nickel enhanced dab  (Vector Laboratories)


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    Name:
    Biotinylated Goat Anti Rabbit IgG Antibody
    Description:
    Biotinylated Goat Anit Rabbit IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Goat Anti Rabbit IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    Catalog Number:
    ba-1000
    Price:
    None
    Host:
    Goat
    Size:
    1 5 mg
    Category:
    Antibodies
    Reactivity:
    Rabbit
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    Structured Review

    Vector Laboratories nickel enhanced dab
    Biotinylated Goat Anti Rabbit IgG Antibody
    Biotinylated Goat Anit Rabbit IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Goat Anti Rabbit IgG H L is supplied in liquid format With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    https://www.bioz.com/result/nickel enhanced dab/product/Vector Laboratories
    Average 88 stars, based on 965 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 "Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice"

    Article Title: Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00560.2017

    Deficits in proliferation and survival of the cells of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were harvested on PND14 for lung immunohistochemistry studies. Representative lung sections showing Ki67-stained cells from mice treated on PND3 with PBS ( A ) and L10 ( B ) and on PND7 with PBS ( C ) and L10 ( D ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated on PND3 with PBS ( E ) and L10 ( F ) and on PND7 with PBS ( G ) and L10 ( H ). Quantification of Ki67-positive (percentage) ( I ) and cleaved caspase 3-positive (number per high power field) ( J ) lung cells. Values are presented as means ± SD ( n = 6/group). Significant differences between age-matched PBS- and L10-exposed animals are indicated by *** P
    Figure Legend Snippet: Deficits in proliferation and survival of the cells of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were harvested on PND14 for lung immunohistochemistry studies. Representative lung sections showing Ki67-stained cells from mice treated on PND3 with PBS ( A ) and L10 ( B ) and on PND7 with PBS ( C ) and L10 ( D ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated on PND3 with PBS ( E ) and L10 ( F ) and on PND7 with PBS ( G ) and L10 ( H ). Quantification of Ki67-positive (percentage) ( I ) and cleaved caspase 3-positive (number per high power field) ( J ) lung cells. Values are presented as means ± SD ( n = 6/group). Significant differences between age-matched PBS- and L10-exposed animals are indicated by *** P

    Techniques Used: Mouse Assay, Injection, Immunohistochemistry, Staining

    Effect of chronic LPS exposure on lung cell proliferation and apoptosis. Newborn mice were treated intraperitoneally with 10 mg/kg of LPS (L10) or a vehicle control (PBS) on postnatal days (PNDs) 3–5 , and the lung tissues were harvested for immunohistochemistry studies on PND7. Representative lung sections showing Ki67-stained cells from mice treated with PBS ( A ) and L10 ( B ). Quantification of the percentage of Ki67-positive lung cells ( C ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated with PBS ( D ) and L10 ( E ). Quantification of cleaved caspase 3-positive lung cells ( F ). HPF, high-power field. Values are presented as means ± SD ( n = 6/group). Significant differences between PBS- and L10-treated animals are indicated by *** P
    Figure Legend Snippet: Effect of chronic LPS exposure on lung cell proliferation and apoptosis. Newborn mice were treated intraperitoneally with 10 mg/kg of LPS (L10) or a vehicle control (PBS) on postnatal days (PNDs) 3–5 , and the lung tissues were harvested for immunohistochemistry studies on PND7. Representative lung sections showing Ki67-stained cells from mice treated with PBS ( A ) and L10 ( B ). Quantification of the percentage of Ki67-positive lung cells ( C ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated with PBS ( D ) and L10 ( E ). Quantification of cleaved caspase 3-positive lung cells ( F ). HPF, high-power field. Values are presented as means ± SD ( n = 6/group). Significant differences between PBS- and L10-treated animals are indicated by *** P

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining

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

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

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086892

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

    Techniques Used: Expressing, Microarray

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

    Techniques Used: Microarray, Construct, Transformation Assay, Expressing

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

    Techniques Used: Expressing, Microarray

    4) Product Images from "Early life trauma increases threat response of peri-weaning rats, reduction of axo-somatic synapses formed by parvalbumin cells and perineuronal net in the basolateral nucleus of amygdala"

    Article Title: Early life trauma increases threat response of peri-weaning rats, reduction of axo-somatic synapses formed by parvalbumin cells and perineuronal net in the basolateral nucleus of amygdala

    Journal: The Journal of comparative neurology

    doi: 10.1002/cne.24522

    Specificity of WFA labeling of PNN in the BLA of weanling rats. Panel a: HRP-DAB was used to detect WFA labeling. Specificity of WFA labeling was confirmed by reduction of WFA labeling within the brains injected with the enzymes, chondroitinase-ABC and hyaluronidase, known to dissolve proteoglycans of the PNN and surrounding neuropil. The region infused with chondroitinase-ABC and hyaluronidase exhibited complete absence of the HRP-DAB reaction product, while PNN labeling remained intensely labeled in the surrounding regions (e.g., dorsal hippocampus and reticular thalamus) (scale bar= 200 µm). Panel b : The pink contour indicates the boundary of the BLA (scale bar = 200 µm). Panel c : The lower right panel shows detail of the enzyme-infused region (scale bar = 50 µm). All panels were taken at a magnification of 10X.
    Figure Legend Snippet: Specificity of WFA labeling of PNN in the BLA of weanling rats. Panel a: HRP-DAB was used to detect WFA labeling. Specificity of WFA labeling was confirmed by reduction of WFA labeling within the brains injected with the enzymes, chondroitinase-ABC and hyaluronidase, known to dissolve proteoglycans of the PNN and surrounding neuropil. The region infused with chondroitinase-ABC and hyaluronidase exhibited complete absence of the HRP-DAB reaction product, while PNN labeling remained intensely labeled in the surrounding regions (e.g., dorsal hippocampus and reticular thalamus) (scale bar= 200 µm). Panel b : The pink contour indicates the boundary of the BLA (scale bar = 200 µm). Panel c : The lower right panel shows detail of the enzyme-infused region (scale bar = 50 µm). All panels were taken at a magnification of 10X.

    Techniques Used: Labeling, Injection

    5) Product Images from "Modulation of Sodium/Iodide Symporter Expression in the Salivary Gland"

    Article Title: Modulation of Sodium/Iodide Symporter Expression in the Salivary Gland

    Journal: Thyroid

    doi: 10.1089/thy.2012.0571

    Sodium/iodide symporter (NIS) expression is primarily restricted to striated ducts in human salivary glands. (A) NIS is expressed at a high level along basolateral membranes of striated duct cells (S), with fewer intercalated (rectangle) and excretory (E) duct cells demonstrating lower NIS levels. Acinar (A) cells do not express NIS. Anti-human NIS antibody, avidin-biotin complex, DAB chromogen, hematoxylin counterstain, bar=25 μm. (B) Distribution and intensity of NIS immunoreactivity among intercalated (ID), striated (SD), and excretory (ED) ductal cells in normal human submandibular, parotid, and minor salivary glands. NIS expression is greater and more frequent in striated ducts, regardless of gland type.
    Figure Legend Snippet: Sodium/iodide symporter (NIS) expression is primarily restricted to striated ducts in human salivary glands. (A) NIS is expressed at a high level along basolateral membranes of striated duct cells (S), with fewer intercalated (rectangle) and excretory (E) duct cells demonstrating lower NIS levels. Acinar (A) cells do not express NIS. Anti-human NIS antibody, avidin-biotin complex, DAB chromogen, hematoxylin counterstain, bar=25 μm. (B) Distribution and intensity of NIS immunoreactivity among intercalated (ID), striated (SD), and excretory (ED) ductal cells in normal human submandibular, parotid, and minor salivary glands. NIS expression is greater and more frequent in striated ducts, regardless of gland type.

    Techniques Used: Expressing, Avidin-Biotin Assay

    NIS expression is reduced in inflamed and neoplastic human salivary glands. (A) NIS expression in striated ducts is decreased in inflammatory salivary glands, as well as in benign and malignant neoplasms of ductal origin. (B) NIS immunoreactivity in inflamed (i, ii) and neoplastic (iii, iv) salivary tissue. NIS expression is variably decreased in striated ducts (S) of chronically inflamed salivary glands (i, ii) , particularly in ducts demonstrating goblet metaplasia (arrows). Neoplastic cells of Warthin's tumor (iii) and mucoepidermoid carcinoma (iv) demonstrate lower NIS level, with immunostaining intensity of 2+ and 1+, respectively. Anti-human NIS antibody, avidin-biotin complex, DAB chromogen, hematoxylin counterstain, bars=25 μm. F, periductal fibrosis; L, lymphocytic inflammation.
    Figure Legend Snippet: NIS expression is reduced in inflamed and neoplastic human salivary glands. (A) NIS expression in striated ducts is decreased in inflammatory salivary glands, as well as in benign and malignant neoplasms of ductal origin. (B) NIS immunoreactivity in inflamed (i, ii) and neoplastic (iii, iv) salivary tissue. NIS expression is variably decreased in striated ducts (S) of chronically inflamed salivary glands (i, ii) , particularly in ducts demonstrating goblet metaplasia (arrows). Neoplastic cells of Warthin's tumor (iii) and mucoepidermoid carcinoma (iv) demonstrate lower NIS level, with immunostaining intensity of 2+ and 1+, respectively. Anti-human NIS antibody, avidin-biotin complex, DAB chromogen, hematoxylin counterstain, bars=25 μm. F, periductal fibrosis; L, lymphocytic inflammation.

    Techniques Used: Expressing, Immunostaining, Avidin-Biotin Assay

    6) Product Images from "The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis"

    Article Title: The necroptotic cell death pathway operates in megakaryocytes, but not in platelet synthesis

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-021-03418-z

    Morphology of megakaryocytes in response to TNF-induced necroptosis and apoptosis. a Representative time-lapse confocal images from WT BM-derived cultured megakaryocytes treated with TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), QVD-OPh (50 µM, Q), Necrostatin-1 (50 µM, N), and ABT-737 (5 µM). Megakaryocytes were labelled with CD41 conjugated to Alexa-488 (green) and PI (propidium iodide, red), Bar 50 µm. Cells were maintained on the stage within a humidified chamber at 37 °C and 5% CO 2 for 7 h. Images were acquired on a Ziess live cell Axio inverted modular microscope for live cell imaging every 10 min driven by AxioVision v4.8 software. b Representative time-lapse confocal images from WT and Mlkl −/− BM-derived cultured megakaryocytes treated with the indicated agents: TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), IDN-6556 (5 µM, I), Necrostatin-1 (50 µM, N) at the indicated time points (hours) at 37 °C, images acquired as in ( a ). c Quantification of PI from representative time-lapse video microscopy at the indicated time points (hours). PI fluorescence intensity was quantified using a custom macro in Fiji software. n = 3 independent experiments, data presented as mean ± SEM, One-way ANOVA with the Bonferroni multiple comparison test, ** P
    Figure Legend Snippet: Morphology of megakaryocytes in response to TNF-induced necroptosis and apoptosis. a Representative time-lapse confocal images from WT BM-derived cultured megakaryocytes treated with TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), QVD-OPh (50 µM, Q), Necrostatin-1 (50 µM, N), and ABT-737 (5 µM). Megakaryocytes were labelled with CD41 conjugated to Alexa-488 (green) and PI (propidium iodide, red), Bar 50 µm. Cells were maintained on the stage within a humidified chamber at 37 °C and 5% CO 2 for 7 h. Images were acquired on a Ziess live cell Axio inverted modular microscope for live cell imaging every 10 min driven by AxioVision v4.8 software. b Representative time-lapse confocal images from WT and Mlkl −/− BM-derived cultured megakaryocytes treated with the indicated agents: TNF (100 ng/ml, T), Smac mimetic (0.5 µM, S), IDN-6556 (5 µM, I), Necrostatin-1 (50 µM, N) at the indicated time points (hours) at 37 °C, images acquired as in ( a ). c Quantification of PI from representative time-lapse video microscopy at the indicated time points (hours). PI fluorescence intensity was quantified using a custom macro in Fiji software. n = 3 independent experiments, data presented as mean ± SEM, One-way ANOVA with the Bonferroni multiple comparison test, ** P

    Techniques Used: Derivative Assay, Cell Culture, Microscopy, Live Cell Imaging, Software, Fluorescence

    7) 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

    8) 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:

    9) 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

    10) Product Images from "Fas and Fas Ligand Are Up-Regulated in Pulmonary Edema Fluid and Lung Tissue of Patients with Acute Lung Injury and the Acute Respiratory Distress Syndrome"

    Article Title: Fas and Fas Ligand Are Up-Regulated in Pulmonary Edema Fluid and Lung Tissue of Patients with Acute Lung Injury and the Acute Respiratory Distress Syndrome

    Journal: The American Journal of Pathology

    doi:

    Localization of markers of apoptosis in lung tissue sections from patients. The left column shows lung tissue sections from a patient who died with ALI or ARDS [identified as “(+) ARDS”]. The right column shows lung tissue sections from a patient who died without pulmonary disease [identified as “(−) ARDS”]. a to f: Tissue sections that are counterstained with hematoxylin. g to j: Tissue sections that were imaged using differential interference contrast optics, without counterstain, because the epithelial cell immunostain reaction product was subtle. The rows of pictures are matched for one marker of apoptosis: TUNEL ( a and b ), caspase-3 ( c and d ), Bax ( e and f ), Bcl II ( g and h ), and p53 ( i and j ). Cells lining, and in, the alveolar walls demonstrate more TUNEL-labeled nuclei ( arrow ), caspase-3-labeled cytoplasm ( arrow ), Bax-labeled cytoplasm ( arrow ), and p53-labeled cytoplasm ( arrow ) in the tissue sections from the patients who died with ALI or ARDS compared to the patient who died without pulmonary disease. On the other hand, Bcl II-labeled cells lining, and in, the alveolar walls are more prominent in the tissue sections from the patient who died without pulmonary disease compared to the patient who died with ALI or ARDS, as expected. All of the panels are the same magnification.
    Figure Legend Snippet: Localization of markers of apoptosis in lung tissue sections from patients. The left column shows lung tissue sections from a patient who died with ALI or ARDS [identified as “(+) ARDS”]. The right column shows lung tissue sections from a patient who died without pulmonary disease [identified as “(−) ARDS”]. a to f: Tissue sections that are counterstained with hematoxylin. g to j: Tissue sections that were imaged using differential interference contrast optics, without counterstain, because the epithelial cell immunostain reaction product was subtle. The rows of pictures are matched for one marker of apoptosis: TUNEL ( a and b ), caspase-3 ( c and d ), Bax ( e and f ), Bcl II ( g and h ), and p53 ( i and j ). Cells lining, and in, the alveolar walls demonstrate more TUNEL-labeled nuclei ( arrow ), caspase-3-labeled cytoplasm ( arrow ), Bax-labeled cytoplasm ( arrow ), and p53-labeled cytoplasm ( arrow ) in the tissue sections from the patients who died with ALI or ARDS compared to the patient who died without pulmonary disease. On the other hand, Bcl II-labeled cells lining, and in, the alveolar walls are more prominent in the tissue sections from the patient who died without pulmonary disease compared to the patient who died with ALI or ARDS, as expected. All of the panels are the same magnification.

    Techniques Used: Marker, TUNEL Assay, Labeling

    11) Product Images from "Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?"

    Article Title: Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25203

    Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.
    Figure Legend Snippet: Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.

    Techniques Used: Neutralization

    12) 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

    13) Product Images from "Runx2 Expression in Smooth Muscle Cells Is Required for Arterial Medial Calcification in Mice"

    Article Title: Runx2 Expression in Smooth Muscle Cells Is Required for Arterial Medial Calcification in Mice

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.03.020

    Smooth muscle cell (SMC)-specific deletion of Runx2 prevents SMC osteochondrogenic phenotype change. Runx2 f/f ( A, B, D, E, G, H, J, K, M , and N ) and Runx2 ΔSM ( C, F, I, L , and O ) mice were treated with either vitamin D (VitD) ( B, C, E, F, H, I, K, L, N , and O ) or vehicle control ( A, D, G, J , and M . Abdominal aortas were collected to visualize calcium deposition through Alizarin Red S stain ( A–C ), as well as hematoxylin and eosin (H E) ( D–F ), Runx2 (brown nuclear stain) ( G–I ), SMC marker protein, SM22α (brown stain) ( J–L ), and osteopontin (OPN) (brown stain) ( M–O , double arrows ). Dashed lines in G–I designate external elastic lamina. Note the bright red calcium stain in the aortic media of Runx2 f/f mice challenged with vitamin D ( B ) and the presence of Runx2-positive cells in the media layer around the calcification site ( H , arrowheads ), but not in vehicle-injected ( A ) or Runx2 ΔSM vessels ( C ). Note also the concomitant loss of SM22α at the calcification site ( K , arrows ).
    Figure Legend Snippet: Smooth muscle cell (SMC)-specific deletion of Runx2 prevents SMC osteochondrogenic phenotype change. Runx2 f/f ( A, B, D, E, G, H, J, K, M , and N ) and Runx2 ΔSM ( C, F, I, L , and O ) mice were treated with either vitamin D (VitD) ( B, C, E, F, H, I, K, L, N , and O ) or vehicle control ( A, D, G, J , and M . Abdominal aortas were collected to visualize calcium deposition through Alizarin Red S stain ( A–C ), as well as hematoxylin and eosin (H E) ( D–F ), Runx2 (brown nuclear stain) ( G–I ), SMC marker protein, SM22α (brown stain) ( J–L ), and osteopontin (OPN) (brown stain) ( M–O , double arrows ). Dashed lines in G–I designate external elastic lamina. Note the bright red calcium stain in the aortic media of Runx2 f/f mice challenged with vitamin D ( B ) and the presence of Runx2-positive cells in the media layer around the calcification site ( H , arrowheads ), but not in vehicle-injected ( A ) or Runx2 ΔSM vessels ( C ). Note also the concomitant loss of SM22α at the calcification site ( K , arrows ).

    Techniques Used: Mouse Assay, Staining, Marker, Injection

    14) Product Images from "Modulation of lethal HPAIV H5N8 clade 2.3.4.4B infection in AIV pre-exposed mallards"

    Article Title: Modulation of lethal HPAIV H5N8 clade 2.3.4.4B infection in AIV pre-exposed mallards

    Journal: Emerging Microbes & Infections

    doi: 10.1080/22221751.2020.1713706

    Macroscopic findings in the livers of experimentally H5N8B-infected ducks. (A) Seronegative Pekin duck, H5N8B-infected, euthanized 9 dpi due to neurological symptoms, liver. Macroscopically normal, brown-red, acutely-angled liver without immunohistochemically-detectable hepatocellular influenza A virus matrix protein. (B) Pekin contact duckling in the mallard group, died 4 dpc, liver. Swollen, brick-red-colored, friable liver with rounded edges, interpreted as severe, acute, diffuse, necrotizing hepatitis with immunohistochemically-detectable hepatocellular influenza A virus matrix protein. (C) Seropositive mallard, H5N8B-infected, clinically normal, 14 dpi, liver. Swollen, beige, greasy liver with rounded edges, interpreted as moderate, acute, diffuse hepatocellular lipidosis (background pathology) without immunohistochemically detectable hepatocellular influenza A virus matrixprotein.
    Figure Legend Snippet: Macroscopic findings in the livers of experimentally H5N8B-infected ducks. (A) Seronegative Pekin duck, H5N8B-infected, euthanized 9 dpi due to neurological symptoms, liver. Macroscopically normal, brown-red, acutely-angled liver without immunohistochemically-detectable hepatocellular influenza A virus matrix protein. (B) Pekin contact duckling in the mallard group, died 4 dpc, liver. Swollen, brick-red-colored, friable liver with rounded edges, interpreted as severe, acute, diffuse, necrotizing hepatitis with immunohistochemically-detectable hepatocellular influenza A virus matrix protein. (C) Seropositive mallard, H5N8B-infected, clinically normal, 14 dpi, liver. Swollen, beige, greasy liver with rounded edges, interpreted as moderate, acute, diffuse hepatocellular lipidosis (background pathology) without immunohistochemically detectable hepatocellular influenza A virus matrixprotein.

    Techniques Used: Infection

    Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.
    Figure Legend Snippet: Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.

    Techniques Used: Infection, Lysis, Immunohistochemistry, Avidin-Biotin Assay

    15) Product Images from "The Importance of Titrating Antibodies for Immunocytochemical Methods"

    Article Title: The Importance of Titrating Antibodies for Immunocytochemical Methods

    Journal: Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.]

    doi: 10.1002/cpns.1

    The ABC peroxidase technique. This approach is similar to that shown in , but substitutes a biotinylated peroxidase. When incubated with a substrate such as DAB (lower left) or NiDAB (lower right), the colored insoluble product is deposited
    Figure Legend Snippet: The ABC peroxidase technique. This approach is similar to that shown in , but substitutes a biotinylated peroxidase. When incubated with a substrate such as DAB (lower left) or NiDAB (lower right), the colored insoluble product is deposited

    Techniques Used: Incubation

    TSA-amplified fluorescence using biotinylated tyramine and streptavidin fluorophore. Note the greatly increased number of fluorescent molecules compared with that seen for either fluorophore-tagged secondaries or ABC streptavidin methods. P = peroxidase.
    Figure Legend Snippet: TSA-amplified fluorescence using biotinylated tyramine and streptavidin fluorophore. Note the greatly increased number of fluorescent molecules compared with that seen for either fluorophore-tagged secondaries or ABC streptavidin methods. P = peroxidase.

    Techniques Used: Amplification, Fluorescence

    16) Product Images from "Interferon regulatory factor 2 protects mice from lethal viral neuroinvasion"

    Article Title: Interferon regulatory factor 2 protects mice from lethal viral neuroinvasion

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20160303

    B cells and virus-specific IgG level are significantly reduced in Irf2 −/− mouse brains. (A) Brain slides from five Irf2 −/− moribund mice and six SVN-infected WT mice with high CNS viral titers from Fig. 2 D were stained with antibodies specific for CD3 (T cell), B220 (B cell), granzyme (cytotoxic T and NK cells), and Mac2 (macrophage). Immune cell infiltrates were semiquantitatively scored using a scale of zero to four as follows: 0, absent; 1, minimal; 2, mild; 3, moderate; and 4, marked. (B) Images that consistently sample eight different regions in the brains (Fig. S2) of WT and Irf2 −/− mice were taken, and the B cell staining present in those images was quantified by ImageJ (measurement of number of positive pixels) or by eye (number of B cells). The p-values were determined by the unpaired, two-tailed Student’s t test (*, P
    Figure Legend Snippet: B cells and virus-specific IgG level are significantly reduced in Irf2 −/− mouse brains. (A) Brain slides from five Irf2 −/− moribund mice and six SVN-infected WT mice with high CNS viral titers from Fig. 2 D were stained with antibodies specific for CD3 (T cell), B220 (B cell), granzyme (cytotoxic T and NK cells), and Mac2 (macrophage). Immune cell infiltrates were semiquantitatively scored using a scale of zero to four as follows: 0, absent; 1, minimal; 2, mild; 3, moderate; and 4, marked. (B) Images that consistently sample eight different regions in the brains (Fig. S2) of WT and Irf2 −/− mice were taken, and the B cell staining present in those images was quantified by ImageJ (measurement of number of positive pixels) or by eye (number of B cells). The p-values were determined by the unpaired, two-tailed Student’s t test (*, P

    Techniques Used: Mouse Assay, Infection, Staining, Two Tailed Test

    17) Product Images from "β-Catenin C-terminal signals suppress p53 and are essential for artery formation"

    Article Title: β-Catenin C-terminal signals suppress p53 and are essential for artery formation

    Journal: Nature Communications

    doi: 10.1038/ncomms12389

    β-Catenin promotes SMC proliferation and survival during artery formation. ( a ) Immunohistochemistry (IHC) for the endothelial marker CD31. Arrowheads indicate the endothelial layer. Scale bar, 50 μm. ( b ) IHC for SMC markers, Sm22α (brown; scale bar, 25 μm) and α-SMA (red; scale bar, 50 μm). Arrowheads delimit the vessel wall. Arrows indicate scattered SMCs. ( c ) Immunostaining of PDAs for the mitotic marker pHH3 and α-SMA at E10.5. Arrowheads indicate pHH3 + SMCs. Scale bar, 20 μm. ( d ) Quantification of pHH3 + SMCs in the wall of PDAs. ** P
    Figure Legend Snippet: β-Catenin promotes SMC proliferation and survival during artery formation. ( a ) Immunohistochemistry (IHC) for the endothelial marker CD31. Arrowheads indicate the endothelial layer. Scale bar, 50 μm. ( b ) IHC for SMC markers, Sm22α (brown; scale bar, 25 μm) and α-SMA (red; scale bar, 50 μm). Arrowheads delimit the vessel wall. Arrows indicate scattered SMCs. ( c ) Immunostaining of PDAs for the mitotic marker pHH3 and α-SMA at E10.5. Arrowheads indicate pHH3 + SMCs. Scale bar, 20 μm. ( d ) Quantification of pHH3 + SMCs in the wall of PDAs. ** P

    Techniques Used: Immunohistochemistry, Marker, Immunostaining

    18) Product Images from "Inhibition of FLT1 ameliorates muscular dystrophy phenotype by increased vasculature in a mouse model of Duchenne muscular dystrophy"

    Article Title: Inhibition of FLT1 ameliorates muscular dystrophy phenotype by increased vasculature in a mouse model of Duchenne muscular dystrophy

    Journal: PLoS Genetics

    doi: 10.1371/journal.pgen.1008468

    Postnatal deletion of Flt1 can increase capillary density in skeletal muscle. (A) Experimental scheme for assessing angiogenic response from conditional Flt1 deletion. (B) Representative images of CD31-stained cryosections from the TA muscle from Flt1 LoxP/LoxP ( Flt1 +/+ ) and CAG CreERTM : Flt1 LoxP/LoxP ( Flt1 Δ/Δ ) mice following tamoxifen (TMX) induction. Scale bar indicates 100 μm. (C) Increase in capillary density is dependent on CAG CreERTM as well as TMX induction in the Flt1 LoxP/LoxP background. (D) Increase in capillary density is rapid and sustained following TMX induction in Flt1 Δ/Δ mice more than 6 months following induction.
    Figure Legend Snippet: Postnatal deletion of Flt1 can increase capillary density in skeletal muscle. (A) Experimental scheme for assessing angiogenic response from conditional Flt1 deletion. (B) Representative images of CD31-stained cryosections from the TA muscle from Flt1 LoxP/LoxP ( Flt1 +/+ ) and CAG CreERTM : Flt1 LoxP/LoxP ( Flt1 Δ/Δ ) mice following tamoxifen (TMX) induction. Scale bar indicates 100 μm. (C) Increase in capillary density is dependent on CAG CreERTM as well as TMX induction in the Flt1 LoxP/LoxP background. (D) Increase in capillary density is rapid and sustained following TMX induction in Flt1 Δ/Δ mice more than 6 months following induction.

    Techniques Used: Staining, Mouse Assay

    Systemic anti-FLT1 peptide improves skeletal muscle pathology in mdx mice. (A) Experimental scheme for treatment of mdx mice with systemic treatment using anti-FLT1 peptide. (B) Representative images of (top) CD31, (middle) gross whole mount and cryosections for EBD, (bottom) Sirius red staining of diaphragm in mdx mice treated with anti-FLT1 peptide. Scale bars indicate 100 μm. (C) Anti-FLT1 peptide injection increases capillary density in the mdx mouse muscle at high dose. (D) Anti-FLT1 peptide injection is sufficient to increase skeletal muscle perfusion in mdx mice at high dose. (E) Anti-FLT1 peptide injection decreases EBD+ area in the mdx mouse muscle at high dose. (F) Anti-FLT1 peptide injection decreases fibrotic area in the mdx mouse muscle at high dose. (G) Grip strength is improved by anti-FLT1 peptide injection at high dose in mdx mice normalized to body weight.
    Figure Legend Snippet: Systemic anti-FLT1 peptide improves skeletal muscle pathology in mdx mice. (A) Experimental scheme for treatment of mdx mice with systemic treatment using anti-FLT1 peptide. (B) Representative images of (top) CD31, (middle) gross whole mount and cryosections for EBD, (bottom) Sirius red staining of diaphragm in mdx mice treated with anti-FLT1 peptide. Scale bars indicate 100 μm. (C) Anti-FLT1 peptide injection increases capillary density in the mdx mouse muscle at high dose. (D) Anti-FLT1 peptide injection is sufficient to increase skeletal muscle perfusion in mdx mice at high dose. (E) Anti-FLT1 peptide injection decreases EBD+ area in the mdx mouse muscle at high dose. (F) Anti-FLT1 peptide injection decreases fibrotic area in the mdx mouse muscle at high dose. (G) Grip strength is improved by anti-FLT1 peptide injection at high dose in mdx mice normalized to body weight.

    Techniques Used: Mouse Assay, Staining, Injection

    MAbs against FLT1 for the treatment of muscular dystrophy in the mdx mice. (A) Experimental scheme for systemic treatment of mdx mice with systemic injection of anti-FLT1 antibody. (B) Representative images of (top) CD31, (middle) EBD and (bottom) Sirius red staining of diaphragm in mdx mice treated with anti-FLT1 antibody. Scale bars indicate 100 μm. (C) Capillary destiny is increased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (D) MAB0702 but not EWC injection is sufficient to increase skeletal muscle perfusion in mdx mice. (E) EBD+ area is decreased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (F) Fibrotic area is decreased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (G) Calcification is decreased in both MAB0702 and EWC treated mice in mdx mice compared with isotype control.
    Figure Legend Snippet: MAbs against FLT1 for the treatment of muscular dystrophy in the mdx mice. (A) Experimental scheme for systemic treatment of mdx mice with systemic injection of anti-FLT1 antibody. (B) Representative images of (top) CD31, (middle) EBD and (bottom) Sirius red staining of diaphragm in mdx mice treated with anti-FLT1 antibody. Scale bars indicate 100 μm. (C) Capillary destiny is increased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (D) MAB0702 but not EWC injection is sufficient to increase skeletal muscle perfusion in mdx mice. (E) EBD+ area is decreased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (F) Fibrotic area is decreased in MAB0702 treated mice but not EWC in mdx mice compared with isotype control. (G) Calcification is decreased in both MAB0702 and EWC treated mice in mdx mice compared with isotype control.

    Techniques Used: Mouse Assay, Injection, Staining

    Increased angiogenesis is accompanied by worsened muscle pathology in the mdx : Flt1 Δ/Δ mice. (A) Representative images of CD31-stained TA muscle, Evans blue dye (EBD) to measure acute damage in diaphragm and Sirius red staining to measure fibrosis in diaphragm. Scale bars indicate 100 μm. (B) Capillary density is increased in in the TA muscle mdx : Flt1 Δ/Δ mice. (C) Skeletal muscle perfusion is increased in the TA muscle in mdx : Flt1 Δ/Δ mice. (D) mdx : Flt1 Δ/Δ mice show no difference in acute damage as judged by EBD. (E) mdx : Flt1 Δ/Δ mice show increased in fibrosis as evaluated by Sirius red staining. (F) mdx : Flt1 Δ/Δ mice show no difference in the grip strength normalized to body weight.
    Figure Legend Snippet: Increased angiogenesis is accompanied by worsened muscle pathology in the mdx : Flt1 Δ/Δ mice. (A) Representative images of CD31-stained TA muscle, Evans blue dye (EBD) to measure acute damage in diaphragm and Sirius red staining to measure fibrosis in diaphragm. Scale bars indicate 100 μm. (B) Capillary density is increased in in the TA muscle mdx : Flt1 Δ/Δ mice. (C) Skeletal muscle perfusion is increased in the TA muscle in mdx : Flt1 Δ/Δ mice. (D) mdx : Flt1 Δ/Δ mice show no difference in acute damage as judged by EBD. (E) mdx : Flt1 Δ/Δ mice show increased in fibrosis as evaluated by Sirius red staining. (F) mdx : Flt1 Δ/Δ mice show no difference in the grip strength normalized to body weight.

    Techniques Used: Mouse Assay, Staining

    Endothelial cell-specific conditional deletion of Flt1 in mdx : Cdh5-Flt1 Δ/Δ mice improve capillary density and muscle phenotype. (A) Experimental scheme for assessing angiogenic response from conditional Flt1 deletion. Cdh5-Flt1 +/+ and mdx : Cdh5-Flt1 +/+ mice were used as wild-type and mdx controls, respectively. (B) Representative images of CD31 staining for capillary density, Sirius red staining to measure fibrosis and EBD to measure acute damage in the diaphragm. Scale bars indicate 100 μm. (C) Endothelial cell specific conditional deletion of Flt1 ( mdx : Cdh5-Flt1 Δ/Δ ) is sufficient to increase the capillary density in diaphragm. (D) Endothelial cell specific conditional deletion of Flt1 ( mdx : Cdh5-Flt1 Δ/Δ ) is sufficient to increase skeletal muscle perfusion. (E) Acute damage as judged by EBD is reduced in mdx : Cdh5-Flt1 Δ/Δ mouse muscle. (F) Fibrosis is reduced in mdx : Cdh5-Flt1 Δ/Δ mouse muscle as evaluated by Sirius red staining.
    Figure Legend Snippet: Endothelial cell-specific conditional deletion of Flt1 in mdx : Cdh5-Flt1 Δ/Δ mice improve capillary density and muscle phenotype. (A) Experimental scheme for assessing angiogenic response from conditional Flt1 deletion. Cdh5-Flt1 +/+ and mdx : Cdh5-Flt1 +/+ mice were used as wild-type and mdx controls, respectively. (B) Representative images of CD31 staining for capillary density, Sirius red staining to measure fibrosis and EBD to measure acute damage in the diaphragm. Scale bars indicate 100 μm. (C) Endothelial cell specific conditional deletion of Flt1 ( mdx : Cdh5-Flt1 Δ/Δ ) is sufficient to increase the capillary density in diaphragm. (D) Endothelial cell specific conditional deletion of Flt1 ( mdx : Cdh5-Flt1 Δ/Δ ) is sufficient to increase skeletal muscle perfusion. (E) Acute damage as judged by EBD is reduced in mdx : Cdh5-Flt1 Δ/Δ mouse muscle. (F) Fibrosis is reduced in mdx : Cdh5-Flt1 Δ/Δ mouse muscle as evaluated by Sirius red staining.

    Techniques Used: Mouse Assay, Staining

    19) Product Images from "Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?"

    Article Title: Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25203

    CTGF, HIF-1, IGF-1 and IGF-1R are expressed in the majority of tumors (A) . Their expression tended to be more frequent in TCs, although this was significant only for HIF-1 (B) .
    Figure Legend Snippet: CTGF, HIF-1, IGF-1 and IGF-1R are expressed in the majority of tumors (A) . Their expression tended to be more frequent in TCs, although this was significant only for HIF-1 (B) .

    Techniques Used: Expressing

    Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.
    Figure Legend Snippet: Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.

    Techniques Used: Neutralization

    20) Product Images from "Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?"

    Article Title: Can insulin-like growth factor 1 (IGF-1), IGF-1 receptor connective tissue growth factor and Ki-67 labelling index have a prognostic role in pulmonary carcinoids?

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25203

    CTGF, HIF-1, IGF-1 and IGF-1R are expressed in the majority of tumors (A) . Their expression tended to be more frequent in TCs, although this was significant only for HIF-1 (B) .
    Figure Legend Snippet: CTGF, HIF-1, IGF-1 and IGF-1R are expressed in the majority of tumors (A) . Their expression tended to be more frequent in TCs, although this was significant only for HIF-1 (B) .

    Techniques Used: Expressing

    Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.
    Figure Legend Snippet: Immunostained sections of a typical ( A and B ) and an atypical ( C and D ) pulmonary carcinoid. (A) shows IGF-1, (B) IGF-1 receptor, (C) CTGF and (D) HIF-1 immunostained tumor sections. Virtually all tumor cells are immunoreactive and the immunoreactivity is cytoplasmic. The tumor capsule (A) and the fibrovascular stroma (B, C and D) are non-immunoreactive and can be used as internal control. Insets represent microphotographs of the neutralization tests (A, B and C) or replacement of the primary antibody by non-immune serum (D). Scale bars= 100 μm.

    Techniques Used: Neutralization

    21) Product Images from "Loss of Glutamatergic Pyramidal Neurons in Frontal and Temporal Cortex Resulting from Attenuation of FGFR1 Signaling Is Associated with Spontaneous Hyperactivity in Mice"

    Article Title: Loss of Glutamatergic Pyramidal Neurons in Frontal and Temporal Cortex Resulting from Attenuation of FGFR1 Signaling Is Associated with Spontaneous Hyperactivity in Mice

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5285-03.2004

    Radial glia morphology in the developing cerebral cortex at E18.5. A - C , Wild-type mice; D-F , tFgfr1 mutants. A, D , RC-2 immunoperoxidase staining of the medial prefrontal areas taken from midsagittal brain sections of E18.5 embryos. Note the elongated radial glial fibers reaching the pial layer in wild type ( A , arrowheads) and the fewer, fragmented, abnormal glia fibers in tFgfr1 ( D , arrowheads). Anterior is to the left. B - F , GLAST immunofluorescence staining of the temporal cortical areas taken from coronal brain sections at the level of lateral geniculate nuclei ( B,E ) or at more posterior levels ( C,F ). Note similar absence of elongated radial glial processes in tFgfr1 ( E,F ) as compared with wild type ( B,C ). Arrowheads point to radial glial processes in the CP. Medial is to the right and dorsal is to the top. Scale bars: A, C , 100 μm; B, D , 200 μm; C, F , 50 μm.
    Figure Legend Snippet: Radial glia morphology in the developing cerebral cortex at E18.5. A - C , Wild-type mice; D-F , tFgfr1 mutants. A, D , RC-2 immunoperoxidase staining of the medial prefrontal areas taken from midsagittal brain sections of E18.5 embryos. Note the elongated radial glial fibers reaching the pial layer in wild type ( A , arrowheads) and the fewer, fragmented, abnormal glia fibers in tFgfr1 ( D , arrowheads). Anterior is to the left. B - F , GLAST immunofluorescence staining of the temporal cortical areas taken from coronal brain sections at the level of lateral geniculate nuclei ( B,E ) or at more posterior levels ( C,F ). Note similar absence of elongated radial glial processes in tFgfr1 ( E,F ) as compared with wild type ( B,C ). Arrowheads point to radial glial processes in the CP. Medial is to the right and dorsal is to the top. Scale bars: A, C , 100 μm; B, D , 200 μm; C, F , 50 μm.

    Techniques Used: Mouse Assay, Immunoperoxidase Staining, Immunofluorescence, Staining

    22) 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

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

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

    Journal: FEBS Open Bio

    doi: 10.1016/j.fob.2011.10.003

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

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

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

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

    24) Product Images from "PRMT5 is essential for B cell development and germinal center dynamics"

    Article Title: PRMT5 is essential for B cell development and germinal center dynamics

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07884-6

    Prmt5 acts in the light zone to prevent B cell differentiation. a IHC images for Prmt5 and AID, as GC marker, on consecutive spleen sections from wt mice 14 days post-NP-CGG immunization. Representative of three mice from two experiments. Scale bar, 40 µm. b Representative IF on wt mouse spleen section 10 days after SRBC immunization, stained for GL7 CD35 (LZ marker) and Prmt5. Scale bar, 100 µm. c GSEA of transcriptional changes in Prmt5 F/F Cγ1-cre (F/F) versus Cγ1-cre (Ctrl) iGB cells against the indicated gene signatures. Enrichment was considered significant for P
    Figure Legend Snippet: Prmt5 acts in the light zone to prevent B cell differentiation. a IHC images for Prmt5 and AID, as GC marker, on consecutive spleen sections from wt mice 14 days post-NP-CGG immunization. Representative of three mice from two experiments. Scale bar, 40 µm. b Representative IF on wt mouse spleen section 10 days after SRBC immunization, stained for GL7 CD35 (LZ marker) and Prmt5. Scale bar, 100 µm. c GSEA of transcriptional changes in Prmt5 F/F Cγ1-cre (F/F) versus Cγ1-cre (Ctrl) iGB cells against the indicated gene signatures. Enrichment was considered significant for P

    Techniques Used: Cell Differentiation, Immunohistochemistry, Marker, Mouse Assay, Staining

    Prmt5 is necessary for GC expansion. a – l Cγ1-cre (Ctrl) versus Prmt5 F/F Cγ1-cre (F/F) mice. a Representative flow cytometry plots for GC B cells at various times post-SRBC immunization. Mean ± s.d. absolute GC B cell numbers from n mice from two to three experiments are plotted. b GCs per spleen section in individual mice (symbols), with means (bars). c Representative fluorescent microscopy images of spleen sections from mice in a stained for B cells (B220), T cells (CD3), FDCs (CD35), and activated B cells (GL7). Scale bar, 100 µm. d Representative IF confocal images in splenic sections from immunized mice, stained for the indicated markers, Prmt5 and sDMA (SYM11). GCs are contoured. Scale bars, 100 µm. e Representative histograms of activated pan-caspase staining in splenic GC B cells at day 8 post-SRBC immunization. Etoposide (3 µM) was used to induce apoptosis as a positive control. Mean + s.d. proportion of caspase + GC B cells for six mice per group from two independent experiments are plotted. f Representative histograms of Ki67 expression in splenic GC B cells. Mean + s.d. proportion of Ki67 low GC B cells for individual mice (symbols) from two experiments and means (bars) are plotted. g Representative flow cytometry plots of dark zone (DZ), light zone (LZ), and a CXCR4 − CD86 low GC B cells (gated on B220 + Gl7 + CD95 + ) in SRBC-immunized mice. The proportion of CXCR4 − CD86 low GC B cells for individual mice (symbols) from two to three experiments and means (bars) are plotted. h GC B cells in mice infected with H. polygyrus for 14 days analyzed as in g . Data from two experiments. i Representative histograms of Ki67 levels in GC B cell subsets in mice immunized with SRBC. j GC B cells and subpopulations as in g for Aicda − /− Cγ1-cre (Ctrl) or Aicda −/− Prmt5 F/F Cγ1-cre (F/F) mice immunized with SRBC. k Histograms of AID-GFP levels for individual Aicda -GFPtg Cγ1-cre (Ctrl) or Aicda -GFPtg Prmt5 F/F Cγ1-cre (F/F) mice at day 9 post-SRBC immunization. The AID dim /AID high ratio for individual mice (symbols) from two experiments and means (bars) are plotted. p -Values indicated throughout are by an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Prmt5 is necessary for GC expansion. a – l Cγ1-cre (Ctrl) versus Prmt5 F/F Cγ1-cre (F/F) mice. a Representative flow cytometry plots for GC B cells at various times post-SRBC immunization. Mean ± s.d. absolute GC B cell numbers from n mice from two to three experiments are plotted. b GCs per spleen section in individual mice (symbols), with means (bars). c Representative fluorescent microscopy images of spleen sections from mice in a stained for B cells (B220), T cells (CD3), FDCs (CD35), and activated B cells (GL7). Scale bar, 100 µm. d Representative IF confocal images in splenic sections from immunized mice, stained for the indicated markers, Prmt5 and sDMA (SYM11). GCs are contoured. Scale bars, 100 µm. e Representative histograms of activated pan-caspase staining in splenic GC B cells at day 8 post-SRBC immunization. Etoposide (3 µM) was used to induce apoptosis as a positive control. Mean + s.d. proportion of caspase + GC B cells for six mice per group from two independent experiments are plotted. f Representative histograms of Ki67 expression in splenic GC B cells. Mean + s.d. proportion of Ki67 low GC B cells for individual mice (symbols) from two experiments and means (bars) are plotted. g Representative flow cytometry plots of dark zone (DZ), light zone (LZ), and a CXCR4 − CD86 low GC B cells (gated on B220 + Gl7 + CD95 + ) in SRBC-immunized mice. The proportion of CXCR4 − CD86 low GC B cells for individual mice (symbols) from two to three experiments and means (bars) are plotted. h GC B cells in mice infected with H. polygyrus for 14 days analyzed as in g . Data from two experiments. i Representative histograms of Ki67 levels in GC B cell subsets in mice immunized with SRBC. j GC B cells and subpopulations as in g for Aicda − /− Cγ1-cre (Ctrl) or Aicda −/− Prmt5 F/F Cγ1-cre (F/F) mice immunized with SRBC. k Histograms of AID-GFP levels for individual Aicda -GFPtg Cγ1-cre (Ctrl) or Aicda -GFPtg Prmt5 F/F Cγ1-cre (F/F) mice at day 9 post-SRBC immunization. The AID dim /AID high ratio for individual mice (symbols) from two experiments and means (bars) are plotted. p -Values indicated throughout are by an unpaired, two-tailed Student's t test

    Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Microscopy, Staining, Positive Control, Expressing, Infection, Two Tailed Test

    Prmt5 loss induces p53-independant apoptosis. a WB probed for the indicated proteins on extracts from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) splenic B cells cultured with LPS and IL-4 for 3 days. b Gene transcript levels by RT-qPCR in splenic B cells from a . Mean + s.d. RNA level normalized to Actin for n mice are plotted relative to the Ctrl mean. c Representative histograms of Annexin-V levels in CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice in either Trp53 +/+ or Trp53 −/− background. Annexin-V + proportion for individual mice and mean + s.e.m cell concentration over time are plotted from one experiment. d Representative confocal microscopy of sDMA (SYM11) IF in iGB cells from wt mice treated with DMSO or 5 μM EPZ for 48 h. Large nuclei are from feeder cells. Scale bar, 10 µm. e Expansion of iGBs derived from wt, Trp53 −/− and Cdkn1a −/− splenic B cells, treated with DMSO or EPZ 24 h after plating. Means ± s.e.m cell counts of two mice from one experiment are plotted. f Sensitivity of iGB cells to EPZ 4 days after treating with EPZ doses. Relative mean ± s.e.m cell number and Cdkn1a expression by RT-qPCR are plotted for two mice from one experiment. g Expansion of iGBs from Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice and their Trp53 −/− counterparts. Mean ± s.e.m of cell count for n mice are plotted. h Cell cycle profile of Cγ1-cre Trp53 −/− (Ctrl) and Prmt5 F/F Cγ1-cre Trp53 −/− (F/F) iGBs pulsed with BrdU for 1 h at day 4 and stained with anti-BrdU and propidium iodide (PI). Means + s.d. for three mice per genotype from two independent experiments are plotted. i GC B cells in the spleen of Cγ1-cre Trp53 −/− (Ctrl) and Prmt5 F/F Cγ1-cre Trp53 −/− (F/F) mice 10 days after immunization with SRBC. Individual mice (symbols) and mean (bars) values are plotted from three experiments. j Representative flow cytometry and proportion of GC Cxcr4 − Cd86 low B cells in the mice in i . p -Values throughout are from an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Prmt5 loss induces p53-independant apoptosis. a WB probed for the indicated proteins on extracts from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) splenic B cells cultured with LPS and IL-4 for 3 days. b Gene transcript levels by RT-qPCR in splenic B cells from a . Mean + s.d. RNA level normalized to Actin for n mice are plotted relative to the Ctrl mean. c Representative histograms of Annexin-V levels in CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice in either Trp53 +/+ or Trp53 −/− background. Annexin-V + proportion for individual mice and mean + s.e.m cell concentration over time are plotted from one experiment. d Representative confocal microscopy of sDMA (SYM11) IF in iGB cells from wt mice treated with DMSO or 5 μM EPZ for 48 h. Large nuclei are from feeder cells. Scale bar, 10 µm. e Expansion of iGBs derived from wt, Trp53 −/− and Cdkn1a −/− splenic B cells, treated with DMSO or EPZ 24 h after plating. Means ± s.e.m cell counts of two mice from one experiment are plotted. f Sensitivity of iGB cells to EPZ 4 days after treating with EPZ doses. Relative mean ± s.e.m cell number and Cdkn1a expression by RT-qPCR are plotted for two mice from one experiment. g Expansion of iGBs from Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice and their Trp53 −/− counterparts. Mean ± s.e.m of cell count for n mice are plotted. h Cell cycle profile of Cγ1-cre Trp53 −/− (Ctrl) and Prmt5 F/F Cγ1-cre Trp53 −/− (F/F) iGBs pulsed with BrdU for 1 h at day 4 and stained with anti-BrdU and propidium iodide (PI). Means + s.d. for three mice per genotype from two independent experiments are plotted. i GC B cells in the spleen of Cγ1-cre Trp53 −/− (Ctrl) and Prmt5 F/F Cγ1-cre Trp53 −/− (F/F) mice 10 days after immunization with SRBC. Individual mice (symbols) and mean (bars) values are plotted from three experiments. j Representative flow cytometry and proportion of GC Cxcr4 − Cd86 low B cells in the mice in i . p -Values throughout are from an unpaired, two-tailed Student's t test

    Techniques Used: Western Blot, Cell Culture, Quantitative RT-PCR, Mouse Assay, Concentration Assay, Confocal Microscopy, Derivative Assay, Expressing, Cell Counting, Staining, Flow Cytometry, Cytometry, Two Tailed Test

    Prmt5 regulates gene expression and maintains splicing fidelity in B cells. a Volcano plot of gene expression changes in Prmt5 F/F Cγ1-cre versus Cγ1-cre iGB cells. The number of genes significantly changed by ≥2- or ≥4-fold (in red), are indicated. The violin plot shows the absolute change for the genes changing by ≥2-fold, p value from two-sided t -test. b GO terms significantly enriched for the 257 genes upregulated ≥2-fold in Prmt5-null iGBs, analyzed against a background list of all expressed genes in iGBs at the DAVID server. c GSEA of a curated set of 346 genes that are direct targets of p53 and are upregulated in response to p53. d Venn diagrams comparing genes differentially expressed by ≥1.5-fold to genes with at least one splicing alteration. Only significant events ( P -value
    Figure Legend Snippet: Prmt5 regulates gene expression and maintains splicing fidelity in B cells. a Volcano plot of gene expression changes in Prmt5 F/F Cγ1-cre versus Cγ1-cre iGB cells. The number of genes significantly changed by ≥2- or ≥4-fold (in red), are indicated. The violin plot shows the absolute change for the genes changing by ≥2-fold, p value from two-sided t -test. b GO terms significantly enriched for the 257 genes upregulated ≥2-fold in Prmt5-null iGBs, analyzed against a background list of all expressed genes in iGBs at the DAVID server. c GSEA of a curated set of 346 genes that are direct targets of p53 and are upregulated in response to p53. d Venn diagrams comparing genes differentially expressed by ≥1.5-fold to genes with at least one splicing alteration. Only significant events ( P -value

    Techniques Used: Expressing

    Prmt5 is required for B cell development. a BM lymphocytes and splenic B cells in MB1-cre (Ctrl) or Prmt5 F/F MB1-cre (F/F) mice. Representative flow cytometry plots and bar plots of mean + s.d. absolute lymphocyte counts for n mice from four experiments. b Representative flow cytometry plots gating for Hardy fractions A to E in BM from 3–4-months-old MB1-cre (Ctrl) and Prmt5 F/F MB1-cre (F/F) mice. Means + s.d. of absolute cell numbers for each fraction from n of mice from four independent experiments are plotted. c As in b , for MB1-cre B1-8 (ki+/+) and Prmt5 F/F MB1-cre B1-8 (ki F/F) mice, from three experiments. d As in b , for MB1-cre Trp53 −/− (pko+/+) and Prmt5 F/F MB1-cre Trp53 −/− (pko F/F) mice, from three experiments. e Proportion of large Pre-B cells in fractions C+C′ estimated by the pre-BCR expression in individual (symbols) MB1-cre Trp53 −/− (Ctrl) and Prmt5 F/F MB1-cre Trp53 −/− (F/F) mice. p -Values throughout are from an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Prmt5 is required for B cell development. a BM lymphocytes and splenic B cells in MB1-cre (Ctrl) or Prmt5 F/F MB1-cre (F/F) mice. Representative flow cytometry plots and bar plots of mean + s.d. absolute lymphocyte counts for n mice from four experiments. b Representative flow cytometry plots gating for Hardy fractions A to E in BM from 3–4-months-old MB1-cre (Ctrl) and Prmt5 F/F MB1-cre (F/F) mice. Means + s.d. of absolute cell numbers for each fraction from n of mice from four independent experiments are plotted. c As in b , for MB1-cre B1-8 (ki+/+) and Prmt5 F/F MB1-cre B1-8 (ki F/F) mice, from three experiments. d As in b , for MB1-cre Trp53 −/− (pko+/+) and Prmt5 F/F MB1-cre Trp53 −/− (pko F/F) mice, from three experiments. e Proportion of large Pre-B cells in fractions C+C′ estimated by the pre-BCR expression in individual (symbols) MB1-cre Trp53 −/− (Ctrl) and Prmt5 F/F MB1-cre Trp53 −/− (F/F) mice. p -Values throughout are from an unpaired, two-tailed Student's t test

    Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Expressing, Two Tailed Test

    Regulated Prmt5 expression in B cells. a Prmt5 transcript levels in B cell stages from the indicated datasets, each normalized to follicular (Fo) B cells. RT-qPCR data were normalized to Actin mRNA and obtained from a pool of two mice sorted for Hardy’s BM fractions (FrA to E), or from splenic B cells from two mice stimulated ex vivo with LPS and IL-4 for 48 h (Activated). CLP common lymphoid progenitor, T1–T3 transitional B cells, MZ marginal zone B cells, PC plasma cells, Sp spleen. b Prmt5 expression kinetics and sDMA-modified proteins by WB in extracts of WT splenic B cells stimulated with LPS (5 µg/mL) and IL-4 (5 ng/mL) probed with anti-PRMT5, -Actin (as loading control), and -sDMA (SYM11) antibodies. c Absolute number of B cell subpopulations in the spleen of CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice. Individual mice (dots) and mean (bars) values are plotted. Gatings in Supplementary Fig. 1B . d Representative WB of resting splenic B cell extracts from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice. Means + s.d. levels of Prmt5 normalized to Actin quantified from n mice by WB are plotted relative to the Ctrl. e Representative Prmt5, Actin, and sDMA (SYM11) WB and quantitation as in d , in extracts of splenic B cells activated with LPS (5 µg/mL) and IL-4 (5 ng/mL) for 72 h. f Prmt5 mRNA (by RT-qPCR) as a function of Prmt5 protein (by WB) at 72 h post-stimulation for individual mice. Spearman’s test correlation coefficient ( r ) and p -value ( p ) are indicated. g Representative immunohistochemical staining for PNA as GC marker and Prmt5 on consecutive mouse spleen sections at day 14 post-immunization. Scale bars, 500 µm
    Figure Legend Snippet: Regulated Prmt5 expression in B cells. a Prmt5 transcript levels in B cell stages from the indicated datasets, each normalized to follicular (Fo) B cells. RT-qPCR data were normalized to Actin mRNA and obtained from a pool of two mice sorted for Hardy’s BM fractions (FrA to E), or from splenic B cells from two mice stimulated ex vivo with LPS and IL-4 for 48 h (Activated). CLP common lymphoid progenitor, T1–T3 transitional B cells, MZ marginal zone B cells, PC plasma cells, Sp spleen. b Prmt5 expression kinetics and sDMA-modified proteins by WB in extracts of WT splenic B cells stimulated with LPS (5 µg/mL) and IL-4 (5 ng/mL) probed with anti-PRMT5, -Actin (as loading control), and -sDMA (SYM11) antibodies. c Absolute number of B cell subpopulations in the spleen of CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice. Individual mice (dots) and mean (bars) values are plotted. Gatings in Supplementary Fig. 1B . d Representative WB of resting splenic B cell extracts from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice. Means + s.d. levels of Prmt5 normalized to Actin quantified from n mice by WB are plotted relative to the Ctrl. e Representative Prmt5, Actin, and sDMA (SYM11) WB and quantitation as in d , in extracts of splenic B cells activated with LPS (5 µg/mL) and IL-4 (5 ng/mL) for 72 h. f Prmt5 mRNA (by RT-qPCR) as a function of Prmt5 protein (by WB) at 72 h post-stimulation for individual mice. Spearman’s test correlation coefficient ( r ) and p -value ( p ) are indicated. g Representative immunohistochemical staining for PNA as GC marker and Prmt5 on consecutive mouse spleen sections at day 14 post-immunization. Scale bars, 500 µm

    Techniques Used: Expressing, Quantitative RT-PCR, Mouse Assay, Ex Vivo, Modification, Western Blot, Quantitation Assay, Immunohistochemistry, Staining, Marker

    Prmt5 is required for the survival of activated B cells in vivo. All panels compare Prmt5 F/F CD19-cre (F/F) versus CD19-cre (Ctrl) mice. a Representative flow cytometry plots of splenic GC B cells (B220 + , GL7 high , CD95 + ) 14 days after NP-CGG immunization. Total number of GC B cell number for individual mice (symbols) and mean values (bars) from three experiments are plotted. b Representative immunofluorescence microscopy of mouse spleen sections stained for Prmt5, B220, GL7, and IgD from unimmunized (two per genotype from one experiment) or immunized with NP-CGG, day 14 (four per genotype from two experiments). Scale bars, 100 μm. The Prmt5 signal per follicular (Fo) and GC B cell was quantified in individual follicles from four immunized mice of each genotype. The lefthand plot shows mean normalized Prmt5 signal in Fo and GC B cells from 4–7 follicles per mouse (symbols), with lines joining mean values (bars) of the two B cell types in individual mice. The righthand plot shows the mean GC/Fo signal ratio per mouse. c Absolute number of splenic lymphocytes for individual mice unimmunized or 14 days post-immunization (symbols) with means (bars) from two experiments. d Representative histograms of GL7 expression in non-GC B cells (B220 + CD95 − ) from mice injected or not with Alum (day 14). One experiment, two mice per genotype per treatment. e Absolute number of B and T cells in mesenteric lymph node (MLN) and spleen of individual mice (symbols) 14 days post-infection with H. polygyrus , with medians (bars), from two experiments. f Sequential gating to analyze T, GC, and non-GC B cells, as well as the GL7− and GL7+ fractions of the latter (top) and representative histograms of pan-caspase staining in MLN of the mice analyzed in e . Gates were set using cells treated with etoposide (3 µM). Means + s.d. proportion of pan-caspase + cells from six mice from two experiments are plotted, normalized to the control’s average for each subset. p -Values are by Mann–Whitney test ( b ) or an unpaired, two-tailed Student's t test ( c , e , f ); ns, not significant
    Figure Legend Snippet: Prmt5 is required for the survival of activated B cells in vivo. All panels compare Prmt5 F/F CD19-cre (F/F) versus CD19-cre (Ctrl) mice. a Representative flow cytometry plots of splenic GC B cells (B220 + , GL7 high , CD95 + ) 14 days after NP-CGG immunization. Total number of GC B cell number for individual mice (symbols) and mean values (bars) from three experiments are plotted. b Representative immunofluorescence microscopy of mouse spleen sections stained for Prmt5, B220, GL7, and IgD from unimmunized (two per genotype from one experiment) or immunized with NP-CGG, day 14 (four per genotype from two experiments). Scale bars, 100 μm. The Prmt5 signal per follicular (Fo) and GC B cell was quantified in individual follicles from four immunized mice of each genotype. The lefthand plot shows mean normalized Prmt5 signal in Fo and GC B cells from 4–7 follicles per mouse (symbols), with lines joining mean values (bars) of the two B cell types in individual mice. The righthand plot shows the mean GC/Fo signal ratio per mouse. c Absolute number of splenic lymphocytes for individual mice unimmunized or 14 days post-immunization (symbols) with means (bars) from two experiments. d Representative histograms of GL7 expression in non-GC B cells (B220 + CD95 − ) from mice injected or not with Alum (day 14). One experiment, two mice per genotype per treatment. e Absolute number of B and T cells in mesenteric lymph node (MLN) and spleen of individual mice (symbols) 14 days post-infection with H. polygyrus , with medians (bars), from two experiments. f Sequential gating to analyze T, GC, and non-GC B cells, as well as the GL7− and GL7+ fractions of the latter (top) and representative histograms of pan-caspase staining in MLN of the mice analyzed in e . Gates were set using cells treated with etoposide (3 µM). Means + s.d. proportion of pan-caspase + cells from six mice from two experiments are plotted, normalized to the control’s average for each subset. p -Values are by Mann–Whitney test ( b ) or an unpaired, two-tailed Student's t test ( c , e , f ); ns, not significant

    Techniques Used: In Vivo, Mouse Assay, Flow Cytometry, Cytometry, Immunofluorescence, Microscopy, Staining, Expressing, Injection, Infection, MANN-WHITNEY, Two Tailed Test

    Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Flow Cytometry, Cytometry, Infection, Staining, Two Tailed Test

    Prmt5 protects B cells from apoptosis and promotes proliferation. a – d One million naïve splenic B cells from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice were plated with LPS (5 µg/mL) + IL-4 (5 ng/mL). a Cell expansion over time monitored by cell counting (left) and number of cell divisions by day 3 monitored by CFSE stain dilution (right) from two experiments, four mice per genotype. b Cell cycle profile of cells pulsed with BrdU at day 3 before staining with anti-BrdU and propidium iodide (PI). Data for three mice (symbols) per genotype and means (bars) from one experiment are plotted. c Representative Annexin-V + staining histograms of B cells 3 days post-plating. The proportion of Annexin-V + cells for individual mice (symbols) and means (bars) from five experiments are plotted normalized to the Ctrl mean. d Proportion of Annexin-V + B cells as a function of Prmt5 protein levels measured by WB and normalized to Actin 72 h post-stimulation. Spearman’s correlation coefficient ( r ) and p -value ( p ) are indicated. e WB of Prmt5 and Revert protein staining as loading control in extracts of splenic B cells from Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice, stimulated as in a for 72 h. The normalized proportion of Annexin-V + cells for individual mice (symbols) from three experiments and mean values (bars) are plotted. f Top, resting splenic B cells from Prmt5 F/+ CD19-cre (Ctrl) or Prmt5 F/F CD19-cre (F/+) and Prmt5 F/F CD19-cre (F/F) mice plated onto 40LB cells with 1 ng/mL IL-4 to generate GC-like B cells (iGBs). Mean ± s.e.m. cell counts per day are plotted for two experiments with two mice each. Bottom, proportion of Annexin-V + iGBs for individual mice (symbols) and means (bars) for both experiments are plotted, pooling +/+ and F/+ as controls. g Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice iGBs analyzed as in f . h Representative cell cycle profile as in b in Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) iGBs at day 4. Means + s.d. of six mice per genotype from three experiments are plotted. Unpaired, two-tailed Student's t test was performed in a , c , e – g , only significant p -values are shown
    Figure Legend Snippet: Prmt5 protects B cells from apoptosis and promotes proliferation. a – d One million naïve splenic B cells from CD19-cre (Ctrl) and Prmt5 F/F CD19-cre (F/F) mice were plated with LPS (5 µg/mL) + IL-4 (5 ng/mL). a Cell expansion over time monitored by cell counting (left) and number of cell divisions by day 3 monitored by CFSE stain dilution (right) from two experiments, four mice per genotype. b Cell cycle profile of cells pulsed with BrdU at day 3 before staining with anti-BrdU and propidium iodide (PI). Data for three mice (symbols) per genotype and means (bars) from one experiment are plotted. c Representative Annexin-V + staining histograms of B cells 3 days post-plating. The proportion of Annexin-V + cells for individual mice (symbols) and means (bars) from five experiments are plotted normalized to the Ctrl mean. d Proportion of Annexin-V + B cells as a function of Prmt5 protein levels measured by WB and normalized to Actin 72 h post-stimulation. Spearman’s correlation coefficient ( r ) and p -value ( p ) are indicated. e WB of Prmt5 and Revert protein staining as loading control in extracts of splenic B cells from Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice, stimulated as in a for 72 h. The normalized proportion of Annexin-V + cells for individual mice (symbols) from three experiments and mean values (bars) are plotted. f Top, resting splenic B cells from Prmt5 F/+ CD19-cre (Ctrl) or Prmt5 F/F CD19-cre (F/+) and Prmt5 F/F CD19-cre (F/F) mice plated onto 40LB cells with 1 ng/mL IL-4 to generate GC-like B cells (iGBs). Mean ± s.e.m. cell counts per day are plotted for two experiments with two mice each. Bottom, proportion of Annexin-V + iGBs for individual mice (symbols) and means (bars) for both experiments are plotted, pooling +/+ and F/+ as controls. g Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice iGBs analyzed as in f . h Representative cell cycle profile as in b in Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) iGBs at day 4. Means + s.d. of six mice per genotype from three experiments are plotted. Unpaired, two-tailed Student's t test was performed in a , c , e – g , only significant p -values are shown

    Techniques Used: Mouse Assay, Cell Counting, Staining, Western Blot, Two Tailed Test

    25) Product Images from "Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice"

    Article Title: Consequences of early postnatal lipopolysaccharide exposure on developing lungs in mice

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00560.2017

    Deficits in proliferation and survival of the cells of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were harvested on PND14 for lung immunohistochemistry studies. Representative lung sections showing Ki67-stained cells from mice treated on PND3 with PBS ( A ) and L10 ( B ) and on PND7 with PBS ( C ) and L10 ( D ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated on PND3 with PBS ( E ) and L10 ( F ) and on PND7 with PBS ( G ) and L10 ( H ). Quantification of Ki67-positive (percentage) ( I ) and cleaved caspase 3-positive (number per high power field) ( J ) lung cells. Values are presented as means ± SD ( n = 6/group). Significant differences between age-matched PBS- and L10-exposed animals are indicated by *** P
    Figure Legend Snippet: Deficits in proliferation and survival of the cells of saccular and alveolar lungs exposed to a single dose of LPS. Newborn mice were injected intraperitoneally with a single dose of vehicle control (PBS) or 10 mg/kg LPS (L10) on postnatal day (PND)3 or PND7, and the lung tissues were harvested on PND14 for lung immunohistochemistry studies. Representative lung sections showing Ki67-stained cells from mice treated on PND3 with PBS ( A ) and L10 ( B ) and on PND7 with PBS ( C ) and L10 ( D ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated on PND3 with PBS ( E ) and L10 ( F ) and on PND7 with PBS ( G ) and L10 ( H ). Quantification of Ki67-positive (percentage) ( I ) and cleaved caspase 3-positive (number per high power field) ( J ) lung cells. Values are presented as means ± SD ( n = 6/group). Significant differences between age-matched PBS- and L10-exposed animals are indicated by *** P

    Techniques Used: Mouse Assay, Injection, Immunohistochemistry, Staining

    Effect of chronic LPS exposure on lung cell proliferation and apoptosis. Newborn mice were treated intraperitoneally with 10 mg/kg of LPS (L10) or a vehicle control (PBS) on postnatal days (PNDs) 3–5 , and the lung tissues were harvested for immunohistochemistry studies on PND7. Representative lung sections showing Ki67-stained cells from mice treated with PBS ( A ) and L10 ( B ). Quantification of the percentage of Ki67-positive lung cells ( C ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated with PBS ( D ) and L10 ( E ). Quantification of cleaved caspase 3-positive lung cells ( F ). HPF, high-power field. Values are presented as means ± SD ( n = 6/group). Significant differences between PBS- and L10-treated animals are indicated by *** P
    Figure Legend Snippet: Effect of chronic LPS exposure on lung cell proliferation and apoptosis. Newborn mice were treated intraperitoneally with 10 mg/kg of LPS (L10) or a vehicle control (PBS) on postnatal days (PNDs) 3–5 , and the lung tissues were harvested for immunohistochemistry studies on PND7. Representative lung sections showing Ki67-stained cells from mice treated with PBS ( A ) and L10 ( B ). Quantification of the percentage of Ki67-positive lung cells ( C ). Representative lung sections showing cleaved caspase 3-positive cells from mice treated with PBS ( D ) and L10 ( E ). Quantification of cleaved caspase 3-positive lung cells ( F ). HPF, high-power field. Values are presented as means ± SD ( n = 6/group). Significant differences between PBS- and L10-treated animals are indicated by *** P

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining

    26) Product Images from "A Small Molecule Inhibitor of the β-Catenin-TCF4 Interaction Suppresses Colorectal Cancer Growth In Vitro and In Vivo"

    Article Title: A Small Molecule Inhibitor of the β-Catenin-TCF4 Interaction Suppresses Colorectal Cancer Growth In Vitro and In Vivo

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2017.09.029

    HI-B1 inhibits β-catenin-driven tumorigenesis in vivo . (a) β-Catenin expression levels in colon cancer PDX samples were examined by immunohistochemistry (IHC) and its integrated optical density (IOD) was quantified using Image-Pro Plus software (v.6.1) program (Media Cybernetics, Bethesda, MD). Scale bar: 50 μm. (b and c) HI-B1 was administered per os (p.o) (orally) three times a week at a dose of 50 mg/kg ( n = 8/group) to JG5 (b) and JG14 (c) tumors. Tumor size was measured three times a week and the weight was measured at the end of experiments. (One-way ANOVA and Dunnett's test; *P
    Figure Legend Snippet: HI-B1 inhibits β-catenin-driven tumorigenesis in vivo . (a) β-Catenin expression levels in colon cancer PDX samples were examined by immunohistochemistry (IHC) and its integrated optical density (IOD) was quantified using Image-Pro Plus software (v.6.1) program (Media Cybernetics, Bethesda, MD). Scale bar: 50 μm. (b and c) HI-B1 was administered per os (p.o) (orally) three times a week at a dose of 50 mg/kg ( n = 8/group) to JG5 (b) and JG14 (c) tumors. Tumor size was measured three times a week and the weight was measured at the end of experiments. (One-way ANOVA and Dunnett's test; *P

    Techniques Used: In Vivo, Expressing, Immunohistochemistry, Software

    27) Product Images from "PRMT5 is essential for B cell development and germinal center dynamics"

    Article Title: PRMT5 is essential for B cell development and germinal center dynamics

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07884-6

    Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Flow Cytometry, Cytometry, Infection, Staining, Two Tailed Test

    28) Product Images from "α-catenin acts as a tumor suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signaling"

    Article Title: α-catenin acts as a tumor suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signaling

    Journal: Nature cell biology

    doi: 10.1038/ncb2909

    Loss of α-catenin promotes tumor growth in basal-like breast cancer cells by activating NF-κB signaling ( a ) Immunoblotting of α-catenin, RelA and HSP90 in MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. ( b ) Soft agar colony formation by MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. n = 4 wells per group. ( c ) Tumor growth by subcutaneously implanted MDA-MB-231 (3 × 10 6 cells injected) or BT549 (4 × 10 6 cells injected) cells infected with α-catenin shRNA alone or in combination with RelA shRNA. P values correspond to comparisons between α-catenin shRNA alone and α-catenin shRNA in combination with RelA shRNA. ( d, e ) Tumor weight ( d ) and tumor images ( e ) 5 weeks after mice were injected subcutaneously with MDA-MB-231 or BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. Scale bar: 1 cm. n = 5 (for MDA-MB-231 cells) or 8 (for BT549 cells) mice per group in ( c ) and ( d ). ( f, g ) TNFα ( f ) and human-specific IκBα ( g ) immunohistochemical staining of subcutaneous tumors formed by MDA-MB-231 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA, at 5 weeks after implantation. Scale bar: 50 μm. Data in ( b ) – ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: Loss of α-catenin promotes tumor growth in basal-like breast cancer cells by activating NF-κB signaling ( a ) Immunoblotting of α-catenin, RelA and HSP90 in MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. ( b ) Soft agar colony formation by MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. n = 4 wells per group. ( c ) Tumor growth by subcutaneously implanted MDA-MB-231 (3 × 10 6 cells injected) or BT549 (4 × 10 6 cells injected) cells infected with α-catenin shRNA alone or in combination with RelA shRNA. P values correspond to comparisons between α-catenin shRNA alone and α-catenin shRNA in combination with RelA shRNA. ( d, e ) Tumor weight ( d ) and tumor images ( e ) 5 weeks after mice were injected subcutaneously with MDA-MB-231 or BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. Scale bar: 1 cm. n = 5 (for MDA-MB-231 cells) or 8 (for BT549 cells) mice per group in ( c ) and ( d ). ( f, g ) TNFα ( f ) and human-specific IκBα ( g ) immunohistochemical staining of subcutaneous tumors formed by MDA-MB-231 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA, at 5 weeks after implantation. Scale bar: 50 μm. Data in ( b ) – ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Multiple Displacement Amplification, Transduction, shRNA, Injection, Infection, Mouse Assay, Immunohistochemistry, Staining, Two Tailed Test

    α-catenin inhibits NF-κB signaling in basal-like breast cancer cells ( a ) qPCR of NF-κB response genes in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( b ) ELISA of TNFα and IL-8 secreted by α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 wells per group. Data in ( a ) and ( b ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 .
    Figure Legend Snippet: α-catenin inhibits NF-κB signaling in basal-like breast cancer cells ( a ) qPCR of NF-κB response genes in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( b ) ELISA of TNFα and IL-8 secreted by α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 wells per group. Data in ( a ) and ( b ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 .

    Techniques Used: Real-time Polymerase Chain Reaction, Multiple Displacement Amplification, Enzyme-linked Immunosorbent Assay, Two Tailed Test

    α-catenin stabilizes IκBα protein by inhibiting IκBα ubiquitination and abrogating IκBα interaction with the proteasome ( a ) Upper panel: luciferase assays of NF-κB activity in MDA-MB-157 cells transfected with a pNFκB luciferase reporter alone or in combination with FLAG-α-catenin, with or without TNFα treatment. Lower panel: immunoblotting of FLAG, p-IκBα, IκBα, p-RelA, RelA and HSP90. n = 3 wells per group. ( b ) Immunoblotting of α-catenin, IκBα and HSP90 in α-catenin shRNA-transduced BT549 cells, with or without TNFα treatment. ( c ) Immunoblotting of IκBα and HSP90 in α-catenin-transduced MDA-MB-231 and MDA-MB-157 cells. ( d ) qPCR of CTNNA1 and NFKBIA in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( e ) α-catenin-transduced MDA-MB-157 cells were treated with 100 μg/ml of cycloheximide (CHX), harvested at different time points and immunoblotted with antibodies to α-catenin, IκBα and HSP90. ( f ) Quantification of IκBα protein levels in ( e ). ( g ) α-catenin was immunoprecipitated from α-catenin-transduced MDA-MB-157 cells and immunoblotted with antibodies to α-catenin and IκBα. ( h ) α-catenin was immunoprecipitated from MDA-MB-231 and BT549 cells and immunoblotted with antibodies to α-catenin, β-catenin, IκBα and RelA. ( i ) α-catenin was immunoprecipitated from untreated or TNFα-treated BT549 cells and immunoblotted with antibodies to α-catenin and IκBα. ( j ) α-catenin was immunoprecipitated from T47D or MCF10A cells and immunoblotted with antibodies to α-catenin, IκBα and RelA. ( k ) Upper panel: schematic representation of three vinculin domains of α-catenin. Lower panel: 293T cells were cotransfected with MYC-IκBα and SFB-tagged full-length α-catenin or fragment VH1, VH2 or VH3. α-catenin and the three fragments were purified with S-protein beads and immunoblotted with antibodies to FLAG and MYC. ( l ) HA-tagged wild-type ubiquitin (Ub) or the K48R or K63R mutant was cotransfected with MYC-α-catenin and SFB-IκBα into 293T cells. Cells were treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to HA and FLAG. ( m ) 293T cells were cotransfected with MYC-α-catenin and SFB-IκBα and then treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to the 20S proteasome subunit α4, MYC and FLAG. Data in ( a ) and ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: α-catenin stabilizes IκBα protein by inhibiting IκBα ubiquitination and abrogating IκBα interaction with the proteasome ( a ) Upper panel: luciferase assays of NF-κB activity in MDA-MB-157 cells transfected with a pNFκB luciferase reporter alone or in combination with FLAG-α-catenin, with or without TNFα treatment. Lower panel: immunoblotting of FLAG, p-IκBα, IκBα, p-RelA, RelA and HSP90. n = 3 wells per group. ( b ) Immunoblotting of α-catenin, IκBα and HSP90 in α-catenin shRNA-transduced BT549 cells, with or without TNFα treatment. ( c ) Immunoblotting of IκBα and HSP90 in α-catenin-transduced MDA-MB-231 and MDA-MB-157 cells. ( d ) qPCR of CTNNA1 and NFKBIA in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( e ) α-catenin-transduced MDA-MB-157 cells were treated with 100 μg/ml of cycloheximide (CHX), harvested at different time points and immunoblotted with antibodies to α-catenin, IκBα and HSP90. ( f ) Quantification of IκBα protein levels in ( e ). ( g ) α-catenin was immunoprecipitated from α-catenin-transduced MDA-MB-157 cells and immunoblotted with antibodies to α-catenin and IκBα. ( h ) α-catenin was immunoprecipitated from MDA-MB-231 and BT549 cells and immunoblotted with antibodies to α-catenin, β-catenin, IκBα and RelA. ( i ) α-catenin was immunoprecipitated from untreated or TNFα-treated BT549 cells and immunoblotted with antibodies to α-catenin and IκBα. ( j ) α-catenin was immunoprecipitated from T47D or MCF10A cells and immunoblotted with antibodies to α-catenin, IκBα and RelA. ( k ) Upper panel: schematic representation of three vinculin domains of α-catenin. Lower panel: 293T cells were cotransfected with MYC-IκBα and SFB-tagged full-length α-catenin or fragment VH1, VH2 or VH3. α-catenin and the three fragments were purified with S-protein beads and immunoblotted with antibodies to FLAG and MYC. ( l ) HA-tagged wild-type ubiquitin (Ub) or the K48R or K63R mutant was cotransfected with MYC-α-catenin and SFB-IκBα into 293T cells. Cells were treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to HA and FLAG. ( m ) 293T cells were cotransfected with MYC-α-catenin and SFB-IκBα and then treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to the 20S proteasome subunit α4, MYC and FLAG. Data in ( a ) and ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Luciferase, Activity Assay, Multiple Displacement Amplification, Transfection, shRNA, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Mutagenesis, Two Tailed Test

    α-catenin inhibits RelA-p50 nuclear localization and downregulates RelB ( a ) Immunoblotting of α-catenin and RelA in cytoplasmic and nuclear fractions of BT549 cells transduced with two independent α-catenin shRNAs, with or without TNFα treatment. ( b ) Immunoblotting of α-catenin, p105, p50, RelA, RelB and IκBα in cytoplasmic and nuclear fractions of α-catenin-transduced MDA-MB-157 cells. α-tubulin and Lamin A were used as cytoplasmic and nuclear markers, respectively, in ( a ) and ( b ). ( c ) Schematic representation of the RELB promoter containing two RelA binding sites (red rectangles). The two boxed arrows indicate the primers used for ChIP-qPCR. Hs: Homo sapiens , Mm: Mus musculus . ( d ) ChIP-qPCR analysis of RelA binding to the RELB promoter in α-catenin-transduced MDA-MB-157 cells. qPCR was performed with primers specific to the RelA binding motifs. Data were normalized to the input. n = 3 samples per group. ( e ) qPCR of RELB in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( f ) Immunoblotting of RelB and HSP90 in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. SE: short exposure; LE: long exposure. Data in ( d ) and ( e ) are the mean of biological replicates from a representative experiment,, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: α-catenin inhibits RelA-p50 nuclear localization and downregulates RelB ( a ) Immunoblotting of α-catenin and RelA in cytoplasmic and nuclear fractions of BT549 cells transduced with two independent α-catenin shRNAs, with or without TNFα treatment. ( b ) Immunoblotting of α-catenin, p105, p50, RelA, RelB and IκBα in cytoplasmic and nuclear fractions of α-catenin-transduced MDA-MB-157 cells. α-tubulin and Lamin A were used as cytoplasmic and nuclear markers, respectively, in ( a ) and ( b ). ( c ) Schematic representation of the RELB promoter containing two RelA binding sites (red rectangles). The two boxed arrows indicate the primers used for ChIP-qPCR. Hs: Homo sapiens , Mm: Mus musculus . ( d ) ChIP-qPCR analysis of RelA binding to the RELB promoter in α-catenin-transduced MDA-MB-157 cells. qPCR was performed with primers specific to the RelA binding motifs. Data were normalized to the input. n = 3 samples per group. ( e ) qPCR of RELB in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( f ) Immunoblotting of RelB and HSP90 in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. SE: short exposure; LE: long exposure. Data in ( d ) and ( e ) are the mean of biological replicates from a representative experiment,, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Transduction, Multiple Displacement Amplification, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Two Tailed Test

    29) Product Images from "PRMT5 is essential for B cell development and germinal center dynamics"

    Article Title: PRMT5 is essential for B cell development and germinal center dynamics

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07884-6

    Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test
    Figure Legend Snippet: Antibody response and GC defects caused by Prmt5 deficiency. a – i Cγ1-cre (Ctrl) and Prmt5 F/F Cγ1-cre (F/F) mice were used throughout. a Total anti-NP IgG1 in the serum of mice, measured by ELISA 14 days after NP-CGG immunization. Mean ± s.d. OD values for serial dilutions are plotted for n mice from three experiments. b Representative pictures of ELISPOT for NP-specific IgG1 antibody secreting cells (ASC) at day 14 post-immunization. The number of ASC of individual mice (symbols) and means (bars) are plotted. c Anti-NP IgG1 in the serum of mice at various times post-immunization. Mean ± s.d. values for n mice at each time point from two experiments are plotted. d Total levels of antibody isotypes in the serum of n non-immunized mice. e Mean + s.d. number of lymphocytes per spleen at day 14 post-immunization, enumerated by flow cytometry for n mice from two experiments. f Representative flow cytometry plots (gated on B220 + ) of splenic GC B cell proportions at 14 days post-immunization with NP-CGG. The number of GC B cells per spleen for individual mice (symbol) and medians (bars) from three experiments are plotted. g As in f , for MLN of mice infected with H. polygyrus for 14 days. Data from two experiments are plotted. h Mean + s.d. number of lymphocytes per MLN in the mice from g . i Representative IF in MLN from mice infected with H. polygyrus from g , stained for the indicated antigens. Scale bar, 100 µm. GC numbers per MLN scored in individual mice (symbols) are plotted to the right. p -Values throughout are by an unpaired, two-tailed Student's t test

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Flow Cytometry, Cytometry, Infection, Staining, Two Tailed Test

    30) Product Images from "Productive Propagation of Rift Valley Fever Phlebovirus Vaccine Strain MP-12 in Rousettus aegyptiacus Fruit Bats"

    Article Title: Productive Propagation of Rift Valley Fever Phlebovirus Vaccine Strain MP-12 in Rousettus aegyptiacus Fruit Bats

    Journal: Viruses

    doi: 10.3390/v10120681

    Histopathological findings in the liver of a Rousettus aegyptiacus fruit bat immunized with the MP-12 vaccine strain at day seven post immunization. ( A ) Histopathology shows few, randomly distributed foci of hepatocellular necrosis and loss with macrophage and lymphocyte infiltration (arrow). Furthermore, the hepatocytes display moderate, coalescing to diffuse, floccular cytoplasmic vacuolization, interpreted as a species-specific, relatively high level of glycogen storage. Hematoxylin-eosin. Bar = 100 μm; ( B ) Immunohistochemistry for Rift Valley fever phlebovirus (RVFV) Gc antigen reveals minor amounts of intra- and extracellular, strongly immunoreactive granula within the lesions (arrow), interpreted as debris remaining after virus-induced hepatocellular death. Immunohistochemistry, monoclonal mouse anti-RVFV Gc-protein antibody, avidin-biotin-peroxidase-complex method, 3-amino-9-ethyl-carbazol chromogen (red), hematoxylin counterstain (blue). Bar = 20 μm.
    Figure Legend Snippet: Histopathological findings in the liver of a Rousettus aegyptiacus fruit bat immunized with the MP-12 vaccine strain at day seven post immunization. ( A ) Histopathology shows few, randomly distributed foci of hepatocellular necrosis and loss with macrophage and lymphocyte infiltration (arrow). Furthermore, the hepatocytes display moderate, coalescing to diffuse, floccular cytoplasmic vacuolization, interpreted as a species-specific, relatively high level of glycogen storage. Hematoxylin-eosin. Bar = 100 μm; ( B ) Immunohistochemistry for Rift Valley fever phlebovirus (RVFV) Gc antigen reveals minor amounts of intra- and extracellular, strongly immunoreactive granula within the lesions (arrow), interpreted as debris remaining after virus-induced hepatocellular death. Immunohistochemistry, monoclonal mouse anti-RVFV Gc-protein antibody, avidin-biotin-peroxidase-complex method, 3-amino-9-ethyl-carbazol chromogen (red), hematoxylin counterstain (blue). Bar = 20 μm.

    Techniques Used: Histopathology, Immunohistochemistry, Avidin-Biotin Assay

    31) Product Images from "Interferon-stimulated genes—essential antiviral effectors implicated in resistance to Theiler’s virus-induced demyelinating disease"

    Article Title: Interferon-stimulated genes—essential antiviral effectors implicated in resistance to Theiler’s virus-induced demyelinating disease

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-015-0462-x

    ISG15 and PKR protein expression in the spinal cord of mock- and TMEV-infected C57BL/6 mice. a Few cells express ISG15 proteins in TMEV-infected SJL/J mice at 98 dpi. b Many endothelial cells, astrocytes, and neurons express ISG15 proteins in TMEV-infected C57BL/6 mice at 98 dpi. c Low expression of PKR proteins in TMEV-infected SJL/J mice at 98 dpi. d High expression of PKR proteins in oligodendrocytes, microglia/macrophages, and neurons of TMEV-infected C57BL/6 mice at 98 dpi. Immunohistochemistry visualized by the avidin-biotin-peroxidase complex method with 3,3-diaminobenzidine as substrate and counterstaining with Mayer’s hematoxylin. Bar 100 μm
    Figure Legend Snippet: ISG15 and PKR protein expression in the spinal cord of mock- and TMEV-infected C57BL/6 mice. a Few cells express ISG15 proteins in TMEV-infected SJL/J mice at 98 dpi. b Many endothelial cells, astrocytes, and neurons express ISG15 proteins in TMEV-infected C57BL/6 mice at 98 dpi. c Low expression of PKR proteins in TMEV-infected SJL/J mice at 98 dpi. d High expression of PKR proteins in oligodendrocytes, microglia/macrophages, and neurons of TMEV-infected C57BL/6 mice at 98 dpi. Immunohistochemistry visualized by the avidin-biotin-peroxidase complex method with 3,3-diaminobenzidine as substrate and counterstaining with Mayer’s hematoxylin. Bar 100 μm

    Techniques Used: Expressing, Infection, Mouse Assay, Immunohistochemistry, Avidin-Biotin Assay

    ISG15, PKR, and OAS1 protein expression in the spinal cord of mock- and TMEV-infected SJL/J mice. a Low expression of ISG15 proteins in mock-infected mice at 42 dpi. b High expression in endothelial cells and astrocytes of TMEV-infected SJL/J mice at 42 dpi. c Few cells express PKR proteins in mock-infected SJL/J mice at 196 dpi. d Many oligodendrocytes, microglia/macrophages, and neurons express PKR proteins in TMEV-infected SJL/J mice at 196 dpi. e Few intralesional cells express ISG15 at 98 dpi. f High PKR expression in intralesional gitter cells and some perivascular immune cells in TMEV-infected SJL/J mice at 98 dpi. g Very low expression of OAS1 proteins of mock-infected SJL/J mice at 98 dpi. h Few neurons, microglia, and perivascular immune cells express OAS1 proteins in TMEV-infected SJL/J mice at 98 dpi. Immunohistochemistry visualized by the avidin-biotin-peroxidase complex method with 3,3-diaminobenzidine as substrate and counterstaining with Mayer’s hematoxylin. Bar 100 μm
    Figure Legend Snippet: ISG15, PKR, and OAS1 protein expression in the spinal cord of mock- and TMEV-infected SJL/J mice. a Low expression of ISG15 proteins in mock-infected mice at 42 dpi. b High expression in endothelial cells and astrocytes of TMEV-infected SJL/J mice at 42 dpi. c Few cells express PKR proteins in mock-infected SJL/J mice at 196 dpi. d Many oligodendrocytes, microglia/macrophages, and neurons express PKR proteins in TMEV-infected SJL/J mice at 196 dpi. e Few intralesional cells express ISG15 at 98 dpi. f High PKR expression in intralesional gitter cells and some perivascular immune cells in TMEV-infected SJL/J mice at 98 dpi. g Very low expression of OAS1 proteins of mock-infected SJL/J mice at 98 dpi. h Few neurons, microglia, and perivascular immune cells express OAS1 proteins in TMEV-infected SJL/J mice at 98 dpi. Immunohistochemistry visualized by the avidin-biotin-peroxidase complex method with 3,3-diaminobenzidine as substrate and counterstaining with Mayer’s hematoxylin. Bar 100 μm

    Techniques Used: Expressing, Infection, Mouse Assay, Immunohistochemistry, Avidin-Biotin Assay

    32) Product Images from "Modulation of lethal HPAIV H5N8 clade 2.3.4.4B infection in AIV pre-exposed mallards"

    Article Title: Modulation of lethal HPAIV H5N8 clade 2.3.4.4B infection in AIV pre-exposed mallards

    Journal: Emerging Microbes & Infections

    doi: 10.1080/22221751.2020.1713706

    Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.
    Figure Legend Snippet: Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.

    Techniques Used: Infection, Lysis, Immunohistochemistry, Avidin-Biotin Assay

    33) Product Images from "Wnt/?-Catenin Signaling Regulates Yes-associated Protein (YAP) Gene Expression in Colorectal Carcinoma Cells *"

    Article Title: Wnt/?-Catenin Signaling Regulates Yes-associated Protein (YAP) Gene Expression in Colorectal Carcinoma Cells *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.327767

    β-Catenin, TCF4, and RNAP bind the first intron of the YAP gene in vivo . A , diagram of the YAP gene locus and the β-catenin-enriched regions identified in a ChIP-Seq screen. Top , gray vertical lines represent peak density plot from the
    Figure Legend Snippet: β-Catenin, TCF4, and RNAP bind the first intron of the YAP gene in vivo . A , diagram of the YAP gene locus and the β-catenin-enriched regions identified in a ChIP-Seq screen. Top , gray vertical lines represent peak density plot from the

    Techniques Used: In Vivo, Chromatin Immunoprecipitation

    Nuclear YAP expression is retained in densely grown CRC cells. A , Western blot analysis of YAP and β-catenin expression in six CRC cell lines. Tubulin served as a loading control. B , immunocytochemical analysis of YAP and β-catenin expression
    Figure Legend Snippet: Nuclear YAP expression is retained in densely grown CRC cells. A , Western blot analysis of YAP and β-catenin expression in six CRC cell lines. Tubulin served as a loading control. B , immunocytochemical analysis of YAP and β-catenin expression

    Techniques Used: Expressing, Western Blot

    The β-catenin/TCF4 binding region of YAP enhances SV40-driven luciferase gene expression. A , diagram of the pGL3-promoter plasmids used in luciferase assays with the SV40 minimal promoter and firefly luciferase gene shown as white rectangles and
    Figure Legend Snippet: The β-catenin/TCF4 binding region of YAP enhances SV40-driven luciferase gene expression. A , diagram of the pGL3-promoter plasmids used in luciferase assays with the SV40 minimal promoter and firefly luciferase gene shown as white rectangles and

    Techniques Used: Binding Assay, Luciferase, Expressing

    β-catenin and YAP are expressed in the nuclear compartment of cells comprising primary and metastatic colorectal tumors. A , immunohistochemical analysis of tissue arrays prepared from primary colorectal tumors, normal tissue, and colorectal tumors
    Figure Legend Snippet: β-catenin and YAP are expressed in the nuclear compartment of cells comprising primary and metastatic colorectal tumors. A , immunohistochemical analysis of tissue arrays prepared from primary colorectal tumors, normal tissue, and colorectal tumors

    Techniques Used: Immunohistochemistry

    β-Catenin controls YAP expression in HCT116 colon cancer cells. A , Western blot analysis of β-catenin protein levels in HCT116 cells expressing a control shRNA or an shRNA targeting β-catenin. HCT116 cells were infected with lentiviruses
    Figure Legend Snippet: β-Catenin controls YAP expression in HCT116 colon cancer cells. A , Western blot analysis of β-catenin protein levels in HCT116 cells expressing a control shRNA or an shRNA targeting β-catenin. HCT116 cells were infected with lentiviruses

    Techniques Used: Expressing, Western Blot, shRNA, Infection

    34) Product Images from "Fas and Fas Ligand Are Up-Regulated in Pulmonary Edema Fluid and Lung Tissue of Patients with Acute Lung Injury and the Acute Respiratory Distress Syndrome"

    Article Title: Fas and Fas Ligand Are Up-Regulated in Pulmonary Edema Fluid and Lung Tissue of Patients with Acute Lung Injury and the Acute Respiratory Distress Syndrome

    Journal: The American Journal of Pathology

    doi:

    Localization of markers of apoptosis in lung tissue sections from patients. The left column shows lung tissue sections from a patient who died with ALI or ARDS [identified as “(+) ARDS”]. The right column shows lung tissue sections from a patient who died without pulmonary disease [identified as “(−) ARDS”]. a to f: Tissue sections that are counterstained with hematoxylin. g to j: Tissue sections that were imaged using differential interference contrast optics, without counterstain, because the epithelial cell immunostain reaction product was subtle. The rows of pictures are matched for one marker of apoptosis: TUNEL ( a and b ), caspase-3 ( c and d ), Bax ( e and f ), Bcl II ( g and h ), and p53 ( i and j ). Cells lining, and in, the alveolar walls demonstrate more TUNEL-labeled nuclei ( arrow ), caspase-3-labeled cytoplasm ( arrow ), Bax-labeled cytoplasm ( arrow ), and p53-labeled cytoplasm ( arrow ) in the tissue sections from the patients who died with ALI or ARDS compared to the patient who died without pulmonary disease. On the other hand, Bcl II-labeled cells lining, and in, the alveolar walls are more prominent in the tissue sections from the patient who died without pulmonary disease compared to the patient who died with ALI or ARDS, as expected. All of the panels are the same magnification.
    Figure Legend Snippet: Localization of markers of apoptosis in lung tissue sections from patients. The left column shows lung tissue sections from a patient who died with ALI or ARDS [identified as “(+) ARDS”]. The right column shows lung tissue sections from a patient who died without pulmonary disease [identified as “(−) ARDS”]. a to f: Tissue sections that are counterstained with hematoxylin. g to j: Tissue sections that were imaged using differential interference contrast optics, without counterstain, because the epithelial cell immunostain reaction product was subtle. The rows of pictures are matched for one marker of apoptosis: TUNEL ( a and b ), caspase-3 ( c and d ), Bax ( e and f ), Bcl II ( g and h ), and p53 ( i and j ). Cells lining, and in, the alveolar walls demonstrate more TUNEL-labeled nuclei ( arrow ), caspase-3-labeled cytoplasm ( arrow ), Bax-labeled cytoplasm ( arrow ), and p53-labeled cytoplasm ( arrow ) in the tissue sections from the patients who died with ALI or ARDS compared to the patient who died without pulmonary disease. On the other hand, Bcl II-labeled cells lining, and in, the alveolar walls are more prominent in the tissue sections from the patient who died without pulmonary disease compared to the patient who died with ALI or ARDS, as expected. All of the panels are the same magnification.

    Techniques Used: Marker, TUNEL Assay, Labeling

    35) Product Images from "Urokinase-type plasminogen activator (uPA) is critical for progression of tuberous sclerosis complex 2 (TSC2)-deficient tumors"

    Article Title: Urokinase-type plasminogen activator (uPA) is critical for progression of tuberous sclerosis complex 2 (TSC2)-deficient tumors

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M117.799593

    Up-regulation of uPA in human LAM and angiomyolipoma tissue samples. A, normal human lung; B, human LAM nodules; C, normal human kidney; and D, renal angiomyolipoma. uPA was detected using a mouse anti-uPA monoclonal antibody (mAb) followed by Vectastain ABC combined with the Alexa Fluor 488-tyramide amplification system ( green ). SM α-actin was detected using Cy3-conjugated mouse SM α-actin mAb ( red ). Nuclei were counterstained with DAPI ( blue ) ( upper panels ). Total mouse IgG served as the negative controls for uPA staining, and Cy3-conjugated mouse anti-SM α-actin mAb was used to locate the same tumor nodules in serial sections ( bottom panels ). Additional serial sections were incubated with Cy3-conjugated total mouse IgG as the negative control for SM α-actin staining (data not shown). Images were taken using an LSM710 microscope. Scale bar, 50 μm. Data shown are representative of six LAM lesions from four patients and four angiomyolipomas from individual patients.
    Figure Legend Snippet: Up-regulation of uPA in human LAM and angiomyolipoma tissue samples. A, normal human lung; B, human LAM nodules; C, normal human kidney; and D, renal angiomyolipoma. uPA was detected using a mouse anti-uPA monoclonal antibody (mAb) followed by Vectastain ABC combined with the Alexa Fluor 488-tyramide amplification system ( green ). SM α-actin was detected using Cy3-conjugated mouse SM α-actin mAb ( red ). Nuclei were counterstained with DAPI ( blue ) ( upper panels ). Total mouse IgG served as the negative controls for uPA staining, and Cy3-conjugated mouse anti-SM α-actin mAb was used to locate the same tumor nodules in serial sections ( bottom panels ). Additional serial sections were incubated with Cy3-conjugated total mouse IgG as the negative control for SM α-actin staining (data not shown). Images were taken using an LSM710 microscope. Scale bar, 50 μm. Data shown are representative of six LAM lesions from four patients and four angiomyolipomas from individual patients.

    Techniques Used: Laser Capture Microdissection, Amplification, Staining, Incubation, Negative Control, Microscopy

    36) Product Images from "Abnormal Neuronal Migration Changes the Fate of Developing Neurons in the Postnatal Olfactory Bulb"

    Article Title: Abnormal Neuronal Migration Changes the Fate of Developing Neurons in the Postnatal Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.6716-10.2011

    Synaptic integration of Dcx ShRNA + cells in the core of the OB. A , Example of Dcx ShRNA + cells recorded at 12 and 30 dpe in the core of the OB. The presence of biocytin in the pipette solution confirmed the presence of GFP in the recorded cell and allowed visualization of cell position and morphology. Scale bar, 37 μm. B , Intrinsic membrane properties of control cells recorded at 12 dpe and Dcx ShRNA + cells at 12 and 30 dpe in the core of the OB. Membrane capacitance ( C m ), membrane resistance ( R m ), and maximum density of voltage-dependent K + channel ( I K + )- and Na + channel ( I Na + )-mediated currents were measured. Note that although the Dcx ShRNA + cells are blocked in the core of the OB, the changes of their intrinsic membrane properties confirm that they are undergoing maturation. C , Properties of spontaneous IPSCs at 0 mV received by Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Note that there is an increase of Dcx ShRNA + cells' spontaneous IPSC frequency, but not amplitude, over the maturation time. Calibration: 3 ms, 10 pA. D , Properties of IPSCs evoked by paired electrical stimulation in the GCL (voltage clamp = 0 mV; ISI = 150 ms) onto Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Note the increase of IPSC amplitude without modification of the paired-pulse ratio. Representative traces are averages of > 10 IPSCs. Calibration: 50 ms, 50 pA. E , Properties of EPSCs evoked by paired electrical stimulation in the GCL (voltage clamp = −70 mV; ISI = 50 ms) onto Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Representative traces are averages of > 10 IPSCs (calibration: 10 ms, 25 pA). Note the increase of IPSC amplitude and decrease of the paired-pulse ratio. Then, despite their ectopic location, Dcx ShRNA + cells undergo synaptic integration. Groups were compared 2 by 2 using a Mann–Whitney test (* p
    Figure Legend Snippet: Synaptic integration of Dcx ShRNA + cells in the core of the OB. A , Example of Dcx ShRNA + cells recorded at 12 and 30 dpe in the core of the OB. The presence of biocytin in the pipette solution confirmed the presence of GFP in the recorded cell and allowed visualization of cell position and morphology. Scale bar, 37 μm. B , Intrinsic membrane properties of control cells recorded at 12 dpe and Dcx ShRNA + cells at 12 and 30 dpe in the core of the OB. Membrane capacitance ( C m ), membrane resistance ( R m ), and maximum density of voltage-dependent K + channel ( I K + )- and Na + channel ( I Na + )-mediated currents were measured. Note that although the Dcx ShRNA + cells are blocked in the core of the OB, the changes of their intrinsic membrane properties confirm that they are undergoing maturation. C , Properties of spontaneous IPSCs at 0 mV received by Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Note that there is an increase of Dcx ShRNA + cells' spontaneous IPSC frequency, but not amplitude, over the maturation time. Calibration: 3 ms, 10 pA. D , Properties of IPSCs evoked by paired electrical stimulation in the GCL (voltage clamp = 0 mV; ISI = 150 ms) onto Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Note the increase of IPSC amplitude without modification of the paired-pulse ratio. Representative traces are averages of > 10 IPSCs. Calibration: 50 ms, 50 pA. E , Properties of EPSCs evoked by paired electrical stimulation in the GCL (voltage clamp = −70 mV; ISI = 50 ms) onto Dcx ShRNA + cells in the core of the OB at 12 and 30 dpe. Representative traces are averages of > 10 IPSCs (calibration: 10 ms, 25 pA). Note the increase of IPSC amplitude and decrease of the paired-pulse ratio. Then, despite their ectopic location, Dcx ShRNA + cells undergo synaptic integration. Groups were compared 2 by 2 using a Mann–Whitney test (* p

    Techniques Used: shRNA, Transferring, Mass Spectrometry, Modification, MANN-WHITNEY

    37) Product Images from "Role of OSGIN1 in mediating smoking-induced autophagy in the human airway epithelium"

    Article Title: Role of OSGIN1 in mediating smoking-induced autophagy in the human airway epithelium

    Journal: Autophagy

    doi: 10.1080/15548627.2017.1301327

    OSGIN1 induced autophagosome and autolysosome formation. Human primary airway basal cells were infected with lenti-control or lenti- OSGIN1 . Autophagosome and autolysosome formation was assessed by punctate staining of MAP1LC3B, using both immunofluorescence staining and immunohistochemistry staining. The effect of lenti- OSGIN1 at different doses, different times and in different subjects were quantified by the percentage of MAP1LC3B + cells. (A) Dose response. Left panels , immunofluorescence staining of MAP1LC3B in cells treated with the different dose of lenti-control and lenti- OSGIN1 . Blue, nuclei; red, MAP1LC3B; green, OSGIN1. Bar: 50 µm. Right panel , quantification of MAP1LC3B + cells. (B) Autophagic vacuole with MAP1LC3B staining. R, pseudo color red, MAP1LC3B; W, pseudo color white, OSGIN1; G, pseudo color green, GFP; B, pseudo color blue, nuclei; DIC, differential interference contrast. The inset is a combined image of DIC, MAP1LC3B and nuclei in an OSGIN1-expressing cell. Note the colocalization of MAP1LC3B with the vacuole structure in the cytoplasm. Bar: 10 µm. (C) Function of time. Left panels, immunohistochemistry staining of MAP1LC3B in cells at different time points after treatment (d 2 to d 5). Bottom insets are high magnification of MAP1LC3B staining from lenti- OSGIN1 , day 4 samples, demonstrating morphology of MAP1LC3B + cells. Blue, nuclei; red, MAP1LC3B. Bar: 50 µm. Bottom Right panel , quantification of MAP1LC3B + cells. (D) Different subjects. Quantification of MAP1LC3B + cells from experiments performed on 3 different subjects.
    Figure Legend Snippet: OSGIN1 induced autophagosome and autolysosome formation. Human primary airway basal cells were infected with lenti-control or lenti- OSGIN1 . Autophagosome and autolysosome formation was assessed by punctate staining of MAP1LC3B, using both immunofluorescence staining and immunohistochemistry staining. The effect of lenti- OSGIN1 at different doses, different times and in different subjects were quantified by the percentage of MAP1LC3B + cells. (A) Dose response. Left panels , immunofluorescence staining of MAP1LC3B in cells treated with the different dose of lenti-control and lenti- OSGIN1 . Blue, nuclei; red, MAP1LC3B; green, OSGIN1. Bar: 50 µm. Right panel , quantification of MAP1LC3B + cells. (B) Autophagic vacuole with MAP1LC3B staining. R, pseudo color red, MAP1LC3B; W, pseudo color white, OSGIN1; G, pseudo color green, GFP; B, pseudo color blue, nuclei; DIC, differential interference contrast. The inset is a combined image of DIC, MAP1LC3B and nuclei in an OSGIN1-expressing cell. Note the colocalization of MAP1LC3B with the vacuole structure in the cytoplasm. Bar: 10 µm. (C) Function of time. Left panels, immunohistochemistry staining of MAP1LC3B in cells at different time points after treatment (d 2 to d 5). Bottom insets are high magnification of MAP1LC3B staining from lenti- OSGIN1 , day 4 samples, demonstrating morphology of MAP1LC3B + cells. Blue, nuclei; red, MAP1LC3B. Bar: 50 µm. Bottom Right panel , quantification of MAP1LC3B + cells. (D) Different subjects. Quantification of MAP1LC3B + cells from experiments performed on 3 different subjects.

    Techniques Used: Infection, Staining, Immunofluorescence, Immunohistochemistry, Expressing

    38) Product Images from "Sialylation and fucosylation modulate inflammasome-activating eIF2 Signaling and microbial translocation during HIV infection"

    Article Title: Sialylation and fucosylation modulate inflammasome-activating eIF2 Signaling and microbial translocation during HIV infection

    Journal: Mucosal Immunology

    doi: 10.1038/s41385-020-0279-5

    The gut glycome is compartmentalized between the terminal ileum, right colon, and sigmoid colon of HIV+ ART-suppressed individuals. a A heat-map representing relative binding of various glycans to lectins in three sites of the gut from 20 HIV+ ART-suppressed individuals. Heat colors show standardized Z-scores across samples; red indicates higher binding levels and blue indicates lower binding levels. ANOVA test and Permanova test on Euclidean distance were used for statistical analysis. b Representative immunohistochemistry (IHC) staining using TJAII lectin (binds to α1,2 fucosylated glycans) on ileum, right colon, and sigmoid colon samples (5 patients for each site), ordered from left to right by levels of lectin binding as assessed by the lectin array. The numbers in the lower right corner are patient IDs. c Analysis of IHC staining as percentage of tissue area positive for α1,2 fucosylation (binding to TJAII) in the three sites. Statistical comparisons were performed using a Mann–Whitney test. Lines and error bars represent mean and standard error of the mean (SEM). d Correlation between IHC and lectin array data, shown as relative binding for TJAII using the lectin microarray and percentage of area positive for TJAII staining using IHC, evaluated using Spearman’s rank correlation coefficient test.
    Figure Legend Snippet: The gut glycome is compartmentalized between the terminal ileum, right colon, and sigmoid colon of HIV+ ART-suppressed individuals. a A heat-map representing relative binding of various glycans to lectins in three sites of the gut from 20 HIV+ ART-suppressed individuals. Heat colors show standardized Z-scores across samples; red indicates higher binding levels and blue indicates lower binding levels. ANOVA test and Permanova test on Euclidean distance were used for statistical analysis. b Representative immunohistochemistry (IHC) staining using TJAII lectin (binds to α1,2 fucosylated glycans) on ileum, right colon, and sigmoid colon samples (5 patients for each site), ordered from left to right by levels of lectin binding as assessed by the lectin array. The numbers in the lower right corner are patient IDs. c Analysis of IHC staining as percentage of tissue area positive for α1,2 fucosylation (binding to TJAII) in the three sites. Statistical comparisons were performed using a Mann–Whitney test. Lines and error bars represent mean and standard error of the mean (SEM). d Correlation between IHC and lectin array data, shown as relative binding for TJAII using the lectin microarray and percentage of area positive for TJAII staining using IHC, evaluated using Spearman’s rank correlation coefficient test.

    Techniques Used: Binding Assay, Immunohistochemistry, Staining, MANN-WHITNEY, Microarray

    39) Product Images from "α-catenin acts as a tumor suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signaling"

    Article Title: α-catenin acts as a tumor suppressor in E-cadherin-negative basal-like breast cancer by inhibiting NF-κB signaling

    Journal: Nature cell biology

    doi: 10.1038/ncb2909

    Loss of α-catenin promotes tumor growth in basal-like breast cancer cells by activating NF-κB signaling ( a ) Immunoblotting of α-catenin, RelA and HSP90 in MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. ( b ) Soft agar colony formation by MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. n = 4 wells per group. ( c ) Tumor growth by subcutaneously implanted MDA-MB-231 (3 × 10 6 cells injected) or BT549 (4 × 10 6 cells injected) cells infected with α-catenin shRNA alone or in combination with RelA shRNA. P values correspond to comparisons between α-catenin shRNA alone and α-catenin shRNA in combination with RelA shRNA. ( d, e ) Tumor weight ( d ) and tumor images ( e ) 5 weeks after mice were injected subcutaneously with MDA-MB-231 or BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. Scale bar: 1 cm. n = 5 (for MDA-MB-231 cells) or 8 (for BT549 cells) mice per group in ( c ) and ( d ). ( f, g ) TNFα ( f ) and human-specific IκBα ( g ) immunohistochemical staining of subcutaneous tumors formed by MDA-MB-231 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA, at 5 weeks after implantation. Scale bar: 50 μm. Data in ( b ) – ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: Loss of α-catenin promotes tumor growth in basal-like breast cancer cells by activating NF-κB signaling ( a ) Immunoblotting of α-catenin, RelA and HSP90 in MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. ( b ) Soft agar colony formation by MDA-MB-231 and BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. n = 4 wells per group. ( c ) Tumor growth by subcutaneously implanted MDA-MB-231 (3 × 10 6 cells injected) or BT549 (4 × 10 6 cells injected) cells infected with α-catenin shRNA alone or in combination with RelA shRNA. P values correspond to comparisons between α-catenin shRNA alone and α-catenin shRNA in combination with RelA shRNA. ( d, e ) Tumor weight ( d ) and tumor images ( e ) 5 weeks after mice were injected subcutaneously with MDA-MB-231 or BT549 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA. Scale bar: 1 cm. n = 5 (for MDA-MB-231 cells) or 8 (for BT549 cells) mice per group in ( c ) and ( d ). ( f, g ) TNFα ( f ) and human-specific IκBα ( g ) immunohistochemical staining of subcutaneous tumors formed by MDA-MB-231 cells transduced with α-catenin shRNA alone or in combination with RelA shRNA, at 5 weeks after implantation. Scale bar: 50 μm. Data in ( b ) – ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Multiple Displacement Amplification, Transduction, shRNA, Injection, Infection, Mouse Assay, Immunohistochemistry, Staining, Two Tailed Test

    α-catenin stabilizes IκBα protein by inhibiting IκBα ubiquitination and abrogating IκBα interaction with the proteasome ( a ) Upper panel: luciferase assays of NF-κB activity in MDA-MB-157 cells transfected with a pNFκB luciferase reporter alone or in combination with FLAG-α-catenin, with or without TNFα treatment. Lower panel: immunoblotting of FLAG, p-IκBα, IκBα, p-RelA, RelA and HSP90. n = 3 wells per group. ( b ) Immunoblotting of α-catenin, IκBα and HSP90 in α-catenin shRNA-transduced BT549 cells, with or without TNFα treatment. ( c ) Immunoblotting of IκBα and HSP90 in α-catenin-transduced MDA-MB-231 and MDA-MB-157 cells. ( d ) qPCR of CTNNA1 and NFKBIA in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( e ) α-catenin-transduced MDA-MB-157 cells were treated with 100 μg/ml of cycloheximide (CHX), harvested at different time points and immunoblotted with antibodies to α-catenin, IκBα and HSP90. ( f ) Quantification of IκBα protein levels in ( e ). ( g ) α-catenin was immunoprecipitated from α-catenin-transduced MDA-MB-157 cells and immunoblotted with antibodies to α-catenin and IκBα. ( h ) α-catenin was immunoprecipitated from MDA-MB-231 and BT549 cells and immunoblotted with antibodies to α-catenin, β-catenin, IκBα and RelA. ( i ) α-catenin was immunoprecipitated from untreated or TNFα-treated BT549 cells and immunoblotted with antibodies to α-catenin and IκBα. ( j ) α-catenin was immunoprecipitated from T47D or MCF10A cells and immunoblotted with antibodies to α-catenin, IκBα and RelA. ( k ) Upper panel: schematic representation of three vinculin domains of α-catenin. Lower panel: 293T cells were cotransfected with MYC-IκBα and SFB-tagged full-length α-catenin or fragment VH1, VH2 or VH3. α-catenin and the three fragments were purified with S-protein beads and immunoblotted with antibodies to FLAG and MYC. ( l ) HA-tagged wild-type ubiquitin (Ub) or the K48R or K63R mutant was cotransfected with MYC-α-catenin and SFB-IκBα into 293T cells. Cells were treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to HA and FLAG. ( m ) 293T cells were cotransfected with MYC-α-catenin and SFB-IκBα and then treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to the 20S proteasome subunit α4, MYC and FLAG. Data in ( a ) and ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: α-catenin stabilizes IκBα protein by inhibiting IκBα ubiquitination and abrogating IκBα interaction with the proteasome ( a ) Upper panel: luciferase assays of NF-κB activity in MDA-MB-157 cells transfected with a pNFκB luciferase reporter alone or in combination with FLAG-α-catenin, with or without TNFα treatment. Lower panel: immunoblotting of FLAG, p-IκBα, IκBα, p-RelA, RelA and HSP90. n = 3 wells per group. ( b ) Immunoblotting of α-catenin, IκBα and HSP90 in α-catenin shRNA-transduced BT549 cells, with or without TNFα treatment. ( c ) Immunoblotting of IκBα and HSP90 in α-catenin-transduced MDA-MB-231 and MDA-MB-157 cells. ( d ) qPCR of CTNNA1 and NFKBIA in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( e ) α-catenin-transduced MDA-MB-157 cells were treated with 100 μg/ml of cycloheximide (CHX), harvested at different time points and immunoblotted with antibodies to α-catenin, IκBα and HSP90. ( f ) Quantification of IκBα protein levels in ( e ). ( g ) α-catenin was immunoprecipitated from α-catenin-transduced MDA-MB-157 cells and immunoblotted with antibodies to α-catenin and IκBα. ( h ) α-catenin was immunoprecipitated from MDA-MB-231 and BT549 cells and immunoblotted with antibodies to α-catenin, β-catenin, IκBα and RelA. ( i ) α-catenin was immunoprecipitated from untreated or TNFα-treated BT549 cells and immunoblotted with antibodies to α-catenin and IκBα. ( j ) α-catenin was immunoprecipitated from T47D or MCF10A cells and immunoblotted with antibodies to α-catenin, IκBα and RelA. ( k ) Upper panel: schematic representation of three vinculin domains of α-catenin. Lower panel: 293T cells were cotransfected with MYC-IκBα and SFB-tagged full-length α-catenin or fragment VH1, VH2 or VH3. α-catenin and the three fragments were purified with S-protein beads and immunoblotted with antibodies to FLAG and MYC. ( l ) HA-tagged wild-type ubiquitin (Ub) or the K48R or K63R mutant was cotransfected with MYC-α-catenin and SFB-IκBα into 293T cells. Cells were treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to HA and FLAG. ( m ) 293T cells were cotransfected with MYC-α-catenin and SFB-IκBα and then treated with 10 μM MG132 and 20 ng/ml of TNFα for 30 minutes. IκBα was purified with S-protein beads and immunoblotted with antibodies to the 20S proteasome subunit α4, MYC and FLAG. Data in ( a ) and ( d ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Luciferase, Activity Assay, Multiple Displacement Amplification, Transfection, shRNA, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Mutagenesis, Two Tailed Test

    α-catenin inhibits RelA-p50 nuclear localization and downregulates RelB ( a ) Immunoblotting of α-catenin and RelA in cytoplasmic and nuclear fractions of BT549 cells transduced with two independent α-catenin shRNAs, with or without TNFα treatment. ( b ) Immunoblotting of α-catenin, p105, p50, RelA, RelB and IκBα in cytoplasmic and nuclear fractions of α-catenin-transduced MDA-MB-157 cells. α-tubulin and Lamin A were used as cytoplasmic and nuclear markers, respectively, in ( a ) and ( b ). ( c ) Schematic representation of the RELB promoter containing two RelA binding sites (red rectangles). The two boxed arrows indicate the primers used for ChIP-qPCR. Hs: Homo sapiens , Mm: Mus musculus . ( d ) ChIP-qPCR analysis of RelA binding to the RELB promoter in α-catenin-transduced MDA-MB-157 cells. qPCR was performed with primers specific to the RelA binding motifs. Data were normalized to the input. n = 3 samples per group. ( e ) qPCR of RELB in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( f ) Immunoblotting of RelB and HSP90 in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. SE: short exposure; LE: long exposure. Data in ( d ) and ( e ) are the mean of biological replicates from a representative experiment,, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: α-catenin inhibits RelA-p50 nuclear localization and downregulates RelB ( a ) Immunoblotting of α-catenin and RelA in cytoplasmic and nuclear fractions of BT549 cells transduced with two independent α-catenin shRNAs, with or without TNFα treatment. ( b ) Immunoblotting of α-catenin, p105, p50, RelA, RelB and IκBα in cytoplasmic and nuclear fractions of α-catenin-transduced MDA-MB-157 cells. α-tubulin and Lamin A were used as cytoplasmic and nuclear markers, respectively, in ( a ) and ( b ). ( c ) Schematic representation of the RELB promoter containing two RelA binding sites (red rectangles). The two boxed arrows indicate the primers used for ChIP-qPCR. Hs: Homo sapiens , Mm: Mus musculus . ( d ) ChIP-qPCR analysis of RelA binding to the RELB promoter in α-catenin-transduced MDA-MB-157 cells. qPCR was performed with primers specific to the RelA binding motifs. Data were normalized to the input. n = 3 samples per group. ( e ) qPCR of RELB in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. n = 3 samples per group. ( f ) Immunoblotting of RelB and HSP90 in α-catenin-transduced MDA-MB-157 and MDA-MB-436 cells. SE: short exposure; LE: long exposure. Data in ( d ) and ( e ) are the mean of biological replicates from a representative experiment,, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Transduction, Multiple Displacement Amplification, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Two Tailed Test

    α-catenin inhibits tumorigenesis and is downregulated in human basal-like breast cancer ( a ) Immunoblotting of α-catenin, IκBα, RelB and HSP90 in MDA-MB-157 cells transduced with α-catenin alone or in combination with IκBα shRNA. ( b, c ) Images ( b ) and quantification ( c ) of growth curves of cells described in ( a ). n = 3 wells per group. ( d, e ) Images ( d ) and quantification ( e ) of soft agar colony formation by cells described in ( a ). Scale bar: 100 μm. n = 5 wells per group. ( f ) Tumor growth by 3 × 10 6 subcutaneously injected cells described in ( a ). P values correspond to comparisons between α-catenin alone and α-catenin in combination with IκBα shRNA (shIκBα-3) in ( c ) and ( f ). ( g, h ) Tumor weight ( g ) and tumor images ( h ) of mice described in ( f ). Scale bar: 1cm. n = 5 mice per group in ( f ) and ( g ). ( i ) Box plots comparing CTNNA1 expression in normal breast tissues and in total ( n = 59) and basal-like ( n = 10) breast tumors. Statistical significance was determined by the Wilcoxon test. The boxes show the median and the interquartile range. The whiskers show the minimum and maximum. ( j ) Kaplan-Meier curves of relapse-free survival times of total breast cancer patients ( n = 2878) and patients with basal-like breast cancer ( n = 478), stratified by CTNNA1 expression levels. Data were obtained from http://kmplot.com/analysis/ 34 . Statistical significance was determined by the log-rank test. ( k ) Scatterplots showing the inverse correlation of CTNNA1 with TNF (left panel) or RELB (right panel) expression in human breast tumors ( n = 805). ( l ) Scatterplots showing the inverse correlation between methylation of the CTNNA1 gene and CTNNA1 expression in total (left panel, n = 511) and basal-like (right panel, n = 39) breast tumors. Statistical significance in ( k ) and ( l ) was determined by Spearman rank correlation test. Rs = Spearman rank correlation coefficient. Data in ( c ) and ( e ) – ( g ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .
    Figure Legend Snippet: α-catenin inhibits tumorigenesis and is downregulated in human basal-like breast cancer ( a ) Immunoblotting of α-catenin, IκBα, RelB and HSP90 in MDA-MB-157 cells transduced with α-catenin alone or in combination with IκBα shRNA. ( b, c ) Images ( b ) and quantification ( c ) of growth curves of cells described in ( a ). n = 3 wells per group. ( d, e ) Images ( d ) and quantification ( e ) of soft agar colony formation by cells described in ( a ). Scale bar: 100 μm. n = 5 wells per group. ( f ) Tumor growth by 3 × 10 6 subcutaneously injected cells described in ( a ). P values correspond to comparisons between α-catenin alone and α-catenin in combination with IκBα shRNA (shIκBα-3) in ( c ) and ( f ). ( g, h ) Tumor weight ( g ) and tumor images ( h ) of mice described in ( f ). Scale bar: 1cm. n = 5 mice per group in ( f ) and ( g ). ( i ) Box plots comparing CTNNA1 expression in normal breast tissues and in total ( n = 59) and basal-like ( n = 10) breast tumors. Statistical significance was determined by the Wilcoxon test. The boxes show the median and the interquartile range. The whiskers show the minimum and maximum. ( j ) Kaplan-Meier curves of relapse-free survival times of total breast cancer patients ( n = 2878) and patients with basal-like breast cancer ( n = 478), stratified by CTNNA1 expression levels. Data were obtained from http://kmplot.com/analysis/ 34 . Statistical significance was determined by the log-rank test. ( k ) Scatterplots showing the inverse correlation of CTNNA1 with TNF (left panel) or RELB (right panel) expression in human breast tumors ( n = 805). ( l ) Scatterplots showing the inverse correlation between methylation of the CTNNA1 gene and CTNNA1 expression in total (left panel, n = 511) and basal-like (right panel, n = 39) breast tumors. Statistical significance in ( k ) and ( l ) was determined by Spearman rank correlation test. Rs = Spearman rank correlation coefficient. Data in ( c ) and ( e ) – ( g ) are the mean of biological replicates from a representative experiment, and error bars indicate s.e.m. Statistical significance was determined by a two-tailed, unpaired Student’s t -test. The experiments were repeated three times. The source data can be found in Supplementary Table 4 . Uncropped images of blots are shown in Supplementary Fig. 7 .

    Techniques Used: Multiple Displacement Amplification, Transduction, shRNA, Injection, Mouse Assay, Expressing, Methylation, Two Tailed Test

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

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

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086892

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

    Techniques Used: Expressing, Microarray

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

    Techniques Used: Expressing, Microarray

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    Next-Generation Sequencing:

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

    Article Title: Combinational therapy using hypothermia and the immunophilin ligand FK506 to target altered pial arteriolar reactivity, axonal damage, and blood-brain barrier dysfunction after traumatic brain injury in rat
    Article Snippet: .. Next, the sections were incubated for 1 hour with biotinylated goat anti-rabbit immunoglobulin G (IgG) (Vector Laboratories Inc., Burlingame, CA, USA) diluted 1:1,000 in 1% normal goat serum in PBS. .. The reaction product was visualized by incubation for 1 hour in avidin-biotinylated enzyme complex (Vectastain ABC kit, Vector Laboratories Inc.), followed by 0.05% diaminobenzidene, 0.01% H2 O2 , and 0.3% imidazole in 0.1% mL sodium phosphate buffer for 15 minutes.

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    Vector Laboratories vectastain elite abc hrp kit peroxidase standard
    Specificity of WFA labeling of PNN in the BLA of weanling rats. Panel a: <t>HRP-DAB</t> was used to detect WFA labeling. Specificity of WFA labeling was confirmed by reduction of WFA labeling within the brains injected with the enzymes, <t>chondroitinase-ABC</t> and hyaluronidase, known to dissolve proteoglycans of the PNN and surrounding neuropil. The region infused with chondroitinase-ABC and hyaluronidase exhibited complete absence of the HRP-DAB reaction product, while PNN labeling remained intensely labeled in the surrounding regions (e.g., dorsal hippocampus and reticular thalamus) (scale bar= 200 µm). Panel b : The pink contour indicates the boundary of the BLA (scale bar = 200 µm). Panel c : The lower right panel shows detail of the enzyme-infused region (scale bar = 50 µm). All panels were taken at a magnification of 10X.
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    Specificity of WFA labeling of PNN in the BLA of weanling rats. Panel a: HRP-DAB was used to detect WFA labeling. Specificity of WFA labeling was confirmed by reduction of WFA labeling within the brains injected with the enzymes, chondroitinase-ABC and hyaluronidase, known to dissolve proteoglycans of the PNN and surrounding neuropil. The region infused with chondroitinase-ABC and hyaluronidase exhibited complete absence of the HRP-DAB reaction product, while PNN labeling remained intensely labeled in the surrounding regions (e.g., dorsal hippocampus and reticular thalamus) (scale bar= 200 µm). Panel b : The pink contour indicates the boundary of the BLA (scale bar = 200 µm). Panel c : The lower right panel shows detail of the enzyme-infused region (scale bar = 50 µm). All panels were taken at a magnification of 10X.

    Journal: The Journal of comparative neurology

    Article Title: Early life trauma increases threat response of peri-weaning rats, reduction of axo-somatic synapses formed by parvalbumin cells and perineuronal net in the basolateral nucleus of amygdala

    doi: 10.1002/cne.24522

    Figure Lengend Snippet: Specificity of WFA labeling of PNN in the BLA of weanling rats. Panel a: HRP-DAB was used to detect WFA labeling. Specificity of WFA labeling was confirmed by reduction of WFA labeling within the brains injected with the enzymes, chondroitinase-ABC and hyaluronidase, known to dissolve proteoglycans of the PNN and surrounding neuropil. The region infused with chondroitinase-ABC and hyaluronidase exhibited complete absence of the HRP-DAB reaction product, while PNN labeling remained intensely labeled in the surrounding regions (e.g., dorsal hippocampus and reticular thalamus) (scale bar= 200 µm). Panel b : The pink contour indicates the boundary of the BLA (scale bar = 200 µm). Panel c : The lower right panel shows detail of the enzyme-infused region (scale bar = 50 µm). All panels were taken at a magnification of 10X.

    Article Snippet: Tissue was then washed in 0.01M PBS, incubated in a solution of the Vectastain Elite ABC HRP kit (Vector Laboratories, Burlingame, CA, cat# PK-6100), washed with 0.01M PBS, and incubated for 9 min in a filtered solution of 3’3-diaminobenzidine tetrahydrochloride (DAB; 10mg tablet Sigma Aldrich in 44ml of PBS buffer), catalyzed by 0.003% hydrogen peroxide.

    Techniques: Labeling, Injection

    Histopathological findings in the liver of a Rousettus aegyptiacus fruit bat immunized with the MP-12 vaccine strain at day seven post immunization. ( A ) Histopathology shows few, randomly distributed foci of hepatocellular necrosis and loss with macrophage and lymphocyte infiltration (arrow). Furthermore, the hepatocytes display moderate, coalescing to diffuse, floccular cytoplasmic vacuolization, interpreted as a species-specific, relatively high level of glycogen storage. Hematoxylin-eosin. Bar = 100 μm; ( B ) Immunohistochemistry for Rift Valley fever phlebovirus (RVFV) Gc antigen reveals minor amounts of intra- and extracellular, strongly immunoreactive granula within the lesions (arrow), interpreted as debris remaining after virus-induced hepatocellular death. Immunohistochemistry, monoclonal mouse anti-RVFV Gc-protein antibody, avidin-biotin-peroxidase-complex method, 3-amino-9-ethyl-carbazol chromogen (red), hematoxylin counterstain (blue). Bar = 20 μm.

    Journal: Viruses

    Article Title: Productive Propagation of Rift Valley Fever Phlebovirus Vaccine Strain MP-12 in Rousettus aegyptiacus Fruit Bats

    doi: 10.3390/v10120681

    Figure Lengend Snippet: Histopathological findings in the liver of a Rousettus aegyptiacus fruit bat immunized with the MP-12 vaccine strain at day seven post immunization. ( A ) Histopathology shows few, randomly distributed foci of hepatocellular necrosis and loss with macrophage and lymphocyte infiltration (arrow). Furthermore, the hepatocytes display moderate, coalescing to diffuse, floccular cytoplasmic vacuolization, interpreted as a species-specific, relatively high level of glycogen storage. Hematoxylin-eosin. Bar = 100 μm; ( B ) Immunohistochemistry for Rift Valley fever phlebovirus (RVFV) Gc antigen reveals minor amounts of intra- and extracellular, strongly immunoreactive granula within the lesions (arrow), interpreted as debris remaining after virus-induced hepatocellular death. Immunohistochemistry, monoclonal mouse anti-RVFV Gc-protein antibody, avidin-biotin-peroxidase-complex method, 3-amino-9-ethyl-carbazol chromogen (red), hematoxylin counterstain (blue). Bar = 20 μm.

    Article Snippet: Immunohistology was performed using a mouse monoclonal antibody against the RVFV Gc-protein (clone: GC9A9) [ ], the avidin–biotin–peroxidase complex method (ABC, Elite PK6100; Vector Laboratories, Burlingame, CA, USA) with 3-amino-9-ethylcarbazole (AEC, Dako, Glostrup, Denmark) as chromogen and hematoxylin counterstain.

    Techniques: Histopathology, Immunohistochemistry, Avidin-Biotin Assay

    Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.

    Journal: Emerging Microbes & Infections

    Article Title: Modulation of lethal HPAIV H5N8 clade 2.3.4.4B infection in AIV pre-exposed mallards

    doi: 10.1080/22221751.2020.1713706

    Figure Lengend Snippet: Light microscopic finding in the livers of experimentally H5N8B-infected ducks. (A, B) Seropositive mallard, H5N8B-infected, clinically normal, 34 dpi, liver. (A) No obvious findings. (B) Lack of immunohistochemically-detectable hepatocellular influenza A virus matrixprotein antigen. (C, D) Pekin duck, contact animal, died 4 days post contact, liver. (C) Marked hypereosinophilia, hepatocellular vacuolation, membraneous rupture and nuclear pyknosis, karyorrhexis and lysis interpreted as severe, acute, coalescing to diffuse necrotizing hepatitis. (D) Immunohistochemistry reveals coalescing intrahepatocytic, intracytoplasmic and intranuclear influenza A virus matrix protein. (A, C) Hematoxylin-eosin, (B, D) Immunohistochemistry using the avidin-biotin-peroxidase-complex method with a monoclonal antibody against influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (redbrown) and hematoxylin counterstain (blue). (A, C) bars = 20 μm. (B, D) bars = 50 μm.

    Article Snippet: Immunohistochemistry was employed to detect influenza A virus matrix protein using the avidin–biotin-peroxidase-complex method (Vectastain PK 6100; Vector Laboratories, Burlingame, CA, USA) with citric buffer (10 mM, pH 6.0) pretreatment, a monoclonal antibody (mAb) directed against an epitope of the influenza A virus matrix protein (ATCC clone HB-64), 3-amino-9-ethylcarbazol chromogen (Agilent Technologies, Santa Clara, CA, USA), and hematoxylin counterstain [ ].

    Techniques: Infection, Lysis, Immunohistochemistry, Avidin-Biotin Assay

    The ABC peroxidase technique. This approach is similar to that shown in , but substitutes a biotinylated peroxidase. When incubated with a substrate such as DAB (lower left) or NiDAB (lower right), the colored insoluble product is deposited

    Journal: Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.]

    Article Title: The Importance of Titrating Antibodies for Immunocytochemical Methods

    doi: 10.1002/cpns.1

    Figure Lengend Snippet: The ABC peroxidase technique. This approach is similar to that shown in , but substitutes a biotinylated peroxidase. When incubated with a substrate such as DAB (lower left) or NiDAB (lower right), the colored insoluble product is deposited

    Article Snippet: Brain sections as obtained in Basic Protocol 1, steps 1 to 15 Primary antibody Biotinylated secondary antibody Vectastain Elite ABC Kit (Standard; Vector Laboratories, cat. no. PK-6100) including solution A and Solution B Biotinylated tyramine produced as outlined in .

    Techniques: Incubation

    TSA-amplified fluorescence using biotinylated tyramine and streptavidin fluorophore. Note the greatly increased number of fluorescent molecules compared with that seen for either fluorophore-tagged secondaries or ABC streptavidin methods. P = peroxidase.

    Journal: Current protocols in neuroscience / editorial board, Jacqueline N. Crawley ... [et al.]

    Article Title: The Importance of Titrating Antibodies for Immunocytochemical Methods

    doi: 10.1002/cpns.1

    Figure Lengend Snippet: TSA-amplified fluorescence using biotinylated tyramine and streptavidin fluorophore. Note the greatly increased number of fluorescent molecules compared with that seen for either fluorophore-tagged secondaries or ABC streptavidin methods. P = peroxidase.

    Article Snippet: Brain sections as obtained in Basic Protocol 1, steps 1 to 15 Primary antibody Biotinylated secondary antibody Vectastain Elite ABC Kit (Standard; Vector Laboratories, cat. no. PK-6100) including solution A and Solution B Biotinylated tyramine produced as outlined in .

    Techniques: Amplification, Fluorescence