nickel enhanced dab  (Vector Laboratories)


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    Biotinylated Horse Anti Mouse IgG Antibody rat adsorbed
    Description:
    Biotinylated Horse Anti Mouse IgG Antibody rat adsorbed 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 Horse Anti Mouse IgG rat adsorbed is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 100 H L indicates the antibody recognizes both heavy and light chains This specially prepared antibody is designed to be used on tissues that may contain interfering endogenous immunoglobulins The rat adsorbed anti mouse IgG H L products are designed for use with rat tissues that may contain endogenous rat immunoglobulins This adsorbed antibody allows primary antibodies made in mouse to be used on rat tissues
    Catalog Number:
    ba-2001
    Price:
    None
    Host:
    Horse
    Size:
    0 5 mg
    Category:
    Antibodies
    Reactivity:
    Mouse
    Buy from Supplier


    Structured Review

    Vector Laboratories nickel enhanced dab
    Biotinylated Horse Anti Mouse IgG Antibody rat adsorbed
    Biotinylated Horse Anti Mouse IgG Antibody rat adsorbed 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 Horse Anti Mouse IgG rat adsorbed is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 100 H L indicates the antibody recognizes both heavy and light chains This specially prepared antibody is designed to be used on tissues that may contain interfering endogenous immunoglobulins The rat adsorbed anti mouse IgG H L products are designed for use with rat tissues that may contain endogenous rat immunoglobulins This adsorbed antibody allows primary antibodies made in mouse to be used on rat tissues
    https://www.bioz.com/result/nickel enhanced dab/product/Vector Laboratories
    Average 88 stars, based on 3305 article reviews
    Price from $9.99 to $1999.99
    nickel enhanced dab - by Bioz Stars, 2021-02
    88/100 stars

    Images

    1) Product Images from "Parecoxib is neuroprotective in spontaneously hypertensive rats after transient middle cerebral artery occlusion: a divided treatment response?"

    Article Title: Parecoxib is neuroprotective in spontaneously hypertensive rats after transient middle cerebral artery occlusion: a divided treatment response?

    Journal: Journal of Neuroinflammation

    doi: 10.1186/1742-2094-3-31

    COX-2 and NeuN double stains 24 hours and one week after tMCAo . The COX-2 IHC was developed with nickel-enhanced DAB (black), whereas NeuN was visualized with NovaRed ® (brownish red). The images 5A and 5B are obtained from a pilot study where the animal was euthanized 24 hours after tMCAo. 5A visualizes a relatively small neocortical infarct in the right hemisphere. The box delineates a part of the ischemic border zone that is shown at forty times magnification in 5B . The penumbra contains large swollen neurons that express the membrane-bound COX-2 enzyme. In the infarct core the neurons tend to be small and star-shaped due to irreversible neuronal death. 5C and 5E are from a saline-treated animal one week after tMCAo. Forty times magnifications of the boxes are shown in 5D and 5F . The neurons in the border zone on Day 8 after ischemic injury showed a perinuclear expression pattern of the COX-2 enzyme ( 5D ). COX-2 + neurons can be found in areas like the neocortex, piriform cortex and the DG of the hippocampus under normal conditions. 5F shows COX-2 expressed in dendrites of neurons in the molecular cell layer of the DG. The scale bar in 5A is 5 mm, whereas the scale bars in 5B , 5D and 5F equals 50 μm.
    Figure Legend Snippet: COX-2 and NeuN double stains 24 hours and one week after tMCAo . The COX-2 IHC was developed with nickel-enhanced DAB (black), whereas NeuN was visualized with NovaRed ® (brownish red). The images 5A and 5B are obtained from a pilot study where the animal was euthanized 24 hours after tMCAo. 5A visualizes a relatively small neocortical infarct in the right hemisphere. The box delineates a part of the ischemic border zone that is shown at forty times magnification in 5B . The penumbra contains large swollen neurons that express the membrane-bound COX-2 enzyme. In the infarct core the neurons tend to be small and star-shaped due to irreversible neuronal death. 5C and 5E are from a saline-treated animal one week after tMCAo. Forty times magnifications of the boxes are shown in 5D and 5F . The neurons in the border zone on Day 8 after ischemic injury showed a perinuclear expression pattern of the COX-2 enzyme ( 5D ). COX-2 + neurons can be found in areas like the neocortex, piriform cortex and the DG of the hippocampus under normal conditions. 5F shows COX-2 expressed in dendrites of neurons in the molecular cell layer of the DG. The scale bar in 5A is 5 mm, whereas the scale bars in 5B , 5D and 5F equals 50 μm.

    Techniques Used: Immunohistochemistry, Expressing

    BrdU, ED-1 and NeuN stains one week after tMCAo . 9A to 9F show the IHC for BrdU and ED-1. BrdU was developed with nickel-enhanced DAB (black), whereas the activated microglia marker ED-1 was visualized with NovaRed ® (brownish red). 9A shows a representative example of a tMCAo animal one week after surgery. Four and forty time magnifications of the boxes in 9A and 9B are shown in 9B and 9C , respectively. The BrdU + cells are typically found in clusters in the subgranular cell layer in DG. Our counting procedure started with delineating the DG. Hereafter, the BrdU + cells in the whole DG were counted at forty times magnification ( 9C ). We generally observed a very scarce ED-1 expression in the DG unless the hippocampus was directly affected by ischemic injury. 9D to 9I show an example of ischemia affecting the right hippocampus. The boxed area in 9D is shown at higher magnification in 9E . Activated microglia was abundantly seen in a part of the CA-1 and the whole DG. 9F shows a forty time magnification of the box in 9E . Clearly, activated microglia had an intimate relation to an increasing number of BrdU + cells. Panel 9G to 9I show NeuN IHC developed with nickel-enhanced DAB. 9G and 9H correspond to 9D and 9E . The boxed area in 9H of the CA-1 is shown at forty time magnification in 9I . Note the ischemic degeneration of this part of the CA-1. The scale bar in 9A is 5 mm. In 9B the scale bar represents 300 mm, and in 9E and 9H 1 mm. The scale bars in 9C , 9F and 9I equals 50 μm.
    Figure Legend Snippet: BrdU, ED-1 and NeuN stains one week after tMCAo . 9A to 9F show the IHC for BrdU and ED-1. BrdU was developed with nickel-enhanced DAB (black), whereas the activated microglia marker ED-1 was visualized with NovaRed ® (brownish red). 9A shows a representative example of a tMCAo animal one week after surgery. Four and forty time magnifications of the boxes in 9A and 9B are shown in 9B and 9C , respectively. The BrdU + cells are typically found in clusters in the subgranular cell layer in DG. Our counting procedure started with delineating the DG. Hereafter, the BrdU + cells in the whole DG were counted at forty times magnification ( 9C ). We generally observed a very scarce ED-1 expression in the DG unless the hippocampus was directly affected by ischemic injury. 9D to 9I show an example of ischemia affecting the right hippocampus. The boxed area in 9D is shown at higher magnification in 9E . Activated microglia was abundantly seen in a part of the CA-1 and the whole DG. 9F shows a forty time magnification of the box in 9E . Clearly, activated microglia had an intimate relation to an increasing number of BrdU + cells. Panel 9G to 9I show NeuN IHC developed with nickel-enhanced DAB. 9G and 9H correspond to 9D and 9E . The boxed area in 9H of the CA-1 is shown at forty time magnification in 9I . Note the ischemic degeneration of this part of the CA-1. The scale bar in 9A is 5 mm. In 9B the scale bar represents 300 mm, and in 9E and 9H 1 mm. The scale bars in 9C , 9F and 9I equals 50 μm.

    Techniques Used: Immunohistochemistry, Marker, Expressing

    2) Product Images from "Aged chimpanzees exhibit pathologic hallmarks of Alzheimer’s disease"

    Article Title: Aged chimpanzees exhibit pathologic hallmarks of Alzheimer’s disease

    Journal: Neurobiology of aging

    doi: 10.1016/j.neurobiolaging.2017.07.006

    APP/Aβ and Aβ42-ir plaques and vessels in aged chimpanzees (subject 12: A-B,D-F,H,M-O; subject 9: C; subject 20: G,J,L; subject 19: I,K): (A-B,E) APP/Aβ-ir plaques, (C-D,F) Aβ42-ir plaques, (G) APP/Aβ-ir cortical artery and arterioles, (H) APP/Aβ-ir leptomeningeal arteries, (I) Aβ42-ir cortical artery (J) Aβ42-ir cortical arterioles, (K) APP/Aβ-ir cortical arterioles, (L) Aβ42-ir cortical arterioles, and (M-O) APP/Aβ-ir plaques (black arrows) near immunopositive vessels (white arrows). Scale bars = 250 µm (A, D, G, J-L, M-O) or 25 µm (B-C, E-F, H-I).
    Figure Legend Snippet: APP/Aβ and Aβ42-ir plaques and vessels in aged chimpanzees (subject 12: A-B,D-F,H,M-O; subject 9: C; subject 20: G,J,L; subject 19: I,K): (A-B,E) APP/Aβ-ir plaques, (C-D,F) Aβ42-ir plaques, (G) APP/Aβ-ir cortical artery and arterioles, (H) APP/Aβ-ir leptomeningeal arteries, (I) Aβ42-ir cortical artery (J) Aβ42-ir cortical arterioles, (K) APP/Aβ-ir cortical arterioles, (L) Aβ42-ir cortical arterioles, and (M-O) APP/Aβ-ir plaques (black arrows) near immunopositive vessels (white arrows). Scale bars = 250 µm (A, D, G, J-L, M-O) or 25 µm (B-C, E-F, H-I).

    Techniques Used:

    Plaque and vessel volumes in 20 chimpanzees aged 37–62 years old (mean age = 46 years): (A) Average total plaque and vessel volume (%) (r s = 0.70, p ≤ 0.01, small circles represent outliers), (B) correlation of total APP/Aβ plaque volume vs total Aβ42 plaque volume across brain regions (R 2 = 0.54, p
    Figure Legend Snippet: Plaque and vessel volumes in 20 chimpanzees aged 37–62 years old (mean age = 46 years): (A) Average total plaque and vessel volume (%) (r s = 0.70, p ≤ 0.01, small circles represent outliers), (B) correlation of total APP/Aβ plaque volume vs total Aβ42 plaque volume across brain regions (R 2 = 0.54, p

    Techniques Used:

    Significant correlations of APP/Aβ (%) and tau (mm 3 ) lesions (p’s ≤ 0.05): (A) HC pretangle density vs NC and HC APP/Aβ vessel volume (NC: R 2 = 0.22, HC: R 2 = 0.18), (B) HC pretangle density vs HC APP/Aβ plaque volume (R 2 = 0.19), (C) HC NFT density vs NC APP/Aβ vessel volume (R 2 = 0.23), (D) HC AT8-ir tau NC density vs NC and HC APP/Aβ vessel volume (NC: R 2 = 0.20, HC: R 2 = 0.20), and (E) NC AT8-ir tau NC density and HC APP/Aβ plaque volume (R 2 = 0.27).
    Figure Legend Snippet: Significant correlations of APP/Aβ (%) and tau (mm 3 ) lesions (p’s ≤ 0.05): (A) HC pretangle density vs NC and HC APP/Aβ vessel volume (NC: R 2 = 0.22, HC: R 2 = 0.18), (B) HC pretangle density vs HC APP/Aβ plaque volume (R 2 = 0.19), (C) HC NFT density vs NC APP/Aβ vessel volume (R 2 = 0.23), (D) HC AT8-ir tau NC density vs NC and HC APP/Aβ vessel volume (NC: R 2 = 0.20, HC: R 2 = 0.20), and (E) NC AT8-ir tau NC density and HC APP/Aβ plaque volume (R 2 = 0.27).

    Techniques Used:

    3) 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

    4) Product Images from "A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71"

    Article Title: A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71

    Journal: Journal of Virology

    doi: 10.1128/JVI.00183-19

    Replication of EV71 in orally infected Tg mice. (A) Histopathological changes and expression of EV71 VP2 protein in infected mice. Two-week-old Tg mice were inoculated i.g. with the EV71-4643 strain and euthanized at 6 days postinfection. (Left) Cerebral cortex, hippocampus, cerebellum, thoracic spinal cord, pons, medulla, skeletal muscle (mice with hind-limb paralysis and moribund mice with complete paralysis), and hind-limb paw of Tg mice were retrieved for sample processing, IHC staining with the anti-EV71 VP2 antibody, and hematoxylin counterstaining. (Right) In parallel, sections were stained with H E. Original magnification, ×200. A representative result out of six experiments is shown. (B) Viral titers in various parts of the CNS and GI tract and in skeletal muscle of infected mice. Two-week-old Tg and non-Tg mice were infected i.g. with 1 × 10 8 PFU of the EV71-4643 strain and euthanized at the indicated times. Brains and GI tracts were collected and dissected into parts, as indicated in the figure. The anterior small intestine includes the duodenum and parts of the jejunum, while the posterior small intestine includes parts of the jejunum and ileum. Viral titers were determined using a plaque-forming assay and are expressed as log PFU per gram of tissue. ns, not significant; *, P
    Figure Legend Snippet: Replication of EV71 in orally infected Tg mice. (A) Histopathological changes and expression of EV71 VP2 protein in infected mice. Two-week-old Tg mice were inoculated i.g. with the EV71-4643 strain and euthanized at 6 days postinfection. (Left) Cerebral cortex, hippocampus, cerebellum, thoracic spinal cord, pons, medulla, skeletal muscle (mice with hind-limb paralysis and moribund mice with complete paralysis), and hind-limb paw of Tg mice were retrieved for sample processing, IHC staining with the anti-EV71 VP2 antibody, and hematoxylin counterstaining. (Right) In parallel, sections were stained with H E. Original magnification, ×200. A representative result out of six experiments is shown. (B) Viral titers in various parts of the CNS and GI tract and in skeletal muscle of infected mice. Two-week-old Tg and non-Tg mice were infected i.g. with 1 × 10 8 PFU of the EV71-4643 strain and euthanized at the indicated times. Brains and GI tracts were collected and dissected into parts, as indicated in the figure. The anterior small intestine includes the duodenum and parts of the jejunum, while the posterior small intestine includes parts of the jejunum and ileum. Viral titers were determined using a plaque-forming assay and are expressed as log PFU per gram of tissue. ns, not significant; *, P

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

    5) Product Images from "Breast Cancer: An Examination of the Potential of ACKR3 to Modify the Response of CXCR4 to CXCL12"

    Article Title: Breast Cancer: An Examination of the Potential of ACKR3 to Modify the Response of CXCR4 to CXCL12

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19113592

    CXCR4 and ACKR3 staining using IHC in breast cancer tissue. 4 μm sections from human breast cancer were stained for CXCR4 (1:40) and ACKR3 (1:100) using immunohistochemistry following no pre-treatment or EDTA antigen retrieval pre-treatment, respectively. Briefly, protocol from the VECTASTAIN ABC HRP kit was followed, signal was developed using DAB and counterstained with haematoxylin. No primary antibody was used as a control. n = 2.
    Figure Legend Snippet: CXCR4 and ACKR3 staining using IHC in breast cancer tissue. 4 μm sections from human breast cancer were stained for CXCR4 (1:40) and ACKR3 (1:100) using immunohistochemistry following no pre-treatment or EDTA antigen retrieval pre-treatment, respectively. Briefly, protocol from the VECTASTAIN ABC HRP kit was followed, signal was developed using DAB and counterstained with haematoxylin. No primary antibody was used as a control. n = 2.

    Techniques Used: Staining, Immunohistochemistry

    6) Product Images from "CXCL9 and 11 in patients with pulmonary sarcoidosis: a role of alveolar macrophages"

    Article Title: CXCL9 and 11 in patients with pulmonary sarcoidosis: a role of alveolar macrophages

    Journal: Clinical and Experimental Immunology

    doi: 10.1111/j.1365-2249.2007.03423.x

    Immunostaining of monokine induced by interferon γ (Mig)/CXCL9, interferon-inducible protein-10 (IP-10)/CXCL10 and interferon-inducible T cell α-chemoattractant (I-TAC)/CXCL11 in bronchoalveolar lavage (BA) cells and sarcoid lungs. BAL cells were cytocentrifuged on glass slides and fixed. The slides were incubated with control rabbit serum (a), anti-Mig/CXCL9 (b), IP-10/CXCL10 (c) or I-TAC/CXCL11 antibody (d), and stained with Alexa fluor 488-conjugated goat anti-rabbit IgG (H + l) antibody (green fluorescence) and 4′,6 diamidino-2-phenylindole (DAPI; by which nuclei were counterstained) (blue). Immunostaining was visualized using a fluorescence microscope (original magnifications: × 400). Arrows point to the lymphocytes which are negative for CXCR3 ligands. Freshly frozen lung sections derived from three patients with stage II of sarcoidosis were also stained with control rabbit serum (e), anti-Mig/CXCL9 (f), IP-10/CXCL10 (g) or I-TAC/CXCL11 antibody (h) and RTU Vectastain Universal Quick Kit, and developed with diaminobenzidine (DAB) substrate (original magnifications: × 200). The insets in e, f, g and h show the staining of macrophages in the alveolar lumen (original magnifications: × 400). Scale bars = 50 µm.
    Figure Legend Snippet: Immunostaining of monokine induced by interferon γ (Mig)/CXCL9, interferon-inducible protein-10 (IP-10)/CXCL10 and interferon-inducible T cell α-chemoattractant (I-TAC)/CXCL11 in bronchoalveolar lavage (BA) cells and sarcoid lungs. BAL cells were cytocentrifuged on glass slides and fixed. The slides were incubated with control rabbit serum (a), anti-Mig/CXCL9 (b), IP-10/CXCL10 (c) or I-TAC/CXCL11 antibody (d), and stained with Alexa fluor 488-conjugated goat anti-rabbit IgG (H + l) antibody (green fluorescence) and 4′,6 diamidino-2-phenylindole (DAPI; by which nuclei were counterstained) (blue). Immunostaining was visualized using a fluorescence microscope (original magnifications: × 400). Arrows point to the lymphocytes which are negative for CXCR3 ligands. Freshly frozen lung sections derived from three patients with stage II of sarcoidosis were also stained with control rabbit serum (e), anti-Mig/CXCL9 (f), IP-10/CXCL10 (g) or I-TAC/CXCL11 antibody (h) and RTU Vectastain Universal Quick Kit, and developed with diaminobenzidine (DAB) substrate (original magnifications: × 200). The insets in e, f, g and h show the staining of macrophages in the alveolar lumen (original magnifications: × 400). Scale bars = 50 µm.

    Techniques Used: Immunostaining, Incubation, Staining, Fluorescence, Microscopy, Derivative Assay

    7) Product Images from "Cell-based quantification of molecular biomarkers in histopathology specimens"

    Article Title: Cell-based quantification of molecular biomarkers in histopathology specimens

    Journal: Histopathology

    doi: 10.1111/j.1365-2559.2011.03878.x

    Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in human breast carcinoma cells. Sections of two different breast tumours were stained and analysed as described for Figure 5. Brightfield images of the two different tumours are shown ( A,G ), with the unmixed channels for 3,3-diaminobenzidine (Ki67) ( B,H ) and SG Blue (p-ERK) ( C,I ). Composite images showing whole-cell segmentation of the tumour (cytokeratin-positive) cells are shown ( D,J ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for tumour cells ( E,K ) and for non-tumour (stromal) cells ( F,L ), with each dot representing one cell.
    Figure Legend Snippet: Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in human breast carcinoma cells. Sections of two different breast tumours were stained and analysed as described for Figure 5. Brightfield images of the two different tumours are shown ( A,G ), with the unmixed channels for 3,3-diaminobenzidine (Ki67) ( B,H ) and SG Blue (p-ERK) ( C,I ). Composite images showing whole-cell segmentation of the tumour (cytokeratin-positive) cells are shown ( D,J ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for tumour cells ( E,K ) and for non-tumour (stromal) cells ( F,L ), with each dot representing one cell.

    Techniques Used: Immunostaining, Staining

    Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in lymphoid cells in a human breast carcinoma. A section of a breast tumour was stained sequentially with anti-p-ERK (SG Blue), anti-Ki67 [3,3-diaminobenzidine (DAB)] and anti-CK (Alexa-488) antibodies, and this was followed by haematoxylin staining, multispectral imaging (×400), and cytometric analysis. The brightfield image of a lymphoid nodule in the tumour is shown ( A ), along with the unmixed channels for DAB (Ki67) ( B ), SG Blue (p-ERK) ( C ), and Alexa-488 (cytokeratin) ( D ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for cells in the lymphoid nodule ( E ) and for tumour cells ( F ), with each dot representing one cell.
    Figure Legend Snippet: Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in lymphoid cells in a human breast carcinoma. A section of a breast tumour was stained sequentially with anti-p-ERK (SG Blue), anti-Ki67 [3,3-diaminobenzidine (DAB)] and anti-CK (Alexa-488) antibodies, and this was followed by haematoxylin staining, multispectral imaging (×400), and cytometric analysis. The brightfield image of a lymphoid nodule in the tumour is shown ( A ), along with the unmixed channels for DAB (Ki67) ( B ), SG Blue (p-ERK) ( C ), and Alexa-488 (cytokeratin) ( D ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for cells in the lymphoid nodule ( E ) and for tumour cells ( F ), with each dot representing one cell.

    Techniques Used: Immunostaining, Staining, Imaging

    8) Product Images from "Sox-2 Positive Neural Progenitors in the Primate Striatum Undergo Dynamic Changes after Dopamine Denervation"

    Article Title: Sox-2 Positive Neural Progenitors in the Primate Striatum Undergo Dynamic Changes after Dopamine Denervation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066377

    Ultra-structural analysis of Sox-2 expression. A. Sox-2 immuno-gold label is observed in the nucleus (arrowheads) in ependymal cells and astrocytes in ultra-thin sections of the SVZ. Boxed areas show an ependymal cell (E) and an astrocyte (As). Ependymal layer (EL), gap layer (HL), astrocyte ribbon layer (R). Scale bar = 20 um; Scale bar in boxed areas = 1 um. a1. Magnification shows a Sox-2 + astrocyte and lack of immunoreactivity in an oligodendrocyte (O) and a microglial cell (Mi). Astrocytes are recognized by the presence of dense bundles of intermediate filaments (arrows). Scale bar = 2 um. B. Sox-2 immuno-gold staining in a group of neuroblasts (type A cells). Scale bar = 20 um. b1. Magnification of three neuroblasts showing the nuclear staining. Neuroblasts form chain-like structures with characteristic intercellular spaces (arrows). Scale bar = 2 um. C. Light microscopy image of a semi-thin section showing the distribution and characterization of cells expressing Sox-2 in the dentate gyrus. Sox-2 + cells are visualized by the nuclear dark brown DAB precipitate. The section was counterstained with toluidine blue. The line marks the boundary between the granular layer (GrL) and the hilus (H) where most labeled cells were located. Scale bar = 100 um. c1 ) Neurons in the granular layer were not Sox-2 + . Scale bar = 10 um; Insert = 1 um. D. Microphotograph of a striatal field of a control monkey showing a Sox-2 + astrocyte and unlabeled neuron (Neu) and microglial cells (Mi). Scale bar = 5 um. d1. Boxed area of the astrocyte (As) nucleus shows the gold particles (arrowheads). Scale bar = 1 um. E. A representative Sox-2 + neuron in the striatum of a control monkey. Scale bar = 2 um. e1. Boxed area shows deposition of gold particles over the nucleus (arrowheads) close to a synapse (arrow), which identifies this cell as a neuron. Scale bar = 1 um. F. Sox-2 + astrocyte in the substantia nigra pars compacta. Scale bar = 10 µm. f1. Boxed area shows a dense bundle of intermediate filaments (arrows) that characterizes astrocytes. Arrowheads indicate gold particles in the nucleus. Scale bar = 1 µm. Abbreviations: subventricular zone: (SVZ); 3′, 3′-diaminobenzidine: (DAB).
    Figure Legend Snippet: Ultra-structural analysis of Sox-2 expression. A. Sox-2 immuno-gold label is observed in the nucleus (arrowheads) in ependymal cells and astrocytes in ultra-thin sections of the SVZ. Boxed areas show an ependymal cell (E) and an astrocyte (As). Ependymal layer (EL), gap layer (HL), astrocyte ribbon layer (R). Scale bar = 20 um; Scale bar in boxed areas = 1 um. a1. Magnification shows a Sox-2 + astrocyte and lack of immunoreactivity in an oligodendrocyte (O) and a microglial cell (Mi). Astrocytes are recognized by the presence of dense bundles of intermediate filaments (arrows). Scale bar = 2 um. B. Sox-2 immuno-gold staining in a group of neuroblasts (type A cells). Scale bar = 20 um. b1. Magnification of three neuroblasts showing the nuclear staining. Neuroblasts form chain-like structures with characteristic intercellular spaces (arrows). Scale bar = 2 um. C. Light microscopy image of a semi-thin section showing the distribution and characterization of cells expressing Sox-2 in the dentate gyrus. Sox-2 + cells are visualized by the nuclear dark brown DAB precipitate. The section was counterstained with toluidine blue. The line marks the boundary between the granular layer (GrL) and the hilus (H) where most labeled cells were located. Scale bar = 100 um. c1 ) Neurons in the granular layer were not Sox-2 + . Scale bar = 10 um; Insert = 1 um. D. Microphotograph of a striatal field of a control monkey showing a Sox-2 + astrocyte and unlabeled neuron (Neu) and microglial cells (Mi). Scale bar = 5 um. d1. Boxed area of the astrocyte (As) nucleus shows the gold particles (arrowheads). Scale bar = 1 um. E. A representative Sox-2 + neuron in the striatum of a control monkey. Scale bar = 2 um. e1. Boxed area shows deposition of gold particles over the nucleus (arrowheads) close to a synapse (arrow), which identifies this cell as a neuron. Scale bar = 1 um. F. Sox-2 + astrocyte in the substantia nigra pars compacta. Scale bar = 10 µm. f1. Boxed area shows a dense bundle of intermediate filaments (arrows) that characterizes astrocytes. Arrowheads indicate gold particles in the nucleus. Scale bar = 1 µm. Abbreviations: subventricular zone: (SVZ); 3′, 3′-diaminobenzidine: (DAB).

    Techniques Used: Expressing, Staining, Light Microscopy, Labeling

    9) Product Images from "CRE/CREB-Driven Up-Regulation of Gene Expression by Chronic Social Stress in CRE-Luciferase Transgenic Mice: Reversal by Antidepressant Treatment"

    Article Title: CRE/CREB-Driven Up-Regulation of Gene Expression by Chronic Social Stress in CRE-Luciferase Transgenic Mice: Reversal by Antidepressant Treatment

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0000431

    Reporter gene expression in transgenic CRE-Luc mice. Luciferase immunoreactivity in a CRE-Luc (A, C, D) and a control mouse (B). Luciferase antibody binding was visualized by 3,3′-diaminobenzidine staining on sagittal sections (brown reaction product in A to D); the section in D was counter stained with cresyl violet. Note that Luciferase immunoreactivity in the hippocampal formation (Hip) of the transgene mouse (A) is stronger than in the control (B). In the cerebellum (Cer in C), the molecular (mol) and the granule cell layer (gcl) as well the interposed nuclei (int) show moderate immunoreactivity. In the locus coeruleus (LC in D), distinct cells (brown) are moderately stained by the antibody. Abbreviations: cc, corpus callosum; CA1 and CA3, hippocampal regions cornu ammonis 1 and 3; DG, dentate gyrus; scp, superior cerebellar peduncle; Me5, mesencephalic trigeminal nucleus; me5, mesencephalic trigeminal tract; 8n, vestibulocochlear nerve. Scale bars: 500 µm (in A, B, C) and 200 µm (in D).
    Figure Legend Snippet: Reporter gene expression in transgenic CRE-Luc mice. Luciferase immunoreactivity in a CRE-Luc (A, C, D) and a control mouse (B). Luciferase antibody binding was visualized by 3,3′-diaminobenzidine staining on sagittal sections (brown reaction product in A to D); the section in D was counter stained with cresyl violet. Note that Luciferase immunoreactivity in the hippocampal formation (Hip) of the transgene mouse (A) is stronger than in the control (B). In the cerebellum (Cer in C), the molecular (mol) and the granule cell layer (gcl) as well the interposed nuclei (int) show moderate immunoreactivity. In the locus coeruleus (LC in D), distinct cells (brown) are moderately stained by the antibody. Abbreviations: cc, corpus callosum; CA1 and CA3, hippocampal regions cornu ammonis 1 and 3; DG, dentate gyrus; scp, superior cerebellar peduncle; Me5, mesencephalic trigeminal nucleus; me5, mesencephalic trigeminal tract; 8n, vestibulocochlear nerve. Scale bars: 500 µm (in A, B, C) and 200 µm (in D).

    Techniques Used: Expressing, Transgenic Assay, Mouse Assay, Luciferase, Binding Assay, Staining

    10) Product Images from "Thymic PTH Increases After Thyroparathyroidectomy in C57BL/KaLwRij Mice"

    Article Title: Thymic PTH Increases After Thyroparathyroidectomy in C57BL/KaLwRij Mice

    Journal: Endocrinology

    doi: 10.1210/en.2017-03083

    Thymus-derived PTH increases after TPTX. After serum PTH returned to a pre-TPTX level, TPTX and control mice were euthanized, and parathyroid and thyroid (Thyroid+Parathyroid), liver, and thymus tissues were removed. Harvested tissues were washed extensively in PBS and homogenized in T-PER buffer, and proteins were immunoprecipitated with goat anti-mouse PTH antibodies, including anti-mouse PTH(1-12) and anti-mouse PTH(53-84) or goat IgG as an isotype control. (A) PTH was detected in whole cell lysates (WCL) of the parathyroid+thyroid, after immunoprecipitation from the parathyroid+thyroid, and in the thymus (indicated by arrows). PTH protein was not detected in the liver. (B) Thymi from TPTX mice contained more PTH than thymi from nonoperated (control) mice. (C) Immunohistochemistry showing increased PTH protein production in the thymus after TPTX. Thymus sections were prepared and incubated with anti–PTH(1-12) antibody. The antibody was visualized with 3,3′-diaminobenzidine and counterstained with methyl green. The representative PTH signals are indicated by yellow arrows. (D) PTH-positive cells were enumerated in at least four sections from each group and plotted as mean ± standard error. Student t test was performed. *** P
    Figure Legend Snippet: Thymus-derived PTH increases after TPTX. After serum PTH returned to a pre-TPTX level, TPTX and control mice were euthanized, and parathyroid and thyroid (Thyroid+Parathyroid), liver, and thymus tissues were removed. Harvested tissues were washed extensively in PBS and homogenized in T-PER buffer, and proteins were immunoprecipitated with goat anti-mouse PTH antibodies, including anti-mouse PTH(1-12) and anti-mouse PTH(53-84) or goat IgG as an isotype control. (A) PTH was detected in whole cell lysates (WCL) of the parathyroid+thyroid, after immunoprecipitation from the parathyroid+thyroid, and in the thymus (indicated by arrows). PTH protein was not detected in the liver. (B) Thymi from TPTX mice contained more PTH than thymi from nonoperated (control) mice. (C) Immunohistochemistry showing increased PTH protein production in the thymus after TPTX. Thymus sections were prepared and incubated with anti–PTH(1-12) antibody. The antibody was visualized with 3,3′-diaminobenzidine and counterstained with methyl green. The representative PTH signals are indicated by yellow arrows. (D) PTH-positive cells were enumerated in at least four sections from each group and plotted as mean ± standard error. Student t test was performed. *** P

    Techniques Used: Derivative Assay, Mouse Assay, Immunoprecipitation, Immunohistochemistry, Incubation

    11) Product Images from "Differential Expression of IL-17RC Isoforms in Androgen-Dependent and Androgen-Independent Prostate Cancers 1"

    Article Title: Differential Expression of IL-17RC Isoforms in Androgen-Dependent and Androgen-Independent Prostate Cancers 1

    Journal:

    doi:

    Immunohistochemical staining of normal tissue microarrays. The slides were immunohistochemically stained for IL-17RC, using anti-ICD (left column) and anti-ECD (right column), together with ABC Elite kit and DAB substrate, and counterstained by hematoxylin.
    Figure Legend Snippet: Immunohistochemical staining of normal tissue microarrays. The slides were immunohistochemically stained for IL-17RC, using anti-ICD (left column) and anti-ECD (right column), together with ABC Elite kit and DAB substrate, and counterstained by hematoxylin.

    Techniques Used: Immunohistochemistry, Staining

    12) Product Images from "Connective Tissue Growth Factor Modulates Adult β-Cell Maturity and Proliferation to Promote β-Cell Regeneration in Mice"

    Article Title: Connective Tissue Growth Factor Modulates Adult β-Cell Maturity and Proliferation to Promote β-Cell Regeneration in Mice

    Journal: Diabetes

    doi: 10.2337/db14-1195

    Alterations in Tph1 expression and ERK1/2 signaling in response to CTGF and/or ablation. A – H : Representative images of islets at 2 days of CTGF. A and E : Control. B and F : CTGF. C and G : Ablation. D and H : Ablation+CTGF. Tph1: Primary antibodies were visualized via a DAB Peroxidase Substrate Kit (Vector Laboratories) and counterstained with hematoxylin. p-ERK1/2: Primary antibodies for p-ERK1/2 and insulin were visualized via a DAB Peroxidase Substrate Kit (Vector Laboratories) and an alkaline phosphatase Vector Blue Substrate Kit (Vector Laboratories), respectively. Brown and black arrowheads demark β-cells or other islet cells with activated ERK1/2 signaling, respectively. White arrowheads demark endothelial cells with activated ERK1/2 signaling. n = 4 for Tph1 and n = 3 for p-ERK1/2.
    Figure Legend Snippet: Alterations in Tph1 expression and ERK1/2 signaling in response to CTGF and/or ablation. A – H : Representative images of islets at 2 days of CTGF. A and E : Control. B and F : CTGF. C and G : Ablation. D and H : Ablation+CTGF. Tph1: Primary antibodies were visualized via a DAB Peroxidase Substrate Kit (Vector Laboratories) and counterstained with hematoxylin. p-ERK1/2: Primary antibodies for p-ERK1/2 and insulin were visualized via a DAB Peroxidase Substrate Kit (Vector Laboratories) and an alkaline phosphatase Vector Blue Substrate Kit (Vector Laboratories), respectively. Brown and black arrowheads demark β-cells or other islet cells with activated ERK1/2 signaling, respectively. White arrowheads demark endothelial cells with activated ERK1/2 signaling. n = 4 for Tph1 and n = 3 for p-ERK1/2.

    Techniques Used: Expressing, Plasmid Preparation

    13) Product Images from "High synovial expression of the inhibitory Fc?RIIb in rheumatoid arthritis"

    Article Title: High synovial expression of the inhibitory Fc?RIIb in rheumatoid arthritis

    Journal: Arthritis Research & Therapy

    doi: 10.1186/ar2206

    Expression of Fc gamma receptor (FcγR)IIb in healthy and rheumatoid arthritis (RA) synovia. (a) FcγRIIb was sparsely expressed in the few healthy synovial biopsies that were positively stained by GB3. The arrows indicate FcγRIIb positive cells in the sub-lining synovial layer and tissue. (b) The IgG1 isotype control antibody did not stain the healthy synovium. (c,e) Positive FcγRIIb expression was found in all the stained RA synovial biopsies. FcγRIIb staining was observed in the synovial lining and sub-lining layers, perivasculary and inside lymphocyte infiltrates (e). (d) No staining of the RA synovium was observed when the anti-FcγRIIb monoclonal antibody was bound by recombinant human soluble FcγRIIb prior to staining. (f) Synovial FcγRIIb expression was significantly up-regulated in RA patients ( n = 10) compared to healthy individuals ( n = 5) (mean values of two independent observers, SEM and SK with standard error of the mean, * p = 0.0104). DAB, diaminobenzidine. (Original magnification ×125.)
    Figure Legend Snippet: Expression of Fc gamma receptor (FcγR)IIb in healthy and rheumatoid arthritis (RA) synovia. (a) FcγRIIb was sparsely expressed in the few healthy synovial biopsies that were positively stained by GB3. The arrows indicate FcγRIIb positive cells in the sub-lining synovial layer and tissue. (b) The IgG1 isotype control antibody did not stain the healthy synovium. (c,e) Positive FcγRIIb expression was found in all the stained RA synovial biopsies. FcγRIIb staining was observed in the synovial lining and sub-lining layers, perivasculary and inside lymphocyte infiltrates (e). (d) No staining of the RA synovium was observed when the anti-FcγRIIb monoclonal antibody was bound by recombinant human soluble FcγRIIb prior to staining. (f) Synovial FcγRIIb expression was significantly up-regulated in RA patients ( n = 10) compared to healthy individuals ( n = 5) (mean values of two independent observers, SEM and SK with standard error of the mean, * p = 0.0104). DAB, diaminobenzidine. (Original magnification ×125.)

    Techniques Used: Expressing, Staining, Recombinant

    14) Product Images from "Antibodies to a Surface-Exposed, N-terminal Domain of Aggregation Substance Are Not Protective in the Rabbit Model of Enterococcus faecalis Infective Endocarditis"

    Article Title: Antibodies to a Surface-Exposed, N-terminal Domain of Aggregation Substance Are Not Protective in the Rabbit Model of Enterococcus faecalis Infective Endocarditis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.69.5.3305-3314.2001

    Vegetation formation on rabbit heart valve tissue by E. faecalis OG1SSp containing pINY1801 and demonstration of in vivo expression of Asc10. A typical vegetation formation on the rabbit aortic valve is shown in panel A. The arrowhead indicates the vegetation located within one of the aortic valves. Panels B and C (magnification, ×40) and D and E (magnification, ×200) represent consecutive 5-μm sections of a vegetation stained with preimmune serum (B and D) and AS 44–331 -immune serum (C and E). The secondary antibody was anti-rabbit horseradish peroxidase conjugate, and sections were developed with 3,3-diaminobenzidine as chromogen (brown color). Regions of the vegetation are identified as follows: a mass of E. faecalis cells (1) bordered by an influx of immune cells (2) and surrounded by a platelet/fibrin layer (3). Slides were counterstained with hematoxlyin.
    Figure Legend Snippet: Vegetation formation on rabbit heart valve tissue by E. faecalis OG1SSp containing pINY1801 and demonstration of in vivo expression of Asc10. A typical vegetation formation on the rabbit aortic valve is shown in panel A. The arrowhead indicates the vegetation located within one of the aortic valves. Panels B and C (magnification, ×40) and D and E (magnification, ×200) represent consecutive 5-μm sections of a vegetation stained with preimmune serum (B and D) and AS 44–331 -immune serum (C and E). The secondary antibody was anti-rabbit horseradish peroxidase conjugate, and sections were developed with 3,3-diaminobenzidine as chromogen (brown color). Regions of the vegetation are identified as follows: a mass of E. faecalis cells (1) bordered by an influx of immune cells (2) and surrounded by a platelet/fibrin layer (3). Slides were counterstained with hematoxlyin.

    Techniques Used: In Vivo, Expressing, Staining

    15) Product Images from "Heterogeneous induction of microglia M2a phenotype by central administration of interleukin-4"

    Article Title: Heterogeneous induction of microglia M2a phenotype by central administration of interleukin-4

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-014-0211-6

    Selective induction of Arg1 expression in a subset of microglia. Brains after 16 h intracerebroventricular treatment with vehicle (A , C) and IL4 (B , D) were formalin fixed and embedded in paraffin and analyzed by immunohistochemistry to visualize Iba1 (A , B) or Arg1 (C , D) in the frontal cortex. Arg1 expression is observed selectively in microglia-like cells after IL4 treatment and is restricted to a subset of cells (D) . Scale bar 100 μm. The results obtained by counting the number of Arg1-positive microglia cells and the total number of Iba1-positive microglia cells demonstrated that an ≈ 35% subset of microglia cells is Arg1 responder microglia (E) . Veh., vehicle.
    Figure Legend Snippet: Selective induction of Arg1 expression in a subset of microglia. Brains after 16 h intracerebroventricular treatment with vehicle (A , C) and IL4 (B , D) were formalin fixed and embedded in paraffin and analyzed by immunohistochemistry to visualize Iba1 (A , B) or Arg1 (C , D) in the frontal cortex. Arg1 expression is observed selectively in microglia-like cells after IL4 treatment and is restricted to a subset of cells (D) . Scale bar 100 μm. The results obtained by counting the number of Arg1-positive microglia cells and the total number of Iba1-positive microglia cells demonstrated that an ≈ 35% subset of microglia cells is Arg1 responder microglia (E) . Veh., vehicle.

    Techniques Used: Expressing, Immunohistochemistry

    Time course of IL4-induced M2 gene expression in frontal cortex and striatum . Following the indicated time intervals after intracerebroventricular injection of saline (Veh) or IL4, the RNA extracted from the frontal cortex (Fr cortex) and striatum was analyzed by real time PCR to evaluate (A) Fizz1 , (B) Arg1 , and (C) Ym1 gene expression. Data sets for each gene were calculated using the 2 -ddCt method with respect to the mean value of the 8 h vehicle group. Bars represent mean values ± SEM. * P
    Figure Legend Snippet: Time course of IL4-induced M2 gene expression in frontal cortex and striatum . Following the indicated time intervals after intracerebroventricular injection of saline (Veh) or IL4, the RNA extracted from the frontal cortex (Fr cortex) and striatum was analyzed by real time PCR to evaluate (A) Fizz1 , (B) Arg1 , and (C) Ym1 gene expression. Data sets for each gene were calculated using the 2 -ddCt method with respect to the mean value of the 8 h vehicle group. Bars represent mean values ± SEM. * P

    Techniques Used: Expressing, Injection, Real-time Polymerase Chain Reaction

    16) Product Images from "Midkine is a NF-?B-inducible gene that supports prostate cancer cell survival"

    Article Title: Midkine is a NF-?B-inducible gene that supports prostate cancer cell survival

    Journal: BMC Medical Genomics

    doi: 10.1186/1755-8794-1-6

    Immunohistochemical staining of prostate tissue microarrays. The early stage cancers were from radical prostatectomy specimens derived from patients with clinically localized prostate cancers; while the late stage cancers were derived from transurethral resection specimens of prostate cancers that had advanced beyond the stage treatable by radical prostatectomy; the normal prostate tissues were from the non-tumorous portions of the radical prostatectomy specimens; the prostate tissue microarray slides were stained with 0.6 μg/ml rabbit anti-midkine antibodies using the VECTSTAIN elite ABC Reagent and DAB Substrate Kit according to the manufacturer's protocol and counter-stained with hematoxylin; representative negative (Normal Epithelium and Early Stage Cancer) and strongly positive (Late Stage Cancer) midkine staining are shown; original magnification: × 400.
    Figure Legend Snippet: Immunohistochemical staining of prostate tissue microarrays. The early stage cancers were from radical prostatectomy specimens derived from patients with clinically localized prostate cancers; while the late stage cancers were derived from transurethral resection specimens of prostate cancers that had advanced beyond the stage treatable by radical prostatectomy; the normal prostate tissues were from the non-tumorous portions of the radical prostatectomy specimens; the prostate tissue microarray slides were stained with 0.6 μg/ml rabbit anti-midkine antibodies using the VECTSTAIN elite ABC Reagent and DAB Substrate Kit according to the manufacturer's protocol and counter-stained with hematoxylin; representative negative (Normal Epithelium and Early Stage Cancer) and strongly positive (Late Stage Cancer) midkine staining are shown; original magnification: × 400.

    Techniques Used: Immunohistochemistry, Staining, Derivative Assay, Microarray

    17) Product Images from "Calretinin and calbindin architecture of the midline thalamus associated with prefrontal-hippocampal circuitry"

    Article Title: Calretinin and calbindin architecture of the midline thalamus associated with prefrontal-hippocampal circuitry

    Journal: bioRxiv

    doi: 10.1101/2020.07.21.214973

    Distribution of DAB CR + and DAB CB + cells is not the same across all RE internal subdivisions A: Brightfield images showing the distribution of DAB CR + cells in RE across the rostro-caudal axis of the thalamus. CR + cell area density varied depending on the subdivision of RE in which they were located. B: Distribution of DAB CB + cells. When compared, CR + and CB + cell distribution in all RE’s subregions and across the rostral to caudal levels did not appear to be the same. Overlay shown adapted from Swanson (2018) to highlight all RE internal subdivisions. Scale bar = 100μm. C: Comparison of DAB CR + and DAB CB + cell area density (cells/0.01mm 2 ) in all subdivisions of RE across the rostro-caudal axis. CB + cell densities were higher than CR + cell densities (except REm). Additionally, REl, REv and REcd subdivisions exhibited large CB + cell densities compared to other RE subregions. A moderate size effect was found for CB + cell area density across all levels and RE subdivisions (Hedges’ d=0.32). Abbreviations: β, bregma; CB, calbindin; CR, calretinin; DAB, 3,3’-Diaminobenzidine; PRe, perireuniens, RE, nucleus reuniens of the thalamus, REa, reuniens rostral division anterior part; REd, reuniens rostral division dorsal part; REl, reuniens rostral division lateral part; REm, reuniens rostral division median part; REv, reuniens rostral division ventral part; REcm, reuniens caudal division median part; REcd, reuniens caudal division dorsal part; REcp, reuniens caudal division posterior part.
    Figure Legend Snippet: Distribution of DAB CR + and DAB CB + cells is not the same across all RE internal subdivisions A: Brightfield images showing the distribution of DAB CR + cells in RE across the rostro-caudal axis of the thalamus. CR + cell area density varied depending on the subdivision of RE in which they were located. B: Distribution of DAB CB + cells. When compared, CR + and CB + cell distribution in all RE’s subregions and across the rostral to caudal levels did not appear to be the same. Overlay shown adapted from Swanson (2018) to highlight all RE internal subdivisions. Scale bar = 100μm. C: Comparison of DAB CR + and DAB CB + cell area density (cells/0.01mm 2 ) in all subdivisions of RE across the rostro-caudal axis. CB + cell densities were higher than CR + cell densities (except REm). Additionally, REl, REv and REcd subdivisions exhibited large CB + cell densities compared to other RE subregions. A moderate size effect was found for CB + cell area density across all levels and RE subdivisions (Hedges’ d=0.32). Abbreviations: β, bregma; CB, calbindin; CR, calretinin; DAB, 3,3’-Diaminobenzidine; PRe, perireuniens, RE, nucleus reuniens of the thalamus, REa, reuniens rostral division anterior part; REd, reuniens rostral division dorsal part; REl, reuniens rostral division lateral part; REm, reuniens rostral division median part; REv, reuniens rostral division ventral part; REcm, reuniens caudal division median part; REcd, reuniens caudal division dorsal part; REcp, reuniens caudal division posterior part.

    Techniques Used:

    18) Product Images from "Rapid in vitro assays for screening neutralizing antibodies and antivirals against SARS-CoV-2"

    Article Title: Rapid in vitro assays for screening neutralizing antibodies and antivirals against SARS-CoV-2

    Journal: bioRxiv

    doi: 10.1101/2020.07.22.216648

    PRMNT assay to identify SARS-CoV-2 NAbs, peroxidase staining. A) Pre-treatment: 2-fold serially diluted mAbs, pAbs, or serum samples are pre-incubated with ~100-200 PFU of SARS-CoV-2 for 1 h at RT. After 1 h pre-incubation, confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with the mAb/pAb/serum-virus mixture for 1 h at RT. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. After 1 h of virus adsorption, cells are washed 3X with PBS and post-infection media containing 1% Avicel is added in all the wells. B) Post-infection: confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU/well of SARS-CoV-2. After 1 h of virus adsorption, the infection media is replaced with post-infection media containing 1% Avicel and 2-fold serially diluted mAbs, pAbs or serum samples. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. A and B) At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD anti-mouse Ab (diluted according to the manufacturer’s instruction) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit. Positive staining plaques in each of the wells are quantified using am ELISPOT plate reader. The NT 50 is calculated as the highest dilution of the mAb, pAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve. Dotted line indicates 50% neutralization. Data were expressed as mean and SD from triplicate wells.
    Figure Legend Snippet: PRMNT assay to identify SARS-CoV-2 NAbs, peroxidase staining. A) Pre-treatment: 2-fold serially diluted mAbs, pAbs, or serum samples are pre-incubated with ~100-200 PFU of SARS-CoV-2 for 1 h at RT. After 1 h pre-incubation, confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with the mAb/pAb/serum-virus mixture for 1 h at RT. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. After 1 h of virus adsorption, cells are washed 3X with PBS and post-infection media containing 1% Avicel is added in all the wells. B) Post-infection: confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU/well of SARS-CoV-2. After 1 h of virus adsorption, the infection media is replaced with post-infection media containing 1% Avicel and 2-fold serially diluted mAbs, pAbs or serum samples. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. A and B) At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD anti-mouse Ab (diluted according to the manufacturer’s instruction) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit. Positive staining plaques in each of the wells are quantified using am ELISPOT plate reader. The NT 50 is calculated as the highest dilution of the mAb, pAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve. Dotted line indicates 50% neutralization. Data were expressed as mean and SD from triplicate wells.

    Techniques Used: Staining, Incubation, Infection, Adsorption, Enzyme-linked Immunospot, Neutralization

    A PRMNT assay to identify SARS-CoV-2 antivirals, peroxidase staining: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU/well of SARS-CoV-2. After 1 h of virus adsorption, media is replaced with fresh infection media containing 1% Avicel and 2-fold serially diluted drugs. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD anti-mouse Ab (diluted according to the manufacturer’s instruction) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit. Positive staining plaques in each of the wells are quantified using an ELISPOT plate reader. The effective concentration 50 (EC 50 ) is calculated as the highest dilution of the mAb, pAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve. Dotted line indicates 50% neutralization. Data were expressed as mean and SD from triplicate wells.
    Figure Legend Snippet: A PRMNT assay to identify SARS-CoV-2 antivirals, peroxidase staining: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU/well of SARS-CoV-2. After 1 h of virus adsorption, media is replaced with fresh infection media containing 1% Avicel and 2-fold serially diluted drugs. Cells in row 11 are incubated with virus only and cells in row are mock-infected and are used as internal controls in each of the plates. At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD anti-mouse Ab (diluted according to the manufacturer’s instruction) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit. Positive staining plaques in each of the wells are quantified using an ELISPOT plate reader. The effective concentration 50 (EC 50 ) is calculated as the highest dilution of the mAb, pAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve. Dotted line indicates 50% neutralization. Data were expressed as mean and SD from triplicate wells.

    Techniques Used: Staining, Infection, Adsorption, Incubation, Enzyme-linked Immunospot, Concentration Assay, Neutralization

    Schematic representation of the PRMNT assay to identify SARS-CoV-2 antivirals: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU of SARS-CoV-2 for 1 h at 37°C. Cells in rows 11 and 12 are incubated with virus only and mock-infected, respectively, and are used as internal controls in each of the plates. After 1 h viral absorption, cells are incubated with post-infection media containing 2-fold serial dilutions of the antivirals containing 1% Avicel. At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1 μg/mL of the SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD ( Fig. 5 ) or with IRDye 800CW goat anti-mouse IgG secondary Ab, and DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 6 ) at 37°C. After 30 minutes incubation with the secondary POD Ab, cells are washed 3X with PBS and developed with the DAB substrate kit ( Fig. 5 ). Cells stained with the IRDye 800CW goat anti-mouse IgG secondary Ab are simultaneously incubated with DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 6 ). Positive stained cells in each of the wells of the 96-well plate are quantified using an ELISPOT plate reader ( Fig. 5 ) or in the Odyssey Sa Infrared Imaging System ( Fig. 6 ). The effective concentration 50 (EC 50 ) is calculated as the highest dilution of the drug that prevents 50% plaque formation in infected cells, determined by a sigmo idal dose response curve ( Figs. 5 and 6 ).
    Figure Legend Snippet: Schematic representation of the PRMNT assay to identify SARS-CoV-2 antivirals: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU of SARS-CoV-2 for 1 h at 37°C. Cells in rows 11 and 12 are incubated with virus only and mock-infected, respectively, and are used as internal controls in each of the plates. After 1 h viral absorption, cells are incubated with post-infection media containing 2-fold serial dilutions of the antivirals containing 1% Avicel. At 24 h p.i., cells are fixed with 10% formalin solution. After 24 h fixation, cells are washed 3X with PBS and incubated with 1 μg/mL of the SARS-CoV-1 cross-reactive NP mAb (1C7) at 37°C. After 1 h incubation with the primary mAb, cells are washed 3X with PBS and incubated with a secondary POD ( Fig. 5 ) or with IRDye 800CW goat anti-mouse IgG secondary Ab, and DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 6 ) at 37°C. After 30 minutes incubation with the secondary POD Ab, cells are washed 3X with PBS and developed with the DAB substrate kit ( Fig. 5 ). Cells stained with the IRDye 800CW goat anti-mouse IgG secondary Ab are simultaneously incubated with DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 6 ). Positive stained cells in each of the wells of the 96-well plate are quantified using an ELISPOT plate reader ( Fig. 5 ) or in the Odyssey Sa Infrared Imaging System ( Fig. 6 ). The effective concentration 50 (EC 50 ) is calculated as the highest dilution of the drug that prevents 50% plaque formation in infected cells, determined by a sigmo idal dose response curve ( Figs. 5 and 6 ).

    Techniques Used: Infection, Incubation, Staining, Enzyme-linked Immunospot, Imaging, Concentration Assay

    Schematic representation of the PRMNT assay to identify SARS-CoV-2 NAbs: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU of SARS-CoV-2 pre-incubated (A) or post-treated (B) with a 2-fold serially diluted mAbs, pAbs, and/or serum samples. Cells in rows 11 and 12 are incubated as virus only and mock only infected cells, respectively, and are used as internal controls in the assay. After 1 h absorption of the virus-mAb mixture (A) or virus alone (B), infected cells are washed 3X with PBS and post-infection media containing 1% Avicel is added in all the wells alone (A) or with the 2-fold serially diluted mAbs and/or serum samples (B). At 24 h p.i., cells in the 96-well plate are fixed with 10% formalin solution. After 24 h, fixed cells are washed 3X with PBS and incubated with 1 μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7, 1 μg/mL) at 37°C. After 1 h incubation with the primary Ab, cells are washed 3X with PBS and incubated with a secondary POD ( Fig. 2 ) or IRDye 800CW goat anti-mouse IgG secondary Ab ( Fig. 3 ) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit ( Fig. 2 ). Cells stained with the IRDye 800CW goat anti-mouse IgG secondary Ab are simultaneously incubated with DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 3 ). Positive staining plaques in each of the wells of the 96-well plate is quantified using an ELISPOT plate reader ( Fig. 2 ) or an Odyssey Sa Infrared Imaging System ( Fig. 3 ). The neutralizing titer 50 (NT 50 ) is calculated as the highest dilution of the mAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve ( Figs. 2 and 3 ).
    Figure Legend Snippet: Schematic representation of the PRMNT assay to identify SARS-CoV-2 NAbs: Confluent monolayers of Vero E6 cells (96-well plate format, ~4 x 10 4 cells/well, triplicates) are infected with ~100-200 PFU of SARS-CoV-2 pre-incubated (A) or post-treated (B) with a 2-fold serially diluted mAbs, pAbs, and/or serum samples. Cells in rows 11 and 12 are incubated as virus only and mock only infected cells, respectively, and are used as internal controls in the assay. After 1 h absorption of the virus-mAb mixture (A) or virus alone (B), infected cells are washed 3X with PBS and post-infection media containing 1% Avicel is added in all the wells alone (A) or with the 2-fold serially diluted mAbs and/or serum samples (B). At 24 h p.i., cells in the 96-well plate are fixed with 10% formalin solution. After 24 h, fixed cells are washed 3X with PBS and incubated with 1 μg/mL of a SARS-CoV-1 cross-reactive NP mAb (1C7, 1 μg/mL) at 37°C. After 1 h incubation with the primary Ab, cells are washed 3X with PBS and incubated with a secondary POD ( Fig. 2 ) or IRDye 800CW goat anti-mouse IgG secondary Ab ( Fig. 3 ) at 37°C. After 30 minutes incubation with the secondary Ab, cells are washed 3X with PBS and developed with the DAB substrate kit ( Fig. 2 ). Cells stained with the IRDye 800CW goat anti-mouse IgG secondary Ab are simultaneously incubated with DRAQ5 ™ Fluorescent Probe Solution for nuclear staining ( Fig. 3 ). Positive staining plaques in each of the wells of the 96-well plate is quantified using an ELISPOT plate reader ( Fig. 2 ) or an Odyssey Sa Infrared Imaging System ( Fig. 3 ). The neutralizing titer 50 (NT 50 ) is calculated as the highest dilution of the mAb or sera that prevents 50% plaque formation in infected cells, determined by a sigmoidal dose response curve ( Figs. 2 and 3 ).

    Techniques Used: Infection, Incubation, Staining, Enzyme-linked Immunospot, Imaging

    19) Product Images from "Overexpression of DeltaFosB in nucleus accumbens mimics the protective addiction phenotype, but not the protective depression phenotype of environmental enrichment"

    Article Title: Overexpression of DeltaFosB in nucleus accumbens mimics the protective addiction phenotype, but not the protective depression phenotype of environmental enrichment

    Journal: Frontiers in Behavioral Neuroscience

    doi: 10.3389/fnbeh.2014.00297

    Stress and ∆FosB in EC and IC rats. (A–D) Representative immunohistochemistry DAB staining of ΔFosB in NAc shell and core of IC ( A and B ) and EC ( C and D ) rats with ( B and D ) and without ( A and C ) repeated stress ( N = 3). (E) Quantification of the number of ΔFosB positive cells (±SEM) in NAc shell induced by repeated restraint stress in IC and EC rats. (F) Number of ΔFosB positive cells (±SEM) in NAc core induced by repeated restraint stress. (G) Fold change of ΔFosB mRNA (±SEM) induced by acute and repeated restraint stress in IC and EC rats (30 min; N = 5). Asterisks (*) denote significant difference from corresponding control.
    Figure Legend Snippet: Stress and ∆FosB in EC and IC rats. (A–D) Representative immunohistochemistry DAB staining of ΔFosB in NAc shell and core of IC ( A and B ) and EC ( C and D ) rats with ( B and D ) and without ( A and C ) repeated stress ( N = 3). (E) Quantification of the number of ΔFosB positive cells (±SEM) in NAc shell induced by repeated restraint stress in IC and EC rats. (F) Number of ΔFosB positive cells (±SEM) in NAc core induced by repeated restraint stress. (G) Fold change of ΔFosB mRNA (±SEM) induced by acute and repeated restraint stress in IC and EC rats (30 min; N = 5). Asterisks (*) denote significant difference from corresponding control.

    Techniques Used: Immunohistochemistry, Staining

    20) Product Images from "Extracellular Administration of BCL2 Protein Reduces Apoptosis and Improves Survival in a Murine Model of Sepsis"

    Article Title: Extracellular Administration of BCL2 Protein Reduces Apoptosis and Improves Survival in a Murine Model of Sepsis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0014729

    Treatment with rhBCL2A1 increases expression of endogenous BCL2 following CLP. Mice were treated with a single i.p. injection of 1 µg rhBim (A, C) or rhBCL2A1 (B, D) at 18 hours prior to CLP. Heart (A, B) and intestine (C, D) were examined at 24 hours following CLP. Endogenous mouse BCL2 protein was detected by immuno-staining with rabbit anti-mouse BCL2 antibody detected with 3, 3′-diaminobenzidine (brown) and the nuclei were stained with hematoxylin (purple). Representative FACS analysis of endogenous BCL2 expression in cardiomyocytes from mice treated as above with rhBim (E) or rhBCL2A1 (F). Green line represents isotype control and red line represents anti-mouse BCL2 antibody.
    Figure Legend Snippet: Treatment with rhBCL2A1 increases expression of endogenous BCL2 following CLP. Mice were treated with a single i.p. injection of 1 µg rhBim (A, C) or rhBCL2A1 (B, D) at 18 hours prior to CLP. Heart (A, B) and intestine (C, D) were examined at 24 hours following CLP. Endogenous mouse BCL2 protein was detected by immuno-staining with rabbit anti-mouse BCL2 antibody detected with 3, 3′-diaminobenzidine (brown) and the nuclei were stained with hematoxylin (purple). Representative FACS analysis of endogenous BCL2 expression in cardiomyocytes from mice treated as above with rhBim (E) or rhBCL2A1 (F). Green line represents isotype control and red line represents anti-mouse BCL2 antibody.

    Techniques Used: Expressing, Mouse Assay, Injection, Immunostaining, Staining, FACS

    Treatment with rhBCL2 protein reduces cleaved caspase-3 in heart and intestine of mice following CLP. Mice were treated with a single i.p. injection of 1 µg rhBim (A, C) or 1 µg rhBCL2 (B, D) at 18 hours prior to CLP. Heart (A, B) or intestine (C, D) was stained for cleaved caspase with 3, 3′-diaminobenzidine (brown). Nuclei were stained with hematoxylin (purple). Original magnification is 400×. Panels E and F show quantitation of cleaved caspase-3 expression in myocardium (E) and intestinal villi (F) in mice treated with rhBim compared to those treated with rhBCL2. *p
    Figure Legend Snippet: Treatment with rhBCL2 protein reduces cleaved caspase-3 in heart and intestine of mice following CLP. Mice were treated with a single i.p. injection of 1 µg rhBim (A, C) or 1 µg rhBCL2 (B, D) at 18 hours prior to CLP. Heart (A, B) or intestine (C, D) was stained for cleaved caspase with 3, 3′-diaminobenzidine (brown). Nuclei were stained with hematoxylin (purple). Original magnification is 400×. Panels E and F show quantitation of cleaved caspase-3 expression in myocardium (E) and intestinal villi (F) in mice treated with rhBim compared to those treated with rhBCL2. *p

    Techniques Used: Mouse Assay, Injection, Staining, Quantitation Assay, Expressing

    21) Product Images from "Neurons of self-defence: neuronal innervation of the exocrine defence glands in stick insects"

    Article Title: Neurons of self-defence: neuronal innervation of the exocrine defence glands in stick insects

    Journal: Frontiers in Zoology

    doi: 10.1186/s12983-015-0122-0

    Tracing preparations from S. sipylus with neurons innervating the defence gland via the intersegmental nerve complex ( N. posterior SOG and N. anterior T1) in a the suboesophageal ganglion and ( b – d ) the prothoracic ganglion. In the preparations in dorsal view ( a – b , d ), the left nerves were filled. Prothoracic dorsal neurons located posterior in the ganglion are DUM neurons with bilaterally symmetrical neurites, see arrows in ( b , d ). c Lateral view of the prothoracic ganglion reveals two DUM neuron somata in the dorsal ganglion in close contact. A further prothoracic neuron occurs located at the dorsal midline ( d ). The neurite runs medially ( white arrowhead ) in the ganglion. Nerves in the ganglia were traced using neurobiotin solution. Abbreviations: SOG, suboesophageal ganglion; T1, prothoracic ganglion. Scale bars: 100 μm
    Figure Legend Snippet: Tracing preparations from S. sipylus with neurons innervating the defence gland via the intersegmental nerve complex ( N. posterior SOG and N. anterior T1) in a the suboesophageal ganglion and ( b – d ) the prothoracic ganglion. In the preparations in dorsal view ( a – b , d ), the left nerves were filled. Prothoracic dorsal neurons located posterior in the ganglion are DUM neurons with bilaterally symmetrical neurites, see arrows in ( b , d ). c Lateral view of the prothoracic ganglion reveals two DUM neuron somata in the dorsal ganglion in close contact. A further prothoracic neuron occurs located at the dorsal midline ( d ). The neurite runs medially ( white arrowhead ) in the ganglion. Nerves in the ganglia were traced using neurobiotin solution. Abbreviations: SOG, suboesophageal ganglion; T1, prothoracic ganglion. Scale bars: 100 μm

    Techniques Used:

    a Lateral view on the suboesophageal ganglion of P. schultei showing the ILN and VMNs. Dotted circle outlines the dorsal area of neurites. b i-iii The number of VMNs is variable, as shown with 2 ( b i ), 3 ( b ii ) or 4 ( b iii ) neurons. Preparations of E. tiaratum from females. The nerves in all original ganglia preparations were traced using neurobiotin solution as tracer. Abbreviations: ILN, ipsilateral neuron; VMN, ventral median neurons. Scale bars: ( a ), 100 μm; ( b i – iii ), 50 μm
    Figure Legend Snippet: a Lateral view on the suboesophageal ganglion of P. schultei showing the ILN and VMNs. Dotted circle outlines the dorsal area of neurites. b i-iii The number of VMNs is variable, as shown with 2 ( b i ), 3 ( b ii ) or 4 ( b iii ) neurons. Preparations of E. tiaratum from females. The nerves in all original ganglia preparations were traced using neurobiotin solution as tracer. Abbreviations: ILN, ipsilateral neuron; VMN, ventral median neurons. Scale bars: ( a ), 100 μm; ( b i – iii ), 50 μm

    Techniques Used:

    Distribution of neurons in the suboesophageal ( top row ) and prothoracic ganglion ( bottom row ) revealed by retrograde tracing of the left N. anterior SOG in a P. schultei , b S. sipylus , c C. morosus , and d E. tiaratum . Ganglia are shown in dorsal (top) view after filling of the left N. anterior SOG. The nerves in all original ganglia preparations were traced using neurobiotin solution as tracer. Abbreviations: CLN, contralateral neuron; ILN, ispilateral neuron; N. a., Nervus anterior SOG; PIN, prothoracic neuron; VMN, ventral median neurons. All scale bars: 100 μm
    Figure Legend Snippet: Distribution of neurons in the suboesophageal ( top row ) and prothoracic ganglion ( bottom row ) revealed by retrograde tracing of the left N. anterior SOG in a P. schultei , b S. sipylus , c C. morosus , and d E. tiaratum . Ganglia are shown in dorsal (top) view after filling of the left N. anterior SOG. The nerves in all original ganglia preparations were traced using neurobiotin solution as tracer. Abbreviations: CLN, contralateral neuron; ILN, ispilateral neuron; N. a., Nervus anterior SOG; PIN, prothoracic neuron; VMN, ventral median neurons. All scale bars: 100 μm

    Techniques Used: Retrograde Tracing

    Innervation of the defence gland by different nerves in C. morosus ( a , b ) and S. sipylus ( c – f ). In C. morosus , tracing of N. anterior SOG reveals the innervation of the defence gland and also adjacent muscles. Also visible is the nerve branch from the N. posterior SOG/ N. anterior T1 which is not stained ( empty arrow ). b Tracing of N. transversus does not result in staining of nerve fibres on the gland but of a small muscle ( asterisk ) next to the gland by a thin nerve branch ( black arrow ). The nerve branch from N. transversus to the gland is not stained ( empty arrow ). Scale bars: 500 μm. In S. sipylus , the gland innervation shown by neuronal branches on the gland surface from several nerves: c the N. anterior SOG, d the N. anterior SOG via the prothoracic-suboesophageal connective, e the N. posterior SOG, and d the N. anterior T1. In all preparations neurobiotin solution was used as tracer. Scale bars: 500 μm
    Figure Legend Snippet: Innervation of the defence gland by different nerves in C. morosus ( a , b ) and S. sipylus ( c – f ). In C. morosus , tracing of N. anterior SOG reveals the innervation of the defence gland and also adjacent muscles. Also visible is the nerve branch from the N. posterior SOG/ N. anterior T1 which is not stained ( empty arrow ). b Tracing of N. transversus does not result in staining of nerve fibres on the gland but of a small muscle ( asterisk ) next to the gland by a thin nerve branch ( black arrow ). The nerve branch from N. transversus to the gland is not stained ( empty arrow ). Scale bars: 500 μm. In S. sipylus , the gland innervation shown by neuronal branches on the gland surface from several nerves: c the N. anterior SOG, d the N. anterior SOG via the prothoracic-suboesophageal connective, e the N. posterior SOG, and d the N. anterior T1. In all preparations neurobiotin solution was used as tracer. Scale bars: 500 μm

    Techniques Used: Staining

    Backfill preparations with neurons supplying the N. anterior SOG in the suboesophageal and prothoracic ganglia. The filled nerve N. anterior SOG is indicated by asterisks. Stained fibres in the connectives and peripheral nerves of P. schultei are indicated by arrowheads. Preparations of a i-ii P. schultei are from a male individual, and of b E. tiaratum from a female, and c C. morosus from a female. In all preparations neurobiotin solution was used as tracer. Abbreviations: CLN, contralateral neurons; ILN, ipsilateral neuron; PIN, prothoracic intersegmental neuron; SOG, suboesophageal ganglion; T1, prothoracic ganglion; VMN, ventral medial neurons. All scale bars: 100 μm
    Figure Legend Snippet: Backfill preparations with neurons supplying the N. anterior SOG in the suboesophageal and prothoracic ganglia. The filled nerve N. anterior SOG is indicated by asterisks. Stained fibres in the connectives and peripheral nerves of P. schultei are indicated by arrowheads. Preparations of a i-ii P. schultei are from a male individual, and of b E. tiaratum from a female, and c C. morosus from a female. In all preparations neurobiotin solution was used as tracer. Abbreviations: CLN, contralateral neurons; ILN, ipsilateral neuron; PIN, prothoracic intersegmental neuron; SOG, suboesophageal ganglion; T1, prothoracic ganglion; VMN, ventral medial neurons. All scale bars: 100 μm

    Techniques Used: Staining

    22) Product Images from "Intravenous immunoglobulin prevents experimental autoimmune myositis in SJL mice by reducing anti-myosin antibody and by blocking complement deposition"

    Article Title: Intravenous immunoglobulin prevents experimental autoimmune myositis in SJL mice by reducing anti-myosin antibody and by blocking complement deposition

    Journal: Clinical and Experimental Immunology

    doi: 10.1046/j.1365-2249.2001.01499.x

    Blockade of mouse IgG and complement C3 deposition in muscle fibres of EAM mice by IVIG. SJL mice were injected with MB/CFA on day 0 and 21. IVIG (400 mg/kg/day) was administrated i.v. for 5 consecutive days from day 21. After 6, 8, 10 and 12 weeks from the first immunization day tissue specimens from thigh muscles were prepared. Frozen sections were prepared and immunohistochemical localization of mouse IgG (HRP-labelled antimoue IgG antibody) and mouse C3 (goat IgG antimouse C3 antibody and HRP-labelled antigoat IgG) on the sections was detected using a DAB substrate kit. According to the intensity of labelling, immunoreaction was represented and scored as follows; –: negative (score 0), ± : very weak (0·5), + : weak (1), + + : moderate (2), + + + : strong (3). Values are expressed as the mean ±SE ( n = 4–5). ○ control; • IVIG.
    Figure Legend Snippet: Blockade of mouse IgG and complement C3 deposition in muscle fibres of EAM mice by IVIG. SJL mice were injected with MB/CFA on day 0 and 21. IVIG (400 mg/kg/day) was administrated i.v. for 5 consecutive days from day 21. After 6, 8, 10 and 12 weeks from the first immunization day tissue specimens from thigh muscles were prepared. Frozen sections were prepared and immunohistochemical localization of mouse IgG (HRP-labelled antimoue IgG antibody) and mouse C3 (goat IgG antimouse C3 antibody and HRP-labelled antigoat IgG) on the sections was detected using a DAB substrate kit. According to the intensity of labelling, immunoreaction was represented and scored as follows; –: negative (score 0), ± : very weak (0·5), + : weak (1), + + : moderate (2), + + + : strong (3). Values are expressed as the mean ±SE ( n = 4–5). ○ control; • IVIG.

    Techniques Used: Mouse Assay, Injection, Immunohistochemistry

    23) Product Images from "Impaired wound healing in transgenic mice overexpressing the activin antagonist follistatin in the epidermis"

    Article Title: Impaired wound healing in transgenic mice overexpressing the activin antagonist follistatin in the epidermis

    Journal: The EMBO Journal

    doi: 10.1093/emboj/20.19.5361

    Fig. 4. Normal epidermal architecture and keratinocyte proliferation in the follistatin-transgenic mice. ( A ) Paraffin sections (6 µm) from the tail were stained using the Masson trichrome protocol. D, dermis; E, epidermis; F, fatty tissue; HF, hair follicle. ( B ) Detection of proliferating cells in the tail skin of control mice and transgenic mice. Paraffin sections of the tail skin of BrdU-injected control mice (left panel) and transgenic mice (right panel) were incubated with a peroxidase-conjugated anti-BrdU antibody, stained with the diaminobenzidine peroxidase-staining kit, and counterstained with hematoxylin. Stained nuclei are indicated by arrows. E, epidermis; D, dermis; HF, hair follicle. Bar indicates 50 µm in (A) and 30 µm in (B).
    Figure Legend Snippet: Fig. 4. Normal epidermal architecture and keratinocyte proliferation in the follistatin-transgenic mice. ( A ) Paraffin sections (6 µm) from the tail were stained using the Masson trichrome protocol. D, dermis; E, epidermis; F, fatty tissue; HF, hair follicle. ( B ) Detection of proliferating cells in the tail skin of control mice and transgenic mice. Paraffin sections of the tail skin of BrdU-injected control mice (left panel) and transgenic mice (right panel) were incubated with a peroxidase-conjugated anti-BrdU antibody, stained with the diaminobenzidine peroxidase-staining kit, and counterstained with hematoxylin. Stained nuclei are indicated by arrows. E, epidermis; D, dermis; HF, hair follicle. Bar indicates 50 µm in (A) and 30 µm in (B).

    Techniques Used: Transgenic Assay, Mouse Assay, Staining, Injection, Incubation

    24) Product Images from "Loss of Hif-2α Rescues the Hif-1α Deletion Phenotype of Neonatal Respiratory Distress In Mice"

    Article Title: Loss of Hif-2α Rescues the Hif-1α Deletion Phenotype of Neonatal Respiratory Distress In Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0139270

    Survivability Plot. Viability of neonatal Hif-1αΔ/Δ , Hif-2αΔ/Δ and Hif-1/2αΔ/Δ pups. ( A ) All Hif-1αΔ/Δ pups showed signs of respiratory distress whereas all Hif-2αΔ/Δ pups were viable with no obvious phenotype. Simultaneous removal of both Hifα forms resulted in approximately 79% survival. ( B ) Time-course of various Hif-1α mice with doxycycline (DOXY) exposure
    Figure Legend Snippet: Survivability Plot. Viability of neonatal Hif-1αΔ/Δ , Hif-2αΔ/Δ and Hif-1/2αΔ/Δ pups. ( A ) All Hif-1αΔ/Δ pups showed signs of respiratory distress whereas all Hif-2αΔ/Δ pups were viable with no obvious phenotype. Simultaneous removal of both Hifα forms resulted in approximately 79% survival. ( B ) Time-course of various Hif-1α mice with doxycycline (DOXY) exposure

    Techniques Used: Mouse Assay

    Merged Transcriptional Network with c-Myc . The linkage between the Hif-1α transcriptional network and c-Myc were merged and common genes in both networks were identified. Hif-1α -specific (blue circles), c-Myc -specific (green circles), and common genes (half blue, half green circles) are separated by function.
    Figure Legend Snippet: Merged Transcriptional Network with c-Myc . The linkage between the Hif-1α transcriptional network and c-Myc were merged and common genes in both networks were identified. Hif-1α -specific (blue circles), c-Myc -specific (green circles), and common genes (half blue, half green circles) are separated by function.

    Techniques Used:

    Merged Transcriptional Network with C/ebpα . The linkage between the Hif-1α transcriptional network and C/ebpα were merged and common genes in both networks were identified. Hif-1α -specific (green circles), C/ebpα -specific (blue circles), and common genes (half blue, half green circles) are separated by function.
    Figure Legend Snippet: Merged Transcriptional Network with C/ebpα . The linkage between the Hif-1α transcriptional network and C/ebpα were merged and common genes in both networks were identified. Hif-1α -specific (green circles), C/ebpα -specific (blue circles), and common genes (half blue, half green circles) are separated by function.

    Techniques Used:

    Genotyping of transgenes. PCR genotyping was performed for all four transgenes as described in materials and methods. Sizes of the amplified products obtained are: 240 bp for Hif-1α (wild type); 274 bp for Hif-1α flox/flox ; 410 bp for Hif-2α (wild type); 444bp for Hif-2α flox/flox ; 370 bpfor Cre transgene; 350 bpfor rtTA transgene. One representative sample was genotyped for the four transgenes from each of three generated mouse strains: SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-1α fl/fl ( Lane 1 ), SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-2α fl/fl mouse ( Lane 2 ), and SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-1α/2α fl/fl ( Lane 3 )).
    Figure Legend Snippet: Genotyping of transgenes. PCR genotyping was performed for all four transgenes as described in materials and methods. Sizes of the amplified products obtained are: 240 bp for Hif-1α (wild type); 274 bp for Hif-1α flox/flox ; 410 bp for Hif-2α (wild type); 444bp for Hif-2α flox/flox ; 370 bpfor Cre transgene; 350 bpfor rtTA transgene. One representative sample was genotyped for the four transgenes from each of three generated mouse strains: SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-1α fl/fl ( Lane 1 ), SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-2α fl/fl mouse ( Lane 2 ), and SPC-rtTA -/tg /(tetO) 7 -Cre -/tg /Hif-1α/2α fl/fl ( Lane 3 )).

    Techniques Used: Polymerase Chain Reaction, Amplification, Generated

    25) Product Images from "White matter involvement in sporadic Creutzfeldt-Jakob disease"

    Article Title: White matter involvement in sporadic Creutzfeldt-Jakob disease

    Journal: Brain

    doi: 10.1093/brain/awu298

    Histopathology findings. Examples of the main neuropathological changes found in sporadic CJD cases without white matter PrP Sc ( A–F , Subject 18, see Table 3 ) and with mild white matter PrP Sc ( G–L ) (Subject 5 in Table 3 ) ( A , B , G and H ) Haematoxylin and eosin stain showing vacuolation. The 3F4 immunohistochemistry stain ( C , D , I and J) shows finely granular and coarse Pri sca deposition throughout the cerebral cortex in the case without white matter staining ( C ) and diffuse granular staining throughout the cortex in the case with white matter PrP Sc ( I ). No PrP Sc is found in the deep cerebral white matter ( D ). PrP Sc is seen in layer 6 of the cerebral cortex (CC, upper 20% of J ) and in the subcortical (lower 80% of J ) and deep white matter (not shown). Anti-GFAP antibody immunohistoc hemistry stain ( E , F , K and L ) shows more intense reactive astrocytic gliosis in the white matter ( F and L ) than in the cerebral cortex ( E and K : panels show only layers 1 to 3). Scale bar: H = 100 µm, and applies to all panels. WM = white matter; CC = cerebral cortex grey matter.
    Figure Legend Snippet: Histopathology findings. Examples of the main neuropathological changes found in sporadic CJD cases without white matter PrP Sc ( A–F , Subject 18, see Table 3 ) and with mild white matter PrP Sc ( G–L ) (Subject 5 in Table 3 ) ( A , B , G and H ) Haematoxylin and eosin stain showing vacuolation. The 3F4 immunohistochemistry stain ( C , D , I and J) shows finely granular and coarse Pri sca deposition throughout the cerebral cortex in the case without white matter staining ( C ) and diffuse granular staining throughout the cortex in the case with white matter PrP Sc ( I ). No PrP Sc is found in the deep cerebral white matter ( D ). PrP Sc is seen in layer 6 of the cerebral cortex (CC, upper 20% of J ) and in the subcortical (lower 80% of J ) and deep white matter (not shown). Anti-GFAP antibody immunohistoc hemistry stain ( E , F , K and L ) shows more intense reactive astrocytic gliosis in the white matter ( F and L ) than in the cerebral cortex ( E and K : panels show only layers 1 to 3). Scale bar: H = 100 µm, and applies to all panels. WM = white matter; CC = cerebral cortex grey matter.

    Techniques Used: Histopathology, H&E Stain, Immunohistochemistry, Staining

    26) Product Images from "Distribution and localization Patterns of Estrogen Receptor-? and Insulin-Like Growth Factor-1 Receptors in Neurons and Glial Cells of the Female Rat Substantia Nigra"

    Article Title: Distribution and localization Patterns of Estrogen Receptor-? and Insulin-Like Growth Factor-1 Receptors in Neurons and Glial Cells of the Female Rat Substantia Nigra

    Journal: The Journal of comparative neurology

    doi: 10.1002/cne.21358

    Proposed model for the neuroprotective interactions between E2 and IGF-1 on SNpc DA neurons. A) E2 and IGF-1 can directly protect the subset of SNpc DA neurons that express ERβ (green labeled nucleus) or IGF-1R (red dots). B) E2 and IGF-1 can stimulate secretion of neuroprotective factors (e.g. IGF-1; yellow filled circles) in glial cells that express ERβ, resulting in indirect neuroprotection of SNpc DA neurons that are ERβ negative.
    Figure Legend Snippet: Proposed model for the neuroprotective interactions between E2 and IGF-1 on SNpc DA neurons. A) E2 and IGF-1 can directly protect the subset of SNpc DA neurons that express ERβ (green labeled nucleus) or IGF-1R (red dots). B) E2 and IGF-1 can stimulate secretion of neuroprotective factors (e.g. IGF-1; yellow filled circles) in glial cells that express ERβ, resulting in indirect neuroprotection of SNpc DA neurons that are ERβ negative.

    Techniques Used: Labeling

    Immunoflourescent photomicrographs demonstrating differences in localization of ERβ-ir between Z8P antibody (A) and Ab-1 (E) on TH-ir SNpc DA neurons. (B–D) Enhanced magnification of ERβ and TH colocalization. TH (red, B), Z8P (green, C). Overlay (D) of B and C; arrow demonstrates nuclear labeling. (F–H) Enhanced magnification of ERβ and TH colocalization. TH (red, F), anjd Ab-1 antibody (green, G). Overlay (H) of F and G; arrow demonstrates cytoplasmic labeling. Scale bar = 10 µm.
    Figure Legend Snippet: Immunoflourescent photomicrographs demonstrating differences in localization of ERβ-ir between Z8P antibody (A) and Ab-1 (E) on TH-ir SNpc DA neurons. (B–D) Enhanced magnification of ERβ and TH colocalization. TH (red, B), Z8P (green, C). Overlay (D) of B and C; arrow demonstrates nuclear labeling. (F–H) Enhanced magnification of ERβ and TH colocalization. TH (red, F), anjd Ab-1 antibody (green, G). Overlay (H) of F and G; arrow demonstrates cytoplasmic labeling. Scale bar = 10 µm.

    Techniques Used: Labeling

    Immunoflourescent photomicrographs from substantia nigra pars compacta sections. (A) TH (DAB), black dashed lines depicts SNpc. (B) TH (Texas Red), white dashed lined depicts SNpc. (C) Fluorogold retrograde staining. (D) Immunoflourescent product for IGF-1R (green). (E) Immunoflourescent product for Z8P ERβ (Green). (F) Ab-1 ERβ (Green). (A) Scale bar = 200 µm. (B–F) Scare bar = 50 µm.
    Figure Legend Snippet: Immunoflourescent photomicrographs from substantia nigra pars compacta sections. (A) TH (DAB), black dashed lines depicts SNpc. (B) TH (Texas Red), white dashed lined depicts SNpc. (C) Fluorogold retrograde staining. (D) Immunoflourescent product for IGF-1R (green). (E) Immunoflourescent product for Z8P ERβ (Green). (F) Ab-1 ERβ (Green). (A) Scale bar = 200 µm. (B–F) Scare bar = 50 µm.

    Techniques Used: Staining

    Confocal photomicrographs from substantia nigra sections showing immunofluorescence for GFAP (red, A and D), ERβ (green, B) or IGF-1R (green, E), and their colocalization (yellow, C and F). Magnified image in (C) displays nuclear, cytoplasmic and processes immunostaining for ERβ. Scale bar = 50 µm.
    Figure Legend Snippet: Confocal photomicrographs from substantia nigra sections showing immunofluorescence for GFAP (red, A and D), ERβ (green, B) or IGF-1R (green, E), and their colocalization (yellow, C and F). Magnified image in (C) displays nuclear, cytoplasmic and processes immunostaining for ERβ. Scale bar = 50 µm.

    Techniques Used: Immunofluorescence, Immunostaining

    Low (A) and high-power (B) photomicrographs illustrating the distribution of nickel DAB ERβ-ir on TH-ir SNpc DA neurons. (B) Arrowheads point to TH-ir neurons displaying nuclear ERβ-ir, arrows demonstrate no nuclear ERβ-ir. Low (C) and high-power (D) photomicrographs illustrating the distribution of nickel DAB IGF-1R-ir in TH-ir SNpc DA neurons. (D) Arrowheads, demonstrate IGF-1R-ir in TH-ir SNpc DA neurons, arrows demonstrate no IGF-1R-ir. Scale bar = 50 µm (A and C). Scale bar = 10 µm (B and D).
    Figure Legend Snippet: Low (A) and high-power (B) photomicrographs illustrating the distribution of nickel DAB ERβ-ir on TH-ir SNpc DA neurons. (B) Arrowheads point to TH-ir neurons displaying nuclear ERβ-ir, arrows demonstrate no nuclear ERβ-ir. Low (C) and high-power (D) photomicrographs illustrating the distribution of nickel DAB IGF-1R-ir in TH-ir SNpc DA neurons. (D) Arrowheads, demonstrate IGF-1R-ir in TH-ir SNpc DA neurons, arrows demonstrate no IGF-1R-ir. Scale bar = 50 µm (A and C). Scale bar = 10 µm (B and D).

    Techniques Used:

    27) Product Images from "Tumor necrosis factor-? receptor type 1, not type 2, mediates its acute responses in the kidney"

    Article Title: Tumor necrosis factor-? receptor type 1, not type 2, mediates its acute responses in the kidney

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00426.2011

    Immunoexpression for TNF-α receptor types 1 and 2 (TNFαR1 and TNFαR2) in renal tissues. A and B : TNFαR1 immunoexpression in renal slices from WT mice. Sections were incubated with a rabbit polyclonal antibody (catalog no.
    Figure Legend Snippet: Immunoexpression for TNF-α receptor types 1 and 2 (TNFαR1 and TNFαR2) in renal tissues. A and B : TNFαR1 immunoexpression in renal slices from WT mice. Sections were incubated with a rabbit polyclonal antibody (catalog no.

    Techniques Used: Mouse Assay, Incubation

    28) Product Images from "Pharmacologic inhibition of the enzymatic effects of tissue transglutaminase reduces cardiac fibrosis and attenuates cardiomyocyte hypertrophy following pressure overload"

    Article Title: Pharmacologic inhibition of the enzymatic effects of tissue transglutaminase reduces cardiac fibrosis and attenuates cardiomyocyte hypertrophy following pressure overload

    Journal: Journal of molecular and cellular cardiology

    doi: 10.1016/j.yjmcc.2018.02.016

    tTG activation in the pressure-overloaded myocardium is markedly attenuated following daily injection of the specific small molecule inhibitor ERW1041E (ERW) 5-biotinamidopentylamine (5BP), a synthetic substrate of tTG, was used as a probe for tTG activity in tissues. When administered systemically, 5BP is covalently attached to extracellular matrix proteins in tissues in which tTG is enzymatically active. 5BP injections were performed in mice undergoing TAC protocols and sham controls. 5BP was visualized in frozen sections using a biotin-avidin-peroxidase method and was developed with DAB+nickel (black). Representative images from sham (A, D, G, J), TAC (B, E, H, K) and ERW-treated TAC animals (C, F, I, L) are shown. Sham hearts showed negligible tTG activity (A, D, G, J). After 14 days of TAC, intense staining, reflecting tTG activation was noted in areas of perivascular fibrosis (B, E, arrows), in the interstitium (H, K, arrows) and in cardiomyocytes. Treatment with ERW markedly attenuated tTG activity (C, F, I. L). M: Semiquantitive analysis showed marked reduction in tTG activity in ERW-treated animals after 14 days of TAC (***p
    Figure Legend Snippet: tTG activation in the pressure-overloaded myocardium is markedly attenuated following daily injection of the specific small molecule inhibitor ERW1041E (ERW) 5-biotinamidopentylamine (5BP), a synthetic substrate of tTG, was used as a probe for tTG activity in tissues. When administered systemically, 5BP is covalently attached to extracellular matrix proteins in tissues in which tTG is enzymatically active. 5BP injections were performed in mice undergoing TAC protocols and sham controls. 5BP was visualized in frozen sections using a biotin-avidin-peroxidase method and was developed with DAB+nickel (black). Representative images from sham (A, D, G, J), TAC (B, E, H, K) and ERW-treated TAC animals (C, F, I, L) are shown. Sham hearts showed negligible tTG activity (A, D, G, J). After 14 days of TAC, intense staining, reflecting tTG activation was noted in areas of perivascular fibrosis (B, E, arrows), in the interstitium (H, K, arrows) and in cardiomyocytes. Treatment with ERW markedly attenuated tTG activity (C, F, I. L). M: Semiquantitive analysis showed marked reduction in tTG activity in ERW-treated animals after 14 days of TAC (***p

    Techniques Used: Activation Assay, Injection, Activity Assay, Mouse Assay, Avidin-Biotin Assay, Staining

    29) Product Images from "Guard Cell- and Phloem Idioblast-Specific Expression of Thioglucoside Glucohydrolase 1 (Myrosinase) in Arabidopsis 1"

    Article Title: Guard Cell- and Phloem Idioblast-Specific Expression of Thioglucoside Glucohydrolase 1 (Myrosinase) in Arabidopsis 1

    Journal: Plant Physiology

    doi: 10.1104/pp.010925

    Immunocytochemical localization of myrosinase in transverse and longitual sections of Arabidopsis flower stalks using the anti-myrosinase antibodies K089 and 3D7. A, B, and D, Transverse sections. C, Longitudinal section. A and B, Sections immunolabeled with the 3D7 antibody. C and D, Sections immunolabeled with the K089 antibody. Arrows indicate myrosinase-containing MGCs and MPCs. Bars = 20 μm.
    Figure Legend Snippet: Immunocytochemical localization of myrosinase in transverse and longitual sections of Arabidopsis flower stalks using the anti-myrosinase antibodies K089 and 3D7. A, B, and D, Transverse sections. C, Longitudinal section. A and B, Sections immunolabeled with the 3D7 antibody. C and D, Sections immunolabeled with the K089 antibody. Arrows indicate myrosinase-containing MGCs and MPCs. Bars = 20 μm.

    Techniques Used: Immunolabeling

    30) Product Images from "A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71"

    Article Title: A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71

    Journal: Journal of Virology

    doi: 10.1128/JVI.00183-19

    Expression of mSCARB2/hSCARB2 in Scarb2-SCARB2 BAC Tg mice. (A) Schematic diagram showing the construction of the recombinant BAC. The RP23-228N3 clone is shown at the top. The genomic sequence spanning from exon 3 to exon 12 was replaced with the corresponding part of human SCARB2 cDNA that carries a FLAG tag-coding sequence. The resulting clone carrying the humanized Scarb2 transgene is displayed at the bottom. PCR genotyping of the Tg mice is also shown. Genomic DNAs isolated from Tg3 and non-Tg mice were subjected to PCR genotyping as described in Materials and Methods. The amplified product for the Tg sample is 1,511 bp long. NTC, no-template control. (B) Tissue distribution of the mSCARB2/hSCARB2 chimeric protein in Tg mice. (Top and middle) Tg and non-Tg mice were sacrificed, and cerebrum, cerebellum, spinal cord, heart, liver, spleen, lung, stomach, duodenum, jejunum, ileum, colon, kidney, and skeletal muscle were harvested for Western blotting with anti-hSCARB2 (top) and anti-FLAG (middle) antibodies. It should be noted that the anti-hSCARB2 antibody is known to cross-react with the murine endogenous protein. (Bottom) The immunoblot membrane shown in the top panel was stripped and reprobed with anti-SCARB2 antibody, which can react with both murine and human proteins. A representative result out of three experiments is shown. (C) IHC analysis of expression of the chimeric protein in Tg mice. Cerebral cortex, hippocampus, hypothalamus, medulla, thoracic spinal cord, stomach, duodenum, jejunum, ileum, colon, liver, spleen, skeletal muscle, thymus, and mesenteric lymph nodes were collected from the Tg mice; paraffinized; sectioned for immunohistochemical staining with the anti-hSCARB2 antibody; and counterstained with hematoxylin. Original magnification, ×200. A representative result out of three experiments is shown.
    Figure Legend Snippet: Expression of mSCARB2/hSCARB2 in Scarb2-SCARB2 BAC Tg mice. (A) Schematic diagram showing the construction of the recombinant BAC. The RP23-228N3 clone is shown at the top. The genomic sequence spanning from exon 3 to exon 12 was replaced with the corresponding part of human SCARB2 cDNA that carries a FLAG tag-coding sequence. The resulting clone carrying the humanized Scarb2 transgene is displayed at the bottom. PCR genotyping of the Tg mice is also shown. Genomic DNAs isolated from Tg3 and non-Tg mice were subjected to PCR genotyping as described in Materials and Methods. The amplified product for the Tg sample is 1,511 bp long. NTC, no-template control. (B) Tissue distribution of the mSCARB2/hSCARB2 chimeric protein in Tg mice. (Top and middle) Tg and non-Tg mice were sacrificed, and cerebrum, cerebellum, spinal cord, heart, liver, spleen, lung, stomach, duodenum, jejunum, ileum, colon, kidney, and skeletal muscle were harvested for Western blotting with anti-hSCARB2 (top) and anti-FLAG (middle) antibodies. It should be noted that the anti-hSCARB2 antibody is known to cross-react with the murine endogenous protein. (Bottom) The immunoblot membrane shown in the top panel was stripped and reprobed with anti-SCARB2 antibody, which can react with both murine and human proteins. A representative result out of three experiments is shown. (C) IHC analysis of expression of the chimeric protein in Tg mice. Cerebral cortex, hippocampus, hypothalamus, medulla, thoracic spinal cord, stomach, duodenum, jejunum, ileum, colon, liver, spleen, skeletal muscle, thymus, and mesenteric lymph nodes were collected from the Tg mice; paraffinized; sectioned for immunohistochemical staining with the anti-hSCARB2 antibody; and counterstained with hematoxylin. Original magnification, ×200. A representative result out of three experiments is shown.

    Techniques Used: Expressing, BAC Assay, Mouse Assay, Recombinant, Sequencing, FLAG-tag, Polymerase Chain Reaction, Isolation, Amplification, Western Blot, Immunohistochemistry, Staining

    31) Product Images from "Novel Chlamydia muridarum T cell Antigens Induce Protective Immunity against Lung and Genital Tract Infection in Murine Models"

    Article Title: Novel Chlamydia muridarum T cell Antigens Induce Protective Immunity against Lung and Genital Tract Infection in Murine Models

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi:

    Chlamydia protein uptake by DCs detected by immunohistochemistry Chlamydia protein uptake in DCs transfected with PmpG-1 25–500 , Rplf, PmpE/F-2 25–575 , MOMP or without protein. The cationic liposome DOTAP was used to deliver Chlamydia protein into the DCs. The presence of Chlamydia protein after transfection was visualized with a protein corresponding polyclonal mouse antibody followed by a biotinylated horse anti-mouse IgG and a DAB substrate.
    Figure Legend Snippet: Chlamydia protein uptake by DCs detected by immunohistochemistry Chlamydia protein uptake in DCs transfected with PmpG-1 25–500 , Rplf, PmpE/F-2 25–575 , MOMP or without protein. The cationic liposome DOTAP was used to deliver Chlamydia protein into the DCs. The presence of Chlamydia protein after transfection was visualized with a protein corresponding polyclonal mouse antibody followed by a biotinylated horse anti-mouse IgG and a DAB substrate.

    Techniques Used: Immunohistochemistry, Transfection

    32) Product Images from "Upregulation of GRAIL Is Associated with Impaired CD4 T-Cell Proliferation in Sepsis"

    Article Title: Upregulation of GRAIL Is Associated with Impaired CD4 T-Cell Proliferation in Sepsis

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1302160

    Upregulation of GRAIL in CD4 T-cells following sepsis (A) After 24 h of CLP, spleen tissues were harvested from shams and septic mice and fixed in 10% formalin and embedded in paraffin. Tissue blocks were sectioned at a thickness of 5 μm and incubated with rabbit anti-mouse GRAIL Ab, followed by incubation with biotinylated anti-rabbit IgG. Staining was developed by Vectastain ABC reagent and a diaminobenzidine kit. Representative immunostaing images at original magnification of ×100 and ×200 ( inset ) are shown. (B) Cells were isolated from spleens of sham and CLP animals; a total of 2 ×10 6 cells were used to extract mRNA and subsequent realtime PCR analysis using GRAIL primer. The expression of β-actin serves as an internal control. The data is expressed as fold induction in comparison to the shams. (C) Splenocytes (1 × 10 6 cells) from the shams and septic mice were stained for surface CD4 antigen and intracellular GRAIL using PerCP/Cy5.5-CD4 Ab, and rabbit anti-GRAIL primary Ab, respectively. A FITC-labeled anti-rabbit secondary Ab was used subsequently and then events were acquired in flow cytometer. (D) Bar diagram representing the mean fluoresce intensities of sham and septic samples are shown. Data are expressed as means ± SE (n=5 mice/group). * P
    Figure Legend Snippet: Upregulation of GRAIL in CD4 T-cells following sepsis (A) After 24 h of CLP, spleen tissues were harvested from shams and septic mice and fixed in 10% formalin and embedded in paraffin. Tissue blocks were sectioned at a thickness of 5 μm and incubated with rabbit anti-mouse GRAIL Ab, followed by incubation with biotinylated anti-rabbit IgG. Staining was developed by Vectastain ABC reagent and a diaminobenzidine kit. Representative immunostaing images at original magnification of ×100 and ×200 ( inset ) are shown. (B) Cells were isolated from spleens of sham and CLP animals; a total of 2 ×10 6 cells were used to extract mRNA and subsequent realtime PCR analysis using GRAIL primer. The expression of β-actin serves as an internal control. The data is expressed as fold induction in comparison to the shams. (C) Splenocytes (1 × 10 6 cells) from the shams and septic mice were stained for surface CD4 antigen and intracellular GRAIL using PerCP/Cy5.5-CD4 Ab, and rabbit anti-GRAIL primary Ab, respectively. A FITC-labeled anti-rabbit secondary Ab was used subsequently and then events were acquired in flow cytometer. (D) Bar diagram representing the mean fluoresce intensities of sham and septic samples are shown. Data are expressed as means ± SE (n=5 mice/group). * P

    Techniques Used: Mouse Assay, Incubation, Staining, Isolation, Polymerase Chain Reaction, Expressing, Labeling, Flow Cytometry, Cytometry

    33) Product Images from "Distribution and localization Patterns of Estrogen Receptor-? and Insulin-Like Growth Factor-1 Receptors in Neurons and Glial Cells of the Female Rat Substantia Nigra"

    Article Title: Distribution and localization Patterns of Estrogen Receptor-? and Insulin-Like Growth Factor-1 Receptors in Neurons and Glial Cells of the Female Rat Substantia Nigra

    Journal: The Journal of comparative neurology

    doi: 10.1002/cne.21358

    Low (A) and high-power (B) photomicrographs illustrating the distribution of nickel DAB ERβ-ir on TH-ir SNpc DA neurons. (B) Arrowheads point to TH-ir neurons displaying nuclear ERβ-ir, arrows demonstrate no nuclear ERβ-ir. Low (C) and high-power (D) photomicrographs illustrating the distribution of nickel DAB IGF-1R-ir in TH-ir SNpc DA neurons. (D) Arrowheads, demonstrate IGF-1R-ir in TH-ir SNpc DA neurons, arrows demonstrate no IGF-1R-ir. Scale bar = 50 µm (A and C). Scale bar = 10 µm (B and D).
    Figure Legend Snippet: Low (A) and high-power (B) photomicrographs illustrating the distribution of nickel DAB ERβ-ir on TH-ir SNpc DA neurons. (B) Arrowheads point to TH-ir neurons displaying nuclear ERβ-ir, arrows demonstrate no nuclear ERβ-ir. Low (C) and high-power (D) photomicrographs illustrating the distribution of nickel DAB IGF-1R-ir in TH-ir SNpc DA neurons. (D) Arrowheads, demonstrate IGF-1R-ir in TH-ir SNpc DA neurons, arrows demonstrate no IGF-1R-ir. Scale bar = 50 µm (A and C). Scale bar = 10 µm (B and D).

    Techniques Used:

    34) Product Images from "Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis"

    Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/bcr3053

    Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.
    Figure Legend Snippet: Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.

    Techniques Used: Expressing, Staining, Immunohistochemistry, Microscopy

    35) Product Images from "Primate-specific Melanoma Antigen-A11 Regulates Isoform-specific Human Progesterone Receptor-B Transactivation *"

    Article Title: Primate-specific Melanoma Antigen-A11 Regulates Isoform-specific Human Progesterone Receptor-B Transactivation *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M112.372797

    FKBP5, MAGE-11, PR, and p300 mRNA levels in normal human endometrium through the menstrual cycle. Quantitative real-time RT-PCR of FKBP5 ( A ), MAGE-11 ( B ), PR ( C ), and p300 ( D ) mRNA was performed using human endometrium tissue from normal cycling women
    Figure Legend Snippet: FKBP5, MAGE-11, PR, and p300 mRNA levels in normal human endometrium through the menstrual cycle. Quantitative real-time RT-PCR of FKBP5 ( A ), MAGE-11 ( B ), PR ( C ), and p300 ( D ) mRNA was performed using human endometrium tissue from normal cycling women

    Techniques Used: Quantitative RT-PCR

    Schematic diagram of PR-B, MAGE-11, FKBP5, and p300 transcriptional complex. MAGE-11 F-box and lysine ubiquitinylation sites interact with the PR-B NH 2 -terminal 110 LL XX VL XX LL 119 motif region absent in PR-A. MAGE-11 residues 252–276 and residues
    Figure Legend Snippet: Schematic diagram of PR-B, MAGE-11, FKBP5, and p300 transcriptional complex. MAGE-11 F-box and lysine ubiquitinylation sites interact with the PR-B NH 2 -terminal 110 LL XX VL XX LL 119 motif region absent in PR-A. MAGE-11 residues 252–276 and residues

    Techniques Used:

    Interaction domains in PR-B and MAGE-11. A , pCMV-FLAG empty vector (−) (5 μg) and 5 μg of pCMV-FLAG-PR-B-(1–164) wild type ( WT ) and L110A,L111A, V114A,L115A, and L118A,L119A mutants were expressed in COS cells with 4 μg
    Figure Legend Snippet: Interaction domains in PR-B and MAGE-11. A , pCMV-FLAG empty vector (−) (5 μg) and 5 μg of pCMV-FLAG-PR-B-(1–164) wild type ( WT ) and L110A,L111A, V114A,L115A, and L118A,L119A mutants were expressed in COS cells with 4 μg

    Techniques Used: Plasmid Preparation

    Activation of unique PR-B NH 2 -terminal region by MAGE-11 and p300. A and B , GAL-PR-B-(1–164) (50 ng) was expressed in HeLa cells with 0.1 μg of 5XGAL4Luc3 and 10 or 150 ng of pSG5 empty vector, 10–150 ng of pSG5-MAGE ( A ), and 10–150
    Figure Legend Snippet: Activation of unique PR-B NH 2 -terminal region by MAGE-11 and p300. A and B , GAL-PR-B-(1–164) (50 ng) was expressed in HeLa cells with 0.1 μg of 5XGAL4Luc3 and 10 or 150 ng of pSG5 empty vector, 10–150 ng of pSG5-MAGE ( A ), and 10–150

    Techniques Used: Activation Assay, Plasmid Preparation

    MAGE-11 stabilizes a complex with PR-B and FKBP5. In A–D , pCMV-FLAG empty vector (3 or 4 μg) (−) and 3 and 4 μg of FLAG-FKBP5 were expressed in COS cells with the following DNAs. A , 2 μg of pSG5-MAGE with and without
    Figure Legend Snippet: MAGE-11 stabilizes a complex with PR-B and FKBP5. In A–D , pCMV-FLAG empty vector (3 or 4 μg) (−) and 3 and 4 μg of FLAG-FKBP5 were expressed in COS cells with the following DNAs. A , 2 μg of pSG5-MAGE with and without

    Techniques Used: Plasmid Preparation

    Requirement for MAGE-11 in PR-B up-regulation of FKBP5 but not RASD1 . A , top , IKPRB cells were treated in charcoal-stripped serum-containing medium with 20 n m R5020 for 0, 8, and 24 h. Cell extracts (40 μg of protein for FKBP5 and β-actin,
    Figure Legend Snippet: Requirement for MAGE-11 in PR-B up-regulation of FKBP5 but not RASD1 . A , top , IKPRB cells were treated in charcoal-stripped serum-containing medium with 20 n m R5020 for 0, 8, and 24 h. Cell extracts (40 μg of protein for FKBP5 and β-actin,

    Techniques Used:

    MAGE-11 effects independent of PR-B terminal phosphorylation. A , schematic diagram of human PR-B, which contains the NH 2 -terminal 110 LL XX VL XX LL 119 motif, DNA-binding domain ( DBD ), ligand-binding domain ( LBD ), and PR-B upstream segment ( BUS ) residues 1–168
    Figure Legend Snippet: MAGE-11 effects independent of PR-B terminal phosphorylation. A , schematic diagram of human PR-B, which contains the NH 2 -terminal 110 LL XX VL XX LL 119 motif, DNA-binding domain ( DBD ), ligand-binding domain ( LBD ), and PR-B upstream segment ( BUS ) residues 1–168

    Techniques Used: Binding Assay, Ligand Binding Assay

    Immunostaining of MAGE-11, FKBP5, p300, and PR in normal human endometrium through the menstrual cycle. Immunostaining was performed as described under “Experimental Procedures” on serial sections of human endometrium from different menstrual
    Figure Legend Snippet: Immunostaining of MAGE-11, FKBP5, p300, and PR in normal human endometrium through the menstrual cycle. Immunostaining was performed as described under “Experimental Procedures” on serial sections of human endometrium from different menstrual

    Techniques Used: Immunostaining

    Dependence on PR-B NH 2 -terminal 110 LL XX VL XX LL 119 motif for down-regulation and transcriptional activation by MAGE-11. A , human PR-B NH 2 -terminal 164 amino acid residues contain activation function 3 ( AF-3 ) and an 110 LL XX VL XX LL 119 motif not present in
    Figure Legend Snippet: Dependence on PR-B NH 2 -terminal 110 LL XX VL XX LL 119 motif for down-regulation and transcriptional activation by MAGE-11. A , human PR-B NH 2 -terminal 164 amino acid residues contain activation function 3 ( AF-3 ) and an 110 LL XX VL XX LL 119 motif not present in

    Techniques Used: Activation Assay

    Interaction between PR-B and MAGE-11 up-regulates FKBP5 progesterone response region. A , p5M-PR-B and -A (0.5 μg) were expressed with 1 μg of pCMV-FLAG empty vector or 1 μg of pCMV-FLAG-MAGE in COS cells. The day after transfection,
    Figure Legend Snippet: Interaction between PR-B and MAGE-11 up-regulates FKBP5 progesterone response region. A , p5M-PR-B and -A (0.5 μg) were expressed with 1 μg of pCMV-FLAG empty vector or 1 μg of pCMV-FLAG-MAGE in COS cells. The day after transfection,

    Techniques Used: Plasmid Preparation, Transfection

    36) Product Images from "Pharmacological rescue of the brain cortex phenotype of Tbx1 mouse mutants: significance for 22q11.2 deletion syndrome"

    Article Title: Pharmacological rescue of the brain cortex phenotype of Tbx1 mouse mutants: significance for 22q11.2 deletion syndrome

    Journal: bioRxiv

    doi: 10.1101/2021.02.04.429794

    Vitamin B12 treatment restores normal expression of cortical markers in Tbx1 heterozygous embryos. Panels show coronal sections of embryonic brains at E13.5. (A, a) Immunostaining for TBR1 (A–A′′’) revealed the precocious presence of terminally differentiated neurons in the SVZ of Tbx1 lacZ /+ embryos (A’). White brackets define the regions containing TBR1-positive cells in A, A’. Quantitative analysis confirmed the altered distribution and frequency of mature cortical neurons in the ML-cortex, of Tbx1 lacZ /+ embryos, including the SVZ and CP (a), which was rescued by B12 treatment (A”’, a). (B, b) Immunostaining for phospho-histone 3 (PH3) revealed increased mitotic activity in the proliferative zones (VZ, SVZ) of the ML-cortex of vehicle-treated Tbx1 lacZ /+ embryos (B’, b, b’) compared to vehicle-treated WT embryos (B, b), which was rescued by B12 treatment (B”’, b, b’). (C, c) Immunostaining for KI67. White asterisks identify KI67-negative regions. Quantitative analysis (c) revealed the reduced thickness of the KI67-expressing region (yellow brackets) relative to the total ventricular-to-pial thickness (white brackets) in vehicle-treated Tbx1 lacZ /+ embryos (C’, c) compared to vehicle-treated WT embryos (C), which was rescued by B12 treatment (C”’, c). The cartoon (c’) indicates the position of the ML-cortex in coronal sections, where all measurements were made. Scale bars, 100 μm. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, error bars indicate ±SEM, n = 5–6 per genotype. Abbreviations: VZ, ventricular zone; SVZ, subventricular zone; CP, cortical plate; ML-Cx, medio-lateral cortex; D, dorsal; M, medial; ML, medio-lateral; L, lateral; OD, optical density.
    Figure Legend Snippet: Vitamin B12 treatment restores normal expression of cortical markers in Tbx1 heterozygous embryos. Panels show coronal sections of embryonic brains at E13.5. (A, a) Immunostaining for TBR1 (A–A′′’) revealed the precocious presence of terminally differentiated neurons in the SVZ of Tbx1 lacZ /+ embryos (A’). White brackets define the regions containing TBR1-positive cells in A, A’. Quantitative analysis confirmed the altered distribution and frequency of mature cortical neurons in the ML-cortex, of Tbx1 lacZ /+ embryos, including the SVZ and CP (a), which was rescued by B12 treatment (A”’, a). (B, b) Immunostaining for phospho-histone 3 (PH3) revealed increased mitotic activity in the proliferative zones (VZ, SVZ) of the ML-cortex of vehicle-treated Tbx1 lacZ /+ embryos (B’, b, b’) compared to vehicle-treated WT embryos (B, b), which was rescued by B12 treatment (B”’, b, b’). (C, c) Immunostaining for KI67. White asterisks identify KI67-negative regions. Quantitative analysis (c) revealed the reduced thickness of the KI67-expressing region (yellow brackets) relative to the total ventricular-to-pial thickness (white brackets) in vehicle-treated Tbx1 lacZ /+ embryos (C’, c) compared to vehicle-treated WT embryos (C), which was rescued by B12 treatment (C”’, c). The cartoon (c’) indicates the position of the ML-cortex in coronal sections, where all measurements were made. Scale bars, 100 μm. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, error bars indicate ±SEM, n = 5–6 per genotype. Abbreviations: VZ, ventricular zone; SVZ, subventricular zone; CP, cortical plate; ML-Cx, medio-lateral cortex; D, dorsal; M, medial; ML, medio-lateral; L, lateral; OD, optical density.

    Techniques Used: Expressing, Immunostaining, Activity Assay

    37) Product Images from "Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis"

    Article Title: Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/bcr3053

    Down-regulation of IRF5 protein expression by siRNAs alters sensitivity to DNA damage . A . MCF-12A cells were incubated with transfection reagent alone (mock-transfected), control Lamin A/C siRNAs or 5 nM IRF5 siRNAs once (IRF5 siRNA1) or twice (IRF5 siRNA2), as described in the Materials and methods. Western blot analysis shows > 70% reduction of endogenous IRF5 proteins after normalization to β-actin levels. B . Cells were exposed to 5 Gy IR or the same dose plus IFN-γ (IR/γ) for 24 h. Percent of Annexin V-FITC stained positive cells is shown in the upper and lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. C . Same as in B, except cells were exposed to 1 μM Dox or Dox and IFN-γ for five hours. Percent of Annexin V-FITC stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between cells transfected with Lamin A/C siRNAs (12Asicon) and IRF5 siRNAs (12AsiIRF5) after each treatment; ** denotes P
    Figure Legend Snippet: Down-regulation of IRF5 protein expression by siRNAs alters sensitivity to DNA damage . A . MCF-12A cells were incubated with transfection reagent alone (mock-transfected), control Lamin A/C siRNAs or 5 nM IRF5 siRNAs once (IRF5 siRNA1) or twice (IRF5 siRNA2), as described in the Materials and methods. Western blot analysis shows > 70% reduction of endogenous IRF5 proteins after normalization to β-actin levels. B . Cells were exposed to 5 Gy IR or the same dose plus IFN-γ (IR/γ) for 24 h. Percent of Annexin V-FITC stained positive cells is shown in the upper and lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. C . Same as in B, except cells were exposed to 1 μM Dox or Dox and IFN-γ for five hours. Percent of Annexin V-FITC stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between cells transfected with Lamin A/C siRNAs (12Asicon) and IRF5 siRNAs (12AsiIRF5) after each treatment; ** denotes P

    Techniques Used: Expressing, Incubation, Transfection, Western Blot, Staining

    IRF5 reduces CXCR4 cell surface expression and SDF-1/CXCL12-dependent chemotaxis of MDA-MB-231 cells . A . CXCR4 expression (grey line) in unstimulated cells, shown superimposed on the isotype control (grey shaded area), and CXCR4 expression (black line) after stimulation, was measured by flow cytometry. MDA-MB-231 cells (pBabe and pBIRF5) were treated with the CXCR4 ligand SDF-1 for six hours and CXCR4 expression measured. IRF5 expressing cells show no significant expression of CXCR4. M1, Marker 1. Representative histogram plots from three independent experiments performed in duplicate are shown. B . Cells overexpressing IRF5 are incapable of SDF-1-induced migration when compared to empty vector (EV pBabe) control cells. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference in number of cells migrated between pBabe and pBIRF5 cells; * denotes P
    Figure Legend Snippet: IRF5 reduces CXCR4 cell surface expression and SDF-1/CXCL12-dependent chemotaxis of MDA-MB-231 cells . A . CXCR4 expression (grey line) in unstimulated cells, shown superimposed on the isotype control (grey shaded area), and CXCR4 expression (black line) after stimulation, was measured by flow cytometry. MDA-MB-231 cells (pBabe and pBIRF5) were treated with the CXCR4 ligand SDF-1 for six hours and CXCR4 expression measured. IRF5 expressing cells show no significant expression of CXCR4. M1, Marker 1. Representative histogram plots from three independent experiments performed in duplicate are shown. B . Cells overexpressing IRF5 are incapable of SDF-1-induced migration when compared to empty vector (EV pBabe) control cells. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference in number of cells migrated between pBabe and pBIRF5 cells; * denotes P

    Techniques Used: Expressing, Chemotaxis Assay, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Marker, Migration, Plasmid Preparation

    Summary of IRF expression in breast tissue specimens . A . Percent of samples with positive staining is shown on y-axis, tissue and disease type are shown on x-axis. Number of samples positive for IRF1 or IRF5 is shown over total number of patient samples examined in each group. Statistical significance was determined by comparing the number of positive-stained samples in each disease type to positive-stained samples in normal tissues; * denotes P
    Figure Legend Snippet: Summary of IRF expression in breast tissue specimens . A . Percent of samples with positive staining is shown on y-axis, tissue and disease type are shown on x-axis. Number of samples positive for IRF1 or IRF5 is shown over total number of patient samples examined in each group. Statistical significance was determined by comparing the number of positive-stained samples in each disease type to positive-stained samples in normal tissues; * denotes P

    Techniques Used: Expressing, Staining

    Overexpression of IRF5 in MCF-7 and MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition . A . Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of β-actin are shown as loading controls. B . Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C . Cell survival was measured in MCF-7 and MDA-MB-231 pBabe cell lines by colony formation assay before and after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 and 10 Gy γ-IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P
    Figure Legend Snippet: Overexpression of IRF5 in MCF-7 and MDA-MB-231 cells sensitizes them to DNA damage-induced growth inhibition . A . Endogenous IRF expression was analyzed by Western blot in transformed mammary epithelial cell lines. Levels of β-actin are shown as loading controls. B . Western blot analysis of stable cell lines generated to overexpress retroviral pBIRF5. C . Cell survival was measured in MCF-7 and MDA-MB-231 pBabe cell lines by colony formation assay before and after treatment. Cells were treated with 0.1 or 1 μM Doxorubicin (Dox) or 2, 5 and 10 Gy γ-IR. The number of colonies is plotted on the y-axis as percent of control; 100% represents the number of colonies in empty pBabe control lines. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between colonies in pBabe versus pBIRF5 cell lines after each treatment; * denotes P

    Techniques Used: Over Expression, Multiple Displacement Amplification, Inhibition, Expressing, Western Blot, Transformation Assay, Stable Transfection, Generated, Colony Assay

    Overexpression of IRF5 in MDA-MB-231 cells sensitizes them to IR-induced apoptosis . A . MDA-MB-231 cells were exposed to 5 Gy IR or the same dose plus IFN-γ (IR/γ) for 24 h. Percent of cells undergoing apoptosis was measured by FACS analysis of Annexin V-FITC (x-axis) and PI (y-axis) double-staining. Percent of Annexin V-FITC stained positive cells is shown in the upper and lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. B . Same as in (A), except cells were treated with 1 μM Dox or the same dose plus IFN-γ (Dox/γ) for five hours. Percent of Annexin V-FITC-stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between pBabe and pBIRF5 cells lines after each treatment; ** denotes P
    Figure Legend Snippet: Overexpression of IRF5 in MDA-MB-231 cells sensitizes them to IR-induced apoptosis . A . MDA-MB-231 cells were exposed to 5 Gy IR or the same dose plus IFN-γ (IR/γ) for 24 h. Percent of cells undergoing apoptosis was measured by FACS analysis of Annexin V-FITC (x-axis) and PI (y-axis) double-staining. Percent of Annexin V-FITC stained positive cells is shown in the upper and lower right-hand quadrants. Representative histogram plots from three independent experiments performed in duplicate are shown. B . Same as in (A), except cells were treated with 1 μM Dox or the same dose plus IFN-γ (Dox/γ) for five hours. Percent of Annexin V-FITC-stained positive cells compared to control is plotted on y-axis. Data are expressed as mean ± SD of three independent experiments performed in duplicate. Statistical significance was determined by comparing the difference between pBabe and pBIRF5 cells lines after each treatment; ** denotes P

    Techniques Used: Over Expression, Multiple Displacement Amplification, FACS, Double Staining, Staining

    IRF5 inhibits in vivo tumor formation and in vitro metastasis/invasion . A . MCF-7/pBIRF5 (MCF7-IRF5) and MCF-7/pBabe (MCF7-EV) control cells were inoculated into NCr nu/nu mice. The number of mice with tumors over the total number of mice is shown. B . Same as in (A), except 3 × 10 6 MDA-MB-231 control cells (231-EV) or MDA-MB-231 IRF5 overexpressing cells (231-IRF5) were inoculated into NCr nu/nu mice and monitored over seven weeks. C . Growth of MCF-7 and MDA-MB-231 cells were examined by 3-D culture. An equal number of cells were plated and pictures taken 10 days later at 10 × magnification.
    Figure Legend Snippet: IRF5 inhibits in vivo tumor formation and in vitro metastasis/invasion . A . MCF-7/pBIRF5 (MCF7-IRF5) and MCF-7/pBabe (MCF7-EV) control cells were inoculated into NCr nu/nu mice. The number of mice with tumors over the total number of mice is shown. B . Same as in (A), except 3 × 10 6 MDA-MB-231 control cells (231-EV) or MDA-MB-231 IRF5 overexpressing cells (231-IRF5) were inoculated into NCr nu/nu mice and monitored over seven weeks. C . Growth of MCF-7 and MDA-MB-231 cells were examined by 3-D culture. An equal number of cells were plated and pictures taken 10 days later at 10 × magnification.

    Techniques Used: In Vivo, In Vitro, Mouse Assay, Multiple Displacement Amplification

    Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.
    Figure Legend Snippet: Dysregulated IRF expression in patients with ductal carcinoma . A . Normal and ADH breast tissue specimens were stained by IF or IHC. Antibodies recognizing IRF5 (FITC), IRF1 (Cy3) and DAPI for the nucleus were used for IF. For IHC, tissues were stained for IRF1 with DAB (brownish-red), IRF5 with BAP (blue), and nucleus with Fast Red mounting buffer. B . Same as in (A), except tissue samples from patients with ADH were stained by IF with IRF5 (FITC) and CK14 (Cy3) in order to confirm expression of IRF5 in myoepithelial cells. C . Same as in (A), except tissues from patients with DCIS and IDC were examined. Representative pictures of low grade and high grade DCIS are shown illustrating distinct differences between IRF1 and IRF5 expression. Images were taken on a Zeiss Axiovert Apotome microscope at 20 × or 40 × magnification. Scale bars are 50 μm.

    Techniques Used: Expressing, Staining, Immunohistochemistry, Microscopy

    38) Product Images from "PRMT1 and PRMT4 Regulate Oxidative Stress-Induced Retinal Pigment Epithelial Cell Damage in SIRT1-Dependent and SIRT1-Independent Manners"

    Article Title: PRMT1 and PRMT4 Regulate Oxidative Stress-Induced Retinal Pigment Epithelial Cell Damage in SIRT1-Dependent and SIRT1-Independent Manners

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2015/617919

    The knockdown of PRMT1 or PRMT4 attenuates oxidative stress-induced RPE cell damage and PRMT1 expression regulates SIRT1 expression. (a) ARPE-19 cells were transfected with scramble, PRMT1, or PRMT4 siRNA according to the reverse transfection method. After 36 h, cell extracts were subjected to Western blotting with the indicated antibodies. (b, c) ARPE-19 cells were transfected with scramble, PRMT1, or PRMT4 siRNA according to the reverse transfection method. After 24 h, the medium was changed to serum-free medium and 250 μ M H 2 O 2 was added for 24 h. (b) Cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P
    Figure Legend Snippet: The knockdown of PRMT1 or PRMT4 attenuates oxidative stress-induced RPE cell damage and PRMT1 expression regulates SIRT1 expression. (a) ARPE-19 cells were transfected with scramble, PRMT1, or PRMT4 siRNA according to the reverse transfection method. After 36 h, cell extracts were subjected to Western blotting with the indicated antibodies. (b, c) ARPE-19 cells were transfected with scramble, PRMT1, or PRMT4 siRNA according to the reverse transfection method. After 24 h, the medium was changed to serum-free medium and 250 μ M H 2 O 2 was added for 24 h. (b) Cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P

    Techniques Used: Expressing, Transfection, Western Blot, MTT Assay

    PRMT1 and PRMT4 expression is increased in the RPE layer of streptozotocin-treated rats. Eyeballs were enucleated from vehicle-treated and STZ-treated rats and cryosections were prepared. (a, b) PRMT1 (a) and PRMT4 (b) expressions were measured by immunohistochemistry analysis (C: choroid, RPE: retinal pigment epithelium, PL: photoreceptor layer, OLM: outer limiting membrane, ONL: outer nuclear layer, OPL: outer plexiform layer, INL: inner nuclear layer, IPL: inner plexiform layer, and GCL: ganglion-cell layer). Representative images were from at least three independent experiments. To quantify the DAB signaling, semiautomated analysis protocol was performed as described in Section 2 .
    Figure Legend Snippet: PRMT1 and PRMT4 expression is increased in the RPE layer of streptozotocin-treated rats. Eyeballs were enucleated from vehicle-treated and STZ-treated rats and cryosections were prepared. (a, b) PRMT1 (a) and PRMT4 (b) expressions were measured by immunohistochemistry analysis (C: choroid, RPE: retinal pigment epithelium, PL: photoreceptor layer, OLM: outer limiting membrane, ONL: outer nuclear layer, OPL: outer plexiform layer, INL: inner nuclear layer, IPL: inner plexiform layer, and GCL: ganglion-cell layer). Representative images were from at least three independent experiments. To quantify the DAB signaling, semiautomated analysis protocol was performed as described in Section 2 .

    Techniques Used: Expressing, Immunohistochemistry

    PRMT1 or PRMT4 overexpression increases RPE cell apoptosis, while PRMT1 overexpression decreases SIRT1 expression. (a, b) ARPE-19 cells were transfected with HA or HA-PRMT1 plasmid DNA. After 36 h, (a) cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P
    Figure Legend Snippet: PRMT1 or PRMT4 overexpression increases RPE cell apoptosis, while PRMT1 overexpression decreases SIRT1 expression. (a, b) ARPE-19 cells were transfected with HA or HA-PRMT1 plasmid DNA. After 36 h, (a) cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P

    Techniques Used: Over Expression, Expressing, Transfection, Plasmid Preparation, MTT Assay

    H 2 O 2 increases PRMT1 and PRMT4 expression but decreases SIRT1 expression. (a, b) ARPE-19 cells were treated with 250 μ M H 2 O 2 for 12 and 24 h. (a) Cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P
    Figure Legend Snippet: H 2 O 2 increases PRMT1 and PRMT4 expression but decreases SIRT1 expression. (a, b) ARPE-19 cells were treated with 250 μ M H 2 O 2 for 12 and 24 h. (a) Cell viability was measured by the MTT assay. The data represent the means ± SEM of three independent experiments, each performed in triplicate. ∗ P

    Techniques Used: Expressing, MTT Assay

    PRMT1 and PRMT4 regulate oxidative stress-induced RPE cell damage in SIRT1-dependent and SIRT1-independent manners.
    Figure Legend Snippet: PRMT1 and PRMT4 regulate oxidative stress-induced RPE cell damage in SIRT1-dependent and SIRT1-independent manners.

    Techniques Used:

    39) Product Images from "A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71"

    Article Title: A Novel Murine Model Expressing a Chimeric mSCARB2/hSCARB2 Receptor Is Highly Susceptible to Oral Infection with Clinical Isolates of Enterovirus 71

    Journal: Journal of Virology

    doi: 10.1128/JVI.00183-19

    Replication of EV71 in orally infected Tg mice. (A) Histopathological changes and expression of EV71 VP2 protein in infected mice. Two-week-old Tg mice were inoculated i.g. with the EV71-4643 strain and euthanized at 6 days postinfection. (Left) Cerebral cortex, hippocampus, cerebellum, thoracic spinal cord, pons, medulla, skeletal muscle (mice with hind-limb paralysis and moribund mice with complete paralysis), and hind-limb paw of Tg mice were retrieved for sample processing, IHC staining with the anti-EV71 VP2 antibody, and hematoxylin counterstaining. (Right) In parallel, sections were stained with H E. Original magnification, ×200. A representative result out of six experiments is shown. (B) Viral titers in various parts of the CNS and GI tract and in skeletal muscle of infected mice. Two-week-old Tg and non-Tg mice were infected i.g. with 1 × 10 8 PFU of the EV71-4643 strain and euthanized at the indicated times. Brains and GI tracts were collected and dissected into parts, as indicated in the figure. The anterior small intestine includes the duodenum and parts of the jejunum, while the posterior small intestine includes parts of the jejunum and ileum. Viral titers were determined using a plaque-forming assay and are expressed as log PFU per gram of tissue. ns, not significant; *, P  
    Figure Legend Snippet: Replication of EV71 in orally infected Tg mice. (A) Histopathological changes and expression of EV71 VP2 protein in infected mice. Two-week-old Tg mice were inoculated i.g. with the EV71-4643 strain and euthanized at 6 days postinfection. (Left) Cerebral cortex, hippocampus, cerebellum, thoracic spinal cord, pons, medulla, skeletal muscle (mice with hind-limb paralysis and moribund mice with complete paralysis), and hind-limb paw of Tg mice were retrieved for sample processing, IHC staining with the anti-EV71 VP2 antibody, and hematoxylin counterstaining. (Right) In parallel, sections were stained with H E. Original magnification, ×200. A representative result out of six experiments is shown. (B) Viral titers in various parts of the CNS and GI tract and in skeletal muscle of infected mice. Two-week-old Tg and non-Tg mice were infected i.g. with 1 × 10 8 PFU of the EV71-4643 strain and euthanized at the indicated times. Brains and GI tracts were collected and dissected into parts, as indicated in the figure. The anterior small intestine includes the duodenum and parts of the jejunum, while the posterior small intestine includes parts of the jejunum and ileum. Viral titers were determined using a plaque-forming assay and are expressed as log PFU per gram of tissue. ns, not significant; *, P  

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

    40) Product Images from "Activity of translation regulator eukaryotic elongation factor-2 kinase is increased in Parkinson disease brain and its inhibition reduces alpha synuclein toxicity"

    Article Title: Activity of translation regulator eukaryotic elongation factor-2 kinase is increased in Parkinson disease brain and its inhibition reduces alpha synuclein toxicity

    Journal: Acta Neuropathologica Communications

    doi: 10.1186/s40478-018-0554-9

    Immunostaining for phospho-eEF2 (p-eEF2, Thr56) and phospho-AS (p-ASyn, Ser129) in postmortem control and PD midbrain serial sections. a - b p-eEF2 (T56) and p-ASyn (S129) IHC in postmortem midbrain serial sections from one control ( a ) and two PD cases ( b : Table S1. (SN- substantia nigra; PAG- periaqueductal gray matter; scale bar, 100 μm; insets show 40× magnified view in each image)
    Figure Legend Snippet: Immunostaining for phospho-eEF2 (p-eEF2, Thr56) and phospho-AS (p-ASyn, Ser129) in postmortem control and PD midbrain serial sections. a - b p-eEF2 (T56) and p-ASyn (S129) IHC in postmortem midbrain serial sections from one control ( a ) and two PD cases ( b : Table S1. (SN- substantia nigra; PAG- periaqueductal gray matter; scale bar, 100 μm; insets show 40× magnified view in each image)

    Techniques Used: Immunostaining, Immunohistochemistry

    Immunostaining for phospho-eEF2 (p-eEF2, Thr56) and phospho-AS (p-ASyn, Ser129) in postmortem control and PD hippocampus serial sections. a - b p-eEF2 (T56) and p-ASyn (S129) IHC in postmortem hippocampus serial sections from one control ( a ) and two PD cases ( b : Table S1 (CA1 and CA2- hippocampal cornu ammonis fields 1 and 2 respectively; scale bar, 100 μm; insets show 40× magnified view in each image)
    Figure Legend Snippet: Immunostaining for phospho-eEF2 (p-eEF2, Thr56) and phospho-AS (p-ASyn, Ser129) in postmortem control and PD hippocampus serial sections. a - b p-eEF2 (T56) and p-ASyn (S129) IHC in postmortem hippocampus serial sections from one control ( a ) and two PD cases ( b : Table S1 (CA1 and CA2- hippocampal cornu ammonis fields 1 and 2 respectively; scale bar, 100 μm; insets show 40× magnified view in each image)

    Techniques Used: Immunostaining, Immunohistochemistry

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    Avidin-Biotin Assay:

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    Article Snippet: .. The sections were incubated in the primary antibody (1:500) at 4°C over 72 h, processed with biotinylated anti-mouse IgG (BA-2001, 1:400; Vector Laboratories), and developed with DAB to produce a brown reaction product. .. Omission of the GAD primary antibody resulted in the absence of specific staining.

    Article Title: Blood glutamate EAAT2-cell grabbing therapy in cerebral ischemia
    Article Snippet: .. Cells were washed thrice with PBS at RT and incubated with a biotinylated horse anti-mouse IgG antibody, rat adsorbed (dilution 1:200; Vector Laboratories, Burlingame, CA, USA), for 1 h at RT, followed by a second incubation with a 1:100 dilution of avidin–biotin (Vector Laboratories) for 30 min. Once washed, the samples were incubated with DyLight 594 streptavidin (diluted 1:500 in PBS; Vector Laboratories) for 30 min at RT, followed by a last incubation step with Hoechst stain (diluted 1:6000; Invitrogen) for 5 min. Fluorescence was analyzed with a confocal microscope (LEICA AOBS-SP5X; Leica, Wetzlar, Germany). .. Flow cytometry, tube formation assay, and growth factor release analysis were performed to verify that the transfected MSCs maintained their intrinsic phenotype properties.

    other:

    Article Title: Expression of ErbB4 in the neurons of Alzheimer's disease brain and APP/PS1 mice, a model of Alzheimer's disease
    Article Snippet: Vectashield (H-1000), biotinylated anti-rabbit IgG (BA-1000) and biotinylated anti-mouse IgG (BA-2001) were supplied by Vector Laboratories (Burlingame, CA, USA).

    Labeling:

    Article Title: Alpha-synuclein is strategically positioned for afferent modulation of midbrain dopamine neurons and is essential for cocaine preference
    Article Snippet: .. Peroxidase labeling of α-syn was achieved using a biotinylated horse-anti-mouse immunoglobulin G (IgG) (1:400 dil; Vector Labs; BA-2001) and of CD63 using a biotinylated donkey anti-rabbit IgG (1:400 dil; Jackson Laboratories; #711-065-152) in 0.1% BSA in 0.1 M TBS for 30 min, rinsed, and then placed in an avidin–biotin solution (1:200 dilution in 0.1 M TBS; Vector Laboratories) for 30 min. .. The peroxidase reaction product was visualized with 0.022% 3,3′-diaminobenzidine (Aldrich) and 0.003% H2 O2 in 0.1 M TBS for 6 min. For immunogold-silver visualization of TH immunoreactivity, tissue sections were incubated for 1 h in donkey anti-sheep colloidal gold (1 nm) IgG (1:50 dilution; EMS; #25820), fixed in 2% glutaraldehyde, and enhanced with a silver solution (IntenS-EM kit: EMS) for 7 min.

    Staining:

    Article Title: Blood glutamate EAAT2-cell grabbing therapy in cerebral ischemia
    Article Snippet: .. Cells were washed thrice with PBS at RT and incubated with a biotinylated horse anti-mouse IgG antibody, rat adsorbed (dilution 1:200; Vector Laboratories, Burlingame, CA, USA), for 1 h at RT, followed by a second incubation with a 1:100 dilution of avidin–biotin (Vector Laboratories) for 30 min. Once washed, the samples were incubated with DyLight 594 streptavidin (diluted 1:500 in PBS; Vector Laboratories) for 30 min at RT, followed by a last incubation step with Hoechst stain (diluted 1:6000; Invitrogen) for 5 min. Fluorescence was analyzed with a confocal microscope (LEICA AOBS-SP5X; Leica, Wetzlar, Germany). .. Flow cytometry, tube formation assay, and growth factor release analysis were performed to verify that the transfected MSCs maintained their intrinsic phenotype properties.

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    Vector Laboratories dab peroxidase
    CXCR4 and ACKR3 staining using IHC in breast cancer tissue. 4 μm sections from human breast cancer were stained for CXCR4 (1:40) and ACKR3 (1:100) using immunohistochemistry following no pre-treatment or EDTA antigen retrieval pre-treatment, respectively. Briefly, protocol from the VECTASTAIN ABC <t>HRP</t> kit was followed, signal was developed using <t>DAB</t> and counterstained with haematoxylin. No primary antibody was used as a control. n = 2.
    Dab Peroxidase, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 99/100, based on 935 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    CXCR4 and ACKR3 staining using IHC in breast cancer tissue. 4 μm sections from human breast cancer were stained for CXCR4 (1:40) and ACKR3 (1:100) using immunohistochemistry following no pre-treatment or EDTA antigen retrieval pre-treatment, respectively. Briefly, protocol from the VECTASTAIN ABC HRP kit was followed, signal was developed using DAB and counterstained with haematoxylin. No primary antibody was used as a control. n = 2.

    Journal: International Journal of Molecular Sciences

    Article Title: Breast Cancer: An Examination of the Potential of ACKR3 to Modify the Response of CXCR4 to CXCL12

    doi: 10.3390/ijms19113592

    Figure Lengend Snippet: CXCR4 and ACKR3 staining using IHC in breast cancer tissue. 4 μm sections from human breast cancer were stained for CXCR4 (1:40) and ACKR3 (1:100) using immunohistochemistry following no pre-treatment or EDTA antigen retrieval pre-treatment, respectively. Briefly, protocol from the VECTASTAIN ABC HRP kit was followed, signal was developed using DAB and counterstained with haematoxylin. No primary antibody was used as a control. n = 2.

    Article Snippet: Secondary and tertiary antibodies were added for 30 min before dispensing DAB on top of the samples (DAB Peroxidase (HRP) Substrate Kit, Vector labs, Burlingame, CA, USA) and counterstaining with haematoxylin.

    Techniques: Staining, Immunohistochemistry

    Immunostaining of monokine induced by interferon γ (Mig)/CXCL9, interferon-inducible protein-10 (IP-10)/CXCL10 and interferon-inducible T cell α-chemoattractant (I-TAC)/CXCL11 in bronchoalveolar lavage (BA) cells and sarcoid lungs. BAL cells were cytocentrifuged on glass slides and fixed. The slides were incubated with control rabbit serum (a), anti-Mig/CXCL9 (b), IP-10/CXCL10 (c) or I-TAC/CXCL11 antibody (d), and stained with Alexa fluor 488-conjugated goat anti-rabbit IgG (H + l) antibody (green fluorescence) and 4′,6 diamidino-2-phenylindole (DAPI; by which nuclei were counterstained) (blue). Immunostaining was visualized using a fluorescence microscope (original magnifications: × 400). Arrows point to the lymphocytes which are negative for CXCR3 ligands. Freshly frozen lung sections derived from three patients with stage II of sarcoidosis were also stained with control rabbit serum (e), anti-Mig/CXCL9 (f), IP-10/CXCL10 (g) or I-TAC/CXCL11 antibody (h) and RTU Vectastain Universal Quick Kit, and developed with diaminobenzidine (DAB) substrate (original magnifications: × 200). The insets in e, f, g and h show the staining of macrophages in the alveolar lumen (original magnifications: × 400). Scale bars = 50 µm.

    Journal: Clinical and Experimental Immunology

    Article Title: CXCL9 and 11 in patients with pulmonary sarcoidosis: a role of alveolar macrophages

    doi: 10.1111/j.1365-2249.2007.03423.x

    Figure Lengend Snippet: Immunostaining of monokine induced by interferon γ (Mig)/CXCL9, interferon-inducible protein-10 (IP-10)/CXCL10 and interferon-inducible T cell α-chemoattractant (I-TAC)/CXCL11 in bronchoalveolar lavage (BA) cells and sarcoid lungs. BAL cells were cytocentrifuged on glass slides and fixed. The slides were incubated with control rabbit serum (a), anti-Mig/CXCL9 (b), IP-10/CXCL10 (c) or I-TAC/CXCL11 antibody (d), and stained with Alexa fluor 488-conjugated goat anti-rabbit IgG (H + l) antibody (green fluorescence) and 4′,6 diamidino-2-phenylindole (DAPI; by which nuclei were counterstained) (blue). Immunostaining was visualized using a fluorescence microscope (original magnifications: × 400). Arrows point to the lymphocytes which are negative for CXCR3 ligands. Freshly frozen lung sections derived from three patients with stage II of sarcoidosis were also stained with control rabbit serum (e), anti-Mig/CXCL9 (f), IP-10/CXCL10 (g) or I-TAC/CXCL11 antibody (h) and RTU Vectastain Universal Quick Kit, and developed with diaminobenzidine (DAB) substrate (original magnifications: × 200). The insets in e, f, g and h show the staining of macrophages in the alveolar lumen (original magnifications: × 400). Scale bars = 50 µm.

    Article Snippet: Sections were developed with a diaminobenzidine substrate kit (Vector Laboratories, Inc.) and counterstained with Mayer's haematoxylin (Muto Pure Chemicals Co., Ltd, Tokyo, Japan).

    Techniques: Immunostaining, Incubation, Staining, Fluorescence, Microscopy, Derivative Assay

    Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in human breast carcinoma cells. Sections of two different breast tumours were stained and analysed as described for Figure 5. Brightfield images of the two different tumours are shown ( A,G ), with the unmixed channels for 3,3-diaminobenzidine (Ki67) ( B,H ) and SG Blue (p-ERK) ( C,I ). Composite images showing whole-cell segmentation of the tumour (cytokeratin-positive) cells are shown ( D,J ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for tumour cells ( E,K ) and for non-tumour (stromal) cells ( F,L ), with each dot representing one cell.

    Journal: Histopathology

    Article Title: Cell-based quantification of molecular biomarkers in histopathology specimens

    doi: 10.1111/j.1365-2559.2011.03878.x

    Figure Lengend Snippet: Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in human breast carcinoma cells. Sections of two different breast tumours were stained and analysed as described for Figure 5. Brightfield images of the two different tumours are shown ( A,G ), with the unmixed channels for 3,3-diaminobenzidine (Ki67) ( B,H ) and SG Blue (p-ERK) ( C,I ). Composite images showing whole-cell segmentation of the tumour (cytokeratin-positive) cells are shown ( D,J ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for tumour cells ( E,K ) and for non-tumour (stromal) cells ( F,L ), with each dot representing one cell.

    Article Snippet: ER, PR, Ki67, p-ERK and HER2 were detected by immunohistochemistry with biotinylated species-specific secondary antibodies, avidin-linked horseradish peroxidase (HRP) (ABC Kit) and 3,3-diaminobenzidine or SG Blue (Vector Laboratories) HRP chromogen substrate.

    Techniques: Immunostaining, Staining

    Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in lymphoid cells in a human breast carcinoma. A section of a breast tumour was stained sequentially with anti-p-ERK (SG Blue), anti-Ki67 [3,3-diaminobenzidine (DAB)] and anti-CK (Alexa-488) antibodies, and this was followed by haematoxylin staining, multispectral imaging (×400), and cytometric analysis. The brightfield image of a lymphoid nodule in the tumour is shown ( A ), along with the unmixed channels for DAB (Ki67) ( B ), SG Blue (p-ERK) ( C ), and Alexa-488 (cytokeratin) ( D ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for cells in the lymphoid nodule ( E ) and for tumour cells ( F ), with each dot representing one cell.

    Journal: Histopathology

    Article Title: Cell-based quantification of molecular biomarkers in histopathology specimens

    doi: 10.1111/j.1365-2559.2011.03878.x

    Figure Lengend Snippet: Duplex analysis of phospho-extracellular signal-related kinase (p-ERK) and Ki67 immunostaining in lymphoid cells in a human breast carcinoma. A section of a breast tumour was stained sequentially with anti-p-ERK (SG Blue), anti-Ki67 [3,3-diaminobenzidine (DAB)] and anti-CK (Alexa-488) antibodies, and this was followed by haematoxylin staining, multispectral imaging (×400), and cytometric analysis. The brightfield image of a lymphoid nodule in the tumour is shown ( A ), along with the unmixed channels for DAB (Ki67) ( B ), SG Blue (p-ERK) ( C ), and Alexa-488 (cytokeratin) ( D ). Scatter plots of p-ERK ( x -axis) and Ki67 ( y -axis) staining intensity are shown for cells in the lymphoid nodule ( E ) and for tumour cells ( F ), with each dot representing one cell.

    Article Snippet: ER, PR, Ki67, p-ERK and HER2 were detected by immunohistochemistry with biotinylated species-specific secondary antibodies, avidin-linked horseradish peroxidase (HRP) (ABC Kit) and 3,3-diaminobenzidine or SG Blue (Vector Laboratories) HRP chromogen substrate.

    Techniques: Immunostaining, Staining, Imaging

    Ultra-structural analysis of Sox-2 expression. A. Sox-2 immuno-gold label is observed in the nucleus (arrowheads) in ependymal cells and astrocytes in ultra-thin sections of the SVZ. Boxed areas show an ependymal cell (E) and an astrocyte (As). Ependymal layer (EL), gap layer (HL), astrocyte ribbon layer (R). Scale bar = 20 um; Scale bar in boxed areas = 1 um. a1. Magnification shows a Sox-2 + astrocyte and lack of immunoreactivity in an oligodendrocyte (O) and a microglial cell (Mi). Astrocytes are recognized by the presence of dense bundles of intermediate filaments (arrows). Scale bar = 2 um. B. Sox-2 immuno-gold staining in a group of neuroblasts (type A cells). Scale bar = 20 um. b1. Magnification of three neuroblasts showing the nuclear staining. Neuroblasts form chain-like structures with characteristic intercellular spaces (arrows). Scale bar = 2 um. C. Light microscopy image of a semi-thin section showing the distribution and characterization of cells expressing Sox-2 in the dentate gyrus. Sox-2 + cells are visualized by the nuclear dark brown DAB precipitate. The section was counterstained with toluidine blue. The line marks the boundary between the granular layer (GrL) and the hilus (H) where most labeled cells were located. Scale bar = 100 um. c1 ) Neurons in the granular layer were not Sox-2 + . Scale bar = 10 um; Insert = 1 um. D. Microphotograph of a striatal field of a control monkey showing a Sox-2 + astrocyte and unlabeled neuron (Neu) and microglial cells (Mi). Scale bar = 5 um. d1. Boxed area of the astrocyte (As) nucleus shows the gold particles (arrowheads). Scale bar = 1 um. E. A representative Sox-2 + neuron in the striatum of a control monkey. Scale bar = 2 um. e1. Boxed area shows deposition of gold particles over the nucleus (arrowheads) close to a synapse (arrow), which identifies this cell as a neuron. Scale bar = 1 um. F. Sox-2 + astrocyte in the substantia nigra pars compacta. Scale bar = 10 µm. f1. Boxed area shows a dense bundle of intermediate filaments (arrows) that characterizes astrocytes. Arrowheads indicate gold particles in the nucleus. Scale bar = 1 µm. Abbreviations: subventricular zone: (SVZ); 3′, 3′-diaminobenzidine: (DAB).

    Journal: PLoS ONE

    Article Title: Sox-2 Positive Neural Progenitors in the Primate Striatum Undergo Dynamic Changes after Dopamine Denervation

    doi: 10.1371/journal.pone.0066377

    Figure Lengend Snippet: Ultra-structural analysis of Sox-2 expression. A. Sox-2 immuno-gold label is observed in the nucleus (arrowheads) in ependymal cells and astrocytes in ultra-thin sections of the SVZ. Boxed areas show an ependymal cell (E) and an astrocyte (As). Ependymal layer (EL), gap layer (HL), astrocyte ribbon layer (R). Scale bar = 20 um; Scale bar in boxed areas = 1 um. a1. Magnification shows a Sox-2 + astrocyte and lack of immunoreactivity in an oligodendrocyte (O) and a microglial cell (Mi). Astrocytes are recognized by the presence of dense bundles of intermediate filaments (arrows). Scale bar = 2 um. B. Sox-2 immuno-gold staining in a group of neuroblasts (type A cells). Scale bar = 20 um. b1. Magnification of three neuroblasts showing the nuclear staining. Neuroblasts form chain-like structures with characteristic intercellular spaces (arrows). Scale bar = 2 um. C. Light microscopy image of a semi-thin section showing the distribution and characterization of cells expressing Sox-2 in the dentate gyrus. Sox-2 + cells are visualized by the nuclear dark brown DAB precipitate. The section was counterstained with toluidine blue. The line marks the boundary between the granular layer (GrL) and the hilus (H) where most labeled cells were located. Scale bar = 100 um. c1 ) Neurons in the granular layer were not Sox-2 + . Scale bar = 10 um; Insert = 1 um. D. Microphotograph of a striatal field of a control monkey showing a Sox-2 + astrocyte and unlabeled neuron (Neu) and microglial cells (Mi). Scale bar = 5 um. d1. Boxed area of the astrocyte (As) nucleus shows the gold particles (arrowheads). Scale bar = 1 um. E. A representative Sox-2 + neuron in the striatum of a control monkey. Scale bar = 2 um. e1. Boxed area shows deposition of gold particles over the nucleus (arrowheads) close to a synapse (arrow), which identifies this cell as a neuron. Scale bar = 1 um. F. Sox-2 + astrocyte in the substantia nigra pars compacta. Scale bar = 10 µm. f1. Boxed area shows a dense bundle of intermediate filaments (arrows) that characterizes astrocytes. Arrowheads indicate gold particles in the nucleus. Scale bar = 1 µm. Abbreviations: subventricular zone: (SVZ); 3′, 3′-diaminobenzidine: (DAB).

    Article Snippet: The chromogen solution was 3′, 3′-diaminobenzidine (DAB) with H2 O2 (DAB kit, Vector Laboratories).

    Techniques: Expressing, Staining, Light Microscopy, Labeling