue bars ), with expression from human corneal tissue (hC, white ) and cells grown in BM ( grey bars ) used as reference. Data was expressed as average ± S.D. of three independent experiments ( n = 3); * corresponded to p
    Figure Legend Snippet: Effects of glucose concentration on the expression of keratocyte-characteristic protein markers. a ) The expression levels of CD34, keratocan, and αSMA protein markers was analysed from lysates obtained from human corneal tissue (hC) and human corneal stromal cells grown in LG, HG, or serum-containing BM by immunoblotting. Detection of α-tubulin was performed to exclude variations from total protein load. Migration was evaluated using pre-stained molecular ladder (MW) to distinguish between high- ( white ) and low-migratory protein forms ( black arrowhead ). Panels show representative images from three independent experiments. b ) The expression of CD34 (i), keratocan (ii), and αSMA (iii) was quantified by densitometry, normalized with α-tubulin signal, and compared between LG ( light blue ) and HG ( dark blue bars ), with expression from human corneal tissue (hC, white ) and cells grown in BM ( grey bars ) used as reference. Data was expressed as average ± S.D. of three independent experiments ( n = 3); * corresponded to p

    Techniques Used: Concentration Assay, Expressing, Migration, Staining

    39) Product Images from "Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone"

    Article Title: Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone

    Journal: Scientific Reports

    doi: 10.1038/srep35599

    HE staining and B220 and IgG deposit detection on testicular sections from control ( a–d ) and TGC-induced EAO ( e–h ) mice. Testes histological sections from control ( a , b ) and TGC-induced EAO ( e , f ) mice were stained with hematoxylin and eosin. Additional tissue sections were incubated with specific antibodies to detect B220 ( c , g ) and IgG deposits ( d , h ) in testes from control ( c , d ) and EAO ( g , h ) mice. The presence of infiltrating B-cells ( g ) and IgG deposits ( h ) with disrupted spermatogenesis was observed in the testes from TGC-induced EAO mice. Brown spots indicate positive cells ( g , h ). Scale bar: 150 μm ( a – f ).
    Figure Legend Snippet: HE staining and B220 and IgG deposit detection on testicular sections from control ( a–d ) and TGC-induced EAO ( e–h ) mice. Testes histological sections from control ( a , b ) and TGC-induced EAO ( e , f ) mice were stained with hematoxylin and eosin. Additional tissue sections were incubated with specific antibodies to detect B220 ( c , g ) and IgG deposits ( d , h ) in testes from control ( c , d ) and EAO ( g , h ) mice. The presence of infiltrating B-cells ( g ) and IgG deposits ( h ) with disrupted spermatogenesis was observed in the testes from TGC-induced EAO mice. Brown spots indicate positive cells ( g , h ). Scale bar: 150 μm ( a – f ).

    Techniques Used: Staining, Mouse Assay, Incubation

    40) Product Images from "B Cell Response and Hemagglutinin Stalk-Reactive Antibody Production in Different Age Cohorts following 2009 H1N1 Influenza Virus Vaccination"

    Article Title: B Cell Response and Hemagglutinin Stalk-Reactive Antibody Production in Different Age Cohorts following 2009 H1N1 Influenza Virus Vaccination

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00735-12

    Circulating ASCs induced by pH1N1 vaccination. B cells enriched from blood were analyzed by an ELISpot assay for vaccine-specific ASCs and by flow cytometry for ASC phenotypes. (A to C) Vaccine-specific ASCs. IgG and IgA ASC frequencies on days 6 to 8
    Figure Legend Snippet: Circulating ASCs induced by pH1N1 vaccination. B cells enriched from blood were analyzed by an ELISpot assay for vaccine-specific ASCs and by flow cytometry for ASC phenotypes. (A to C) Vaccine-specific ASCs. IgG and IgA ASC frequencies on days 6 to 8

    Techniques Used: Enzyme-linked Immunospot, Flow Cytometry, Cytometry

    Relationship between vaccine-specific IgG ASCs and the HAI response. A linear regression model assuming different coefficients for different age cohorts was fitted to evaluate the relationship between the frequency (log 10) of circulating vaccine-specific
    Figure Legend Snippet: Relationship between vaccine-specific IgG ASCs and the HAI response. A linear regression model assuming different coefficients for different age cohorts was fitted to evaluate the relationship between the frequency (log 10) of circulating vaccine-specific

    Techniques Used:

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    PTPN11 D61G overexpression increases surface AMPA receptor expression a. and b. Representative images of surface GluA1 staining in cultured neurons. GFP alone (a) or PTPN11 D61G and GFP (b) were co–expressed using a bicistronic Sindbis viral vector in cultured hippocampal neurons (DIV21). Scale, 20μm. c. Representative images of western blotting of total and <t>biotinylated</t> surface proteins. Cadherin and Rab-4 were used as markers for surface and cytosol expression, respectively. Full-length blots/gels are presented in Supplementary Figure 11 . d. Surface expression of GluA1 was significantly increased in PTPN11 D61G expressing neurons compared to WT PTPN11 expressing neurons, while the total expression level of GluA1 did not differ between WT PTPN11 and PTPN11 D61G transfected neurons.
    Biotinylated Anti Rabbit Antibody, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PTPN11 D61G overexpression increases surface AMPA receptor expression a. and b. Representative images of surface GluA1 staining in cultured neurons. GFP alone (a) or PTPN11 D61G and GFP (b) were co–expressed using a bicistronic Sindbis viral vector in cultured hippocampal neurons (DIV21). Scale, 20μm. c. Representative images of western blotting of total and <t>biotinylated</t> surface proteins. Cadherin and Rab-4 were used as markers for surface and cytosol expression, respectively. Full-length blots/gels are presented in Supplementary Figure 11 . d. Surface expression of GluA1 was significantly increased in PTPN11 D61G expressing neurons compared to WT PTPN11 expressing neurons, while the total expression level of GluA1 did not differ between WT PTPN11 and PTPN11 D61G transfected neurons.
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    PTPN11 D61G overexpression increases surface AMPA receptor expression a. and b. Representative images of surface GluA1 staining in cultured neurons. GFP alone (a) or PTPN11 D61G and GFP (b) were co–expressed using a bicistronic Sindbis viral vector in cultured hippocampal neurons (DIV21). Scale, 20μm. c. Representative images of western blotting of total and biotinylated surface proteins. Cadherin and Rab-4 were used as markers for surface and cytosol expression, respectively. Full-length blots/gels are presented in Supplementary Figure 11 . d. Surface expression of GluA1 was significantly increased in PTPN11 D61G expressing neurons compared to WT PTPN11 expressing neurons, while the total expression level of GluA1 did not differ between WT PTPN11 and PTPN11 D61G transfected neurons.

    Journal: Nature neuroscience

    Article Title: Mechanism and treatment for the learning and memory deficits associated with mouse models of Noonan syndrome

    doi: 10.1038/nn.3863

    Figure Lengend Snippet: PTPN11 D61G overexpression increases surface AMPA receptor expression a. and b. Representative images of surface GluA1 staining in cultured neurons. GFP alone (a) or PTPN11 D61G and GFP (b) were co–expressed using a bicistronic Sindbis viral vector in cultured hippocampal neurons (DIV21). Scale, 20μm. c. Representative images of western blotting of total and biotinylated surface proteins. Cadherin and Rab-4 were used as markers for surface and cytosol expression, respectively. Full-length blots/gels are presented in Supplementary Figure 11 . d. Surface expression of GluA1 was significantly increased in PTPN11 D61G expressing neurons compared to WT PTPN11 expressing neurons, while the total expression level of GluA1 did not differ between WT PTPN11 and PTPN11 D61G transfected neurons.

    Article Snippet: A biotinylated anti-rabbit antibody (1:50, 1 h at room temperature; Vector laboratories) was used as a secondary, which was followed by avidin-biotin-peroxidase complex (Vector Laboratories) formation for 30 min.

    Techniques: Over Expression, Expressing, Staining, Cell Culture, Plasmid Preparation, Western Blot, Transfection