immunohistochemical analysis immunostaining  (Vector Laboratories)


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    Streptavidin Horseradish Peroxidase Concentrate for IHC
    Description:
    Horseradish Peroxidase Streptavidin Concentrate for IHC are powerful tools to detect or purify biotinylated proteins nucleic acids and other macromolecules Horseradish Peroxidase Streptavidin HRP Streptavidin is produced by our own coupling procedure which preserves the high specific activity of the peroxidase Using biotinylated primary or secondary intermediates and peroxidase labeled streptavidin antigens can be localized in histological sections cytospin preparations or smears Horseradish Peroxidase Streptavidin is supplied as a concentrate 1mg ml or as a ready to use R T U stabilized solution 100 ml 1 g ml in a bottle fitted with a drop dispenser top For horseradish peroxidase streptavidin Sav HRP that is optimized for use in solid phase assays such as ELISAs and blotting applications see the following product Streptavidin Horseradish Peroxidase Sav HRP Concentrate for ELISAs and Blots
    Catalog Number:
    sa-5004
    Price:
    None
    Size:
    1 mg
    Category:
    Protein chemifluorescent detection reagents or kits or substrates
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    Structured Review

    Vector Laboratories immunohistochemical analysis immunostaining
    Streptavidin Horseradish Peroxidase Concentrate for IHC
    Horseradish Peroxidase Streptavidin Concentrate for IHC are powerful tools to detect or purify biotinylated proteins nucleic acids and other macromolecules Horseradish Peroxidase Streptavidin HRP Streptavidin is produced by our own coupling procedure which preserves the high specific activity of the peroxidase Using biotinylated primary or secondary intermediates and peroxidase labeled streptavidin antigens can be localized in histological sections cytospin preparations or smears Horseradish Peroxidase Streptavidin is supplied as a concentrate 1mg ml or as a ready to use R T U stabilized solution 100 ml 1 g ml in a bottle fitted with a drop dispenser top For horseradish peroxidase streptavidin Sav HRP that is optimized for use in solid phase assays such as ELISAs and blotting applications see the following product Streptavidin Horseradish Peroxidase Sav HRP Concentrate for ELISAs and Blots
    https://www.bioz.com/result/immunohistochemical analysis immunostaining/product/Vector Laboratories
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    Price from $9.99 to $1999.99
    immunohistochemical analysis immunostaining - by Bioz Stars, 2021-02
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    Images

    1) Product Images from "Follistatin, an Activin Antagonist, Ameliorates Renal Interstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction"

    Article Title: Follistatin, an Activin Antagonist, Ameliorates Renal Interstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction

    Journal: BioMed Research International

    doi: 10.1155/2014/376191

    Effects of follistatin on expression of α -SMA in kidneys after UUO. (a) Expression of α -SMA, a marker for myofibroblasts, in the UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 10 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 10 days. (F) Follistatin-treated UUO kidneys, 10 days. Magnification: ×200. α -SMA (green), DAPI (blue). (b) Quantitative analysis of α -SMA-positive area. α -SMA-positive area in the kidneys at 10 days after UUO was assessed as described in Section 2 . Values are mean ± SE ( n = 5). * P
    Figure Legend Snippet: Effects of follistatin on expression of α -SMA in kidneys after UUO. (a) Expression of α -SMA, a marker for myofibroblasts, in the UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 10 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 10 days. (F) Follistatin-treated UUO kidneys, 10 days. Magnification: ×200. α -SMA (green), DAPI (blue). (b) Quantitative analysis of α -SMA-positive area. α -SMA-positive area in the kidneys at 10 days after UUO was assessed as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Techniques Used: Expressing, Marker, Immunostaining

    Effects of follistatin on the production of extracellular matrix in kidneys after UUO. (a) Production of type I collagen (A–D), type III collagen (E–H), and fibronectin (I–L) in the UUO kidneys was examined by immunostaining. (A, E, I) normal kidney. (B, F, J) contralateral kidney. (C, G, K) saline-treated UUO kidney, 7 days. (D, H, L) follistatin-treated UUO kidney, 7 days. Type I collagen, type III collagen, and fibronectin (green). Magnification: ×100. (b) Quantitative analysis of extracellular matrix production. Type I collagen, type III collagen, and fibronectin-positive area in kidneys at 7 days after UUO was measured as described in Section 2 . Values are mean ± SE ( n = 5). * P
    Figure Legend Snippet: Effects of follistatin on the production of extracellular matrix in kidneys after UUO. (a) Production of type I collagen (A–D), type III collagen (E–H), and fibronectin (I–L) in the UUO kidneys was examined by immunostaining. (A, E, I) normal kidney. (B, F, J) contralateral kidney. (C, G, K) saline-treated UUO kidney, 7 days. (D, H, L) follistatin-treated UUO kidney, 7 days. Type I collagen, type III collagen, and fibronectin (green). Magnification: ×100. (b) Quantitative analysis of extracellular matrix production. Type I collagen, type III collagen, and fibronectin-positive area in kidneys at 7 days after UUO was measured as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Techniques Used: Immunostaining

    Effects of follistatin on macrophage infiltration in kidneys after UUO. (a) Expression of CD68, a marker for macrophages, in UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 3 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 7 days. (F) Follistatin-treated UUO kidneys, 7 days. CD68-positive cells (brown). Magnification: ×400. (b) Quantitative analysis of CD68-positive cell number. CD68-positive cells were counted in 10 randomly selected fields per rat at ×400 magnification. Values are mean ± SE ( n = 5). Saline (white bars), follistatin (black bars). * P
    Figure Legend Snippet: Effects of follistatin on macrophage infiltration in kidneys after UUO. (a) Expression of CD68, a marker for macrophages, in UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 3 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 7 days. (F) Follistatin-treated UUO kidneys, 7 days. CD68-positive cells (brown). Magnification: ×400. (b) Quantitative analysis of CD68-positive cell number. CD68-positive cells were counted in 10 randomly selected fields per rat at ×400 magnification. Values are mean ± SE ( n = 5). Saline (white bars), follistatin (black bars). * P

    Techniques Used: Expressing, Marker, Immunostaining

    2) Product Images from "Therapeutic Vaccination against Helicobacter pylori in the Beagle Dog Experimental Model: Safety, Immunogenicity, and Efficacy "

    Article Title: Therapeutic Vaccination against Helicobacter pylori in the Beagle Dog Experimental Model: Safety, Immunogenicity, and Efficacy

    Journal:

    doi: 10.1128/IAI.72.6.3252-3259.2004

    Study 2 (monthly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (week 23 postvaccination). The higher magnification
    Figure Legend Snippet: Study 2 (monthly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (week 23 postvaccination). The higher magnification

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Sampling

    Study 1 (weekly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (29th week postvaccination). The higher magnification
    Figure Legend Snippet: Study 1 (weekly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (29th week postvaccination). The higher magnification

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Sampling

    3) Product Images from "Butyrate and bioactive proteolytic form of Wnt-5a regulate colonic epithelial proliferation and spatial development"

    Article Title: Butyrate and bioactive proteolytic form of Wnt-5a regulate colonic epithelial proliferation and spatial development

    Journal: Scientific Reports

    doi: 10.1038/srep32094

    Butyrate inhibition of colonic epithelial cell growth is dependent on induction of Hsp25. ( a ) Butyrate inhibition of cell growth (clear bar, fourth from left) is blocked by silencing Hsp25 (grey bar, far right), but not by scrambled (non-sense) siRNA (siScr). Cell proliferation was measured by the WST-1 assay. (b) Lentiviral-induced expression of Hsp25 (Hsp25 lenti) in mouse colon decreases expression of the proliferation marker Ki-67 (brown staining). As a control, empty cassette GFP lentivector (GFP-lenti) was administered the same way. After one hour exposure of the colonic mucosa to luminally-administered lentivirus, the colonic mucosal segment was marked, returned to the abdominal cavity, and then harvested 3 days later, as described in methods. (c) Upper panel : Alpha smooth muscle actin staining was used to identify peri-crypt myofibroblasts (brown immunostaining). Lower panel: Distribution of pericrypt myofibroblast, identified by alpha smooth muscle actin immunostaining, among the upper, middle, and lower tritiles (thirds) of colonic crypts. (d) Intestinal myofibroblast-derived conditioned medium (CM) blocks butyrate-stimulated (1 and 5 mM) Hsp25 expression in intestinal epithelial YAMC monolayers. No effects were seen on Hsc70 protein expression which is used as a loading control (d, upper panel). *p
    Figure Legend Snippet: Butyrate inhibition of colonic epithelial cell growth is dependent on induction of Hsp25. ( a ) Butyrate inhibition of cell growth (clear bar, fourth from left) is blocked by silencing Hsp25 (grey bar, far right), but not by scrambled (non-sense) siRNA (siScr). Cell proliferation was measured by the WST-1 assay. (b) Lentiviral-induced expression of Hsp25 (Hsp25 lenti) in mouse colon decreases expression of the proliferation marker Ki-67 (brown staining). As a control, empty cassette GFP lentivector (GFP-lenti) was administered the same way. After one hour exposure of the colonic mucosa to luminally-administered lentivirus, the colonic mucosal segment was marked, returned to the abdominal cavity, and then harvested 3 days later, as described in methods. (c) Upper panel : Alpha smooth muscle actin staining was used to identify peri-crypt myofibroblasts (brown immunostaining). Lower panel: Distribution of pericrypt myofibroblast, identified by alpha smooth muscle actin immunostaining, among the upper, middle, and lower tritiles (thirds) of colonic crypts. (d) Intestinal myofibroblast-derived conditioned medium (CM) blocks butyrate-stimulated (1 and 5 mM) Hsp25 expression in intestinal epithelial YAMC monolayers. No effects were seen on Hsc70 protein expression which is used as a loading control (d, upper panel). *p

    Techniques Used: Inhibition, WST-1 Assay, Expressing, Marker, Staining, Immunostaining, Derivative Assay

    4) Product Images from "Identification of Urinary Activin A as a Novel Biomarker Reflecting the Severity of Acute Kidney Injury"

    Article Title: Identification of Urinary Activin A as a Novel Biomarker Reflecting the Severity of Acute Kidney Injury

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-23564-3

    Localization of A subunit for Activin, NGAL, and KIM-1 in the Kidneys after Renal Ischemia. ( A ) Localization of βA subunit for activin and NGAL in the kidneys after renal ischemia for 25 min was examined by immunostaining. βA subunit (green), NGAL (red). Magnification: ×400. ( B ) Localization of βA subunit for activin and KIM-1 in the kidneys after renal ischemia was examined by immunostaining. βA subunit (green), KIM-1 (red). Magnification: ×1000. ( C ) Localization of βA subunit for activin and PCNA in the ischemic kidneys at 48 hr after reperfusion. βA subunit (red), PCNA (green), and DAPI (blue). Magnification: ×400. ( D ) Immunostaining of βA subunit for activin (a, b) and TUNEL staining (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’).Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000. ( E ) Immunostaining of βA subunit for activin (a,b) and caspase 3 (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’). Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000.
    Figure Legend Snippet: Localization of A subunit for Activin, NGAL, and KIM-1 in the Kidneys after Renal Ischemia. ( A ) Localization of βA subunit for activin and NGAL in the kidneys after renal ischemia for 25 min was examined by immunostaining. βA subunit (green), NGAL (red). Magnification: ×400. ( B ) Localization of βA subunit for activin and KIM-1 in the kidneys after renal ischemia was examined by immunostaining. βA subunit (green), KIM-1 (red). Magnification: ×1000. ( C ) Localization of βA subunit for activin and PCNA in the ischemic kidneys at 48 hr after reperfusion. βA subunit (red), PCNA (green), and DAPI (blue). Magnification: ×400. ( D ) Immunostaining of βA subunit for activin (a, b) and TUNEL staining (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’).Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000. ( E ) Immunostaining of βA subunit for activin (a,b) and caspase 3 (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’). Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000.

    Techniques Used: Immunostaining, TUNEL Assay, Staining

    5) Product Images from "Cripto-1 Ablation Disrupts Alveolar Development in the Mouse Mammary Gland through a Progesterone Receptor–Mediated Pathway"

    Article Title: Cripto-1 Ablation Disrupts Alveolar Development in the Mouse Mammary Gland through a Progesterone Receptor–Mediated Pathway

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.07.023

    Effects of early and late deletion of mouse Cripto-1 on progesterone receptor (PR) expression. Representative PR-positive cells. A: Immunohistochemical analysis of mammary tissue collected from FVB/N, whey acidic protein- Cre + / Cripto flox (WAP-CRKO), and
    Figure Legend Snippet: Effects of early and late deletion of mouse Cripto-1 on progesterone receptor (PR) expression. Representative PR-positive cells. A: Immunohistochemical analysis of mammary tissue collected from FVB/N, whey acidic protein- Cre + / Cripto flox (WAP-CRKO), and

    Techniques Used: Expressing, Immunohistochemistry

    Effects of early and late deletion of mouse Cripto-1 on proliferation. Proliferation-positive cells were scored and labeling index expressed as a percentage of positive nuclei from at least 3000 counted cells in mammary glands. A: Representative immunohistochemical
    Figure Legend Snippet: Effects of early and late deletion of mouse Cripto-1 on proliferation. Proliferation-positive cells were scored and labeling index expressed as a percentage of positive nuclei from at least 3000 counted cells in mammary glands. A: Representative immunohistochemical

    Techniques Used: Labeling, Immunohistochemistry

    6) Product Images from "Whole ovary immunohistochemistry for monitoring cell proliferation and ovulatory wound repair in the mouse"

    Article Title: Whole ovary immunohistochemistry for monitoring cell proliferation and ovulatory wound repair in the mouse

    Journal: Reproductive Biology and Endocrinology : RB & E

    doi: 10.1186/1477-7827-8-98

    E-cadherin expression in the ovarian surface epithelial cells . Immunohistochemical staining for E-cadherin in a paraffin-embedded ovary section (A) and whole ovaries (B, C). The asterisks indicate groups of epithelial cells in which E-cadherin is expressed at a low level. Distinct patches of ovarian surface epithelial cells with variable levels of E-cadherin expression are detected by whole ovary immunohistochemistry (C).
    Figure Legend Snippet: E-cadherin expression in the ovarian surface epithelial cells . Immunohistochemical staining for E-cadherin in a paraffin-embedded ovary section (A) and whole ovaries (B, C). The asterisks indicate groups of epithelial cells in which E-cadherin is expressed at a low level. Distinct patches of ovarian surface epithelial cells with variable levels of E-cadherin expression are detected by whole ovary immunohistochemistry (C).

    Techniques Used: Expressing, Immunohistochemistry, Staining

    7) Product Images from "Periostin Secreted by Glioblastoma Stem Cells Recruits M2 Tumor-associated Macrophages and Promotes Malignant Growth"

    Article Title: Periostin Secreted by Glioblastoma Stem Cells Recruits M2 Tumor-associated Macrophages and Promotes Malignant Growth

    Journal: Nature cell biology

    doi: 10.1038/ncb3090

    TAMs in human primary GBMs and xenografts are monocyte-derived macrophages from peripheral blood a, Immunofluorescent staining of CCR2 (the marker for monocyte-derived macrophages) and CX3CR1 (the microglia marker) in GBM xenograft and normal brain tissue. Frozen sections of GSC-derived tumor (T387) and the adjacent normal mouse brain were immunostained with antibodies against CCR2 (green) and CX3CR1 (red) and counterstained with DAPI (blue). CCR2 + cells were detected only in tumor tissue, while CX3CR1 + cells were detected only in normal brain. Scale bar, 40μm. b , Immunohistochemical staining of CCR2 and CX3CR1 in human primary GBMs and normal brain tissue. Two consecutive tissue microarray slides (US Biomax) containing GBMs and normal brain tissues were immunostained with antibody against CCR2 or CX3CR1 (brown) and counterstained with hematoxylin. GBM tumors showed abundant CCR2 + cells (the monocyte-derived TAMs) but not CX3CR1 + cells (microglia), while normal brain tissues contained CX3CR1 + cells but not CCR2 + cells. Scale bar, 40μm. c , Graphical analysis of ( b ) in tissue microarrays showed that CCR2 + cells (monocyte-derived TAMs) were detected in the majority (82.6%) of GBM cases (data from 23 tumors) and the minority (20%) of normal brains (data from 5 normal samples). In contrast, CX3CR1 + cells (microglia) were detected in all normal brains but only 4.3% of GBM tumor cases. d , Immunofluorescent staining of Iba1 and the CD31 in human primary GBMs. Frozen sections of a primary GBM (CW2445) were immunostained with antibodies against Iba1 to label TAMs (green) and CD31 to mark vessels (red) and counterstained with DAPI (blue). Abundant TAMs are enriched in perivascular niches. Scale bar, 80μm. e , Immunofluorescent staining of the TAM marker Iba1 and the endothelial marker Glut1 in GSC-derived xenografts. Frozen sections of GBM xenografts derived from GSCs (T387) were immunostained with antibodies against Iba1 (green) and Glut1 (red) and counterstained with DAPI (blue). TAMs (green) were localized near vessels but not in the area lacking blood vessel. Scale bar, 80μm.
    Figure Legend Snippet: TAMs in human primary GBMs and xenografts are monocyte-derived macrophages from peripheral blood a, Immunofluorescent staining of CCR2 (the marker for monocyte-derived macrophages) and CX3CR1 (the microglia marker) in GBM xenograft and normal brain tissue. Frozen sections of GSC-derived tumor (T387) and the adjacent normal mouse brain were immunostained with antibodies against CCR2 (green) and CX3CR1 (red) and counterstained with DAPI (blue). CCR2 + cells were detected only in tumor tissue, while CX3CR1 + cells were detected only in normal brain. Scale bar, 40μm. b , Immunohistochemical staining of CCR2 and CX3CR1 in human primary GBMs and normal brain tissue. Two consecutive tissue microarray slides (US Biomax) containing GBMs and normal brain tissues were immunostained with antibody against CCR2 or CX3CR1 (brown) and counterstained with hematoxylin. GBM tumors showed abundant CCR2 + cells (the monocyte-derived TAMs) but not CX3CR1 + cells (microglia), while normal brain tissues contained CX3CR1 + cells but not CCR2 + cells. Scale bar, 40μm. c , Graphical analysis of ( b ) in tissue microarrays showed that CCR2 + cells (monocyte-derived TAMs) were detected in the majority (82.6%) of GBM cases (data from 23 tumors) and the minority (20%) of normal brains (data from 5 normal samples). In contrast, CX3CR1 + cells (microglia) were detected in all normal brains but only 4.3% of GBM tumor cases. d , Immunofluorescent staining of Iba1 and the CD31 in human primary GBMs. Frozen sections of a primary GBM (CW2445) were immunostained with antibodies against Iba1 to label TAMs (green) and CD31 to mark vessels (red) and counterstained with DAPI (blue). Abundant TAMs are enriched in perivascular niches. Scale bar, 80μm. e , Immunofluorescent staining of the TAM marker Iba1 and the endothelial marker Glut1 in GSC-derived xenografts. Frozen sections of GBM xenografts derived from GSCs (T387) were immunostained with antibodies against Iba1 (green) and Glut1 (red) and counterstained with DAPI (blue). TAMs (green) were localized near vessels but not in the area lacking blood vessel. Scale bar, 80μm.

    Techniques Used: Derivative Assay, Staining, Marker, Immunohistochemistry, Microarray

    POSTN is preferentially secreted by GSCs and its levels correlate with TAM density in GBMs a, Immunofluorescent staining of POSTN (green) and the GSC marker OLIG2 or SOX2 (red) in human primary GBMs. Frozen sections of GBMs (CCF2445 and CW672) were immunostained with antibodies against POSTN and SOX2 or OLIG2, and counterstained with DAPI to show nuclei (blue). POSTN was preferentially expressed in GSCs and distributed in the area near GSCs. Scale bar, 20μm. b , Graphical analysis of ( a ) showing the fraction of POSTN+ cells in SOX2+ cells (GSCs) in human primary GBMs. More than 70% of SOX2 positive GSCs showed POSTN staining (n=5 GBMs; mean ± s.e.m.). c and d , Immunoblot analysis of POSTN expression in cell lysates ( c ) and conditioned media (CM) ( d ) from GSCs (+) and matched non-stem tumor cells (−). CMs were obtained by culturing equal numbers of GSCs and non-stem tumor cells in Neurobasal media without supplements for 24 hours and concentrating media by vacuum centrifugation. Endogenous tubulin amounts in the corresponding cells were used for control. e, Immunofluorescent analysis of POSTN (red) and the TAM marker Iba1 (green) in GBM tissue microarray (US Biomax). Two sets of representative staining were presented to show the enrichment of TAMs in POSTN abundant regions in GBMs. Areas indicated with squares were enlarged and shown on right side of each picture. Scale bar, 80μm. f, Immunohistochemical staining of POSTN and the TAM marker Iba1 in two consecutive tissue microarray slides, respectively. Representative staining images show that the GBM (GL803a-C6) with higher POSTN levels contained more Iba1+ cells (TAMs) and the GBM (GL803a-E7) with lower POSTN levels has less Iba1+ (TAMs). Scale bar, 40μm. g , Graphical analysis of POSTN and Iba1 staining in the tissue microarray slides. 52.5% of GBM cases showed POSTN High and Iba1 High staining, and 28.75% of GBM cases showed POSTN Low and Iba1 Low staining. Only 12.5% of GBM cases showed POSTN High but Iba1 Low staining, and 6.25% of GBM cases showed POSTN Low but Iba1 High staining. The majority (81.25%) of GBM cases showed that POSTN levels positively correlate with TAM density. (Data from 80 tumors).
    Figure Legend Snippet: POSTN is preferentially secreted by GSCs and its levels correlate with TAM density in GBMs a, Immunofluorescent staining of POSTN (green) and the GSC marker OLIG2 or SOX2 (red) in human primary GBMs. Frozen sections of GBMs (CCF2445 and CW672) were immunostained with antibodies against POSTN and SOX2 or OLIG2, and counterstained with DAPI to show nuclei (blue). POSTN was preferentially expressed in GSCs and distributed in the area near GSCs. Scale bar, 20μm. b , Graphical analysis of ( a ) showing the fraction of POSTN+ cells in SOX2+ cells (GSCs) in human primary GBMs. More than 70% of SOX2 positive GSCs showed POSTN staining (n=5 GBMs; mean ± s.e.m.). c and d , Immunoblot analysis of POSTN expression in cell lysates ( c ) and conditioned media (CM) ( d ) from GSCs (+) and matched non-stem tumor cells (−). CMs were obtained by culturing equal numbers of GSCs and non-stem tumor cells in Neurobasal media without supplements for 24 hours and concentrating media by vacuum centrifugation. Endogenous tubulin amounts in the corresponding cells were used for control. e, Immunofluorescent analysis of POSTN (red) and the TAM marker Iba1 (green) in GBM tissue microarray (US Biomax). Two sets of representative staining were presented to show the enrichment of TAMs in POSTN abundant regions in GBMs. Areas indicated with squares were enlarged and shown on right side of each picture. Scale bar, 80μm. f, Immunohistochemical staining of POSTN and the TAM marker Iba1 in two consecutive tissue microarray slides, respectively. Representative staining images show that the GBM (GL803a-C6) with higher POSTN levels contained more Iba1+ cells (TAMs) and the GBM (GL803a-E7) with lower POSTN levels has less Iba1+ (TAMs). Scale bar, 40μm. g , Graphical analysis of POSTN and Iba1 staining in the tissue microarray slides. 52.5% of GBM cases showed POSTN High and Iba1 High staining, and 28.75% of GBM cases showed POSTN Low and Iba1 Low staining. Only 12.5% of GBM cases showed POSTN High but Iba1 Low staining, and 6.25% of GBM cases showed POSTN Low but Iba1 High staining. The majority (81.25%) of GBM cases showed that POSTN levels positively correlate with TAM density. (Data from 80 tumors).

    Techniques Used: Staining, Marker, Expressing, Centrifugation, Microarray, Immunohistochemistry

    8) Product Images from "Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model"

    Article Title: Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

    Journal: Canadian Journal of Veterinary Research

    doi:

    Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P
    Figure Legend Snippet: Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

    Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.
    Figure Legend Snippet: Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.

    Techniques Used: Western Blot, Recombinant, Produced

    9) Product Images from "The sinusoidal probe: a new approach to improve electrode longevity"

    Article Title: The sinusoidal probe: a new approach to improve electrode longevity

    Journal: Frontiers in Neuroengineering

    doi: 10.3389/fneng.2014.00010

    The neurofilament response over a 12 (rabbit p) and 24 (rabbit-N) month chronic indwelling period. (A) Representative images for the neurofilament response for both electrode types at the 6 month time point. (B) Representative images for the neurofilament response for both electrode types at the 12 month time point. The white dot represents the microwire electrode implantation site. (C) Overall normalized integrated intensity response (±SEM) for both electrode types for the 12 month time point ( n = 15 electrode tracts per electrode comparison). Significant differences were found at 50–500 μm away from the electrode implantation site ( P
    Figure Legend Snippet: The neurofilament response over a 12 (rabbit p) and 24 (rabbit-N) month chronic indwelling period. (A) Representative images for the neurofilament response for both electrode types at the 6 month time point. (B) Representative images for the neurofilament response for both electrode types at the 12 month time point. The white dot represents the microwire electrode implantation site. (C) Overall normalized integrated intensity response (±SEM) for both electrode types for the 12 month time point ( n = 15 electrode tracts per electrode comparison). Significant differences were found at 50–500 μm away from the electrode implantation site ( P

    Techniques Used:

    10) Product Images from "Structural analysis of a novel rabbit monoclonal antibody R53 targeting an epitope in HIV-1 gp120 C4 region critical for receptor and co-receptor binding"

    Article Title: Structural analysis of a novel rabbit monoclonal antibody R53 targeting an epitope in HIV-1 gp120 C4 region critical for receptor and co-receptor binding

    Journal: Emerging Microbes & Infections

    doi: 10.1038/emi.2015.44

    Structure of the Fab R53/epitope complex. ( A ) A ribbon representation of Fab R53 in complex with its epitope in a front view. The light chain, heavy chain and the R53 epitope are colored cyan, green, and magenta, respectively (a coloring scheme maintained throughout the manuscript, except where otherwise indicated). ( B ) A side view of the complex. ( C ) A top view of the R53/epitope complex looking at the antigen-binding site. The CDR regions are labeled and colored differently from the rest of the Fab. ( D ) Electrostatic surface potentials of R53, with red as the negatively charged and blue as positively charged regions. The inset is the sequence of the peptide used in crystallization, with the magenta region indicating residues visualized in the crystal structure.
    Figure Legend Snippet: Structure of the Fab R53/epitope complex. ( A ) A ribbon representation of Fab R53 in complex with its epitope in a front view. The light chain, heavy chain and the R53 epitope are colored cyan, green, and magenta, respectively (a coloring scheme maintained throughout the manuscript, except where otherwise indicated). ( B ) A side view of the complex. ( C ) A top view of the R53/epitope complex looking at the antigen-binding site. The CDR regions are labeled and colored differently from the rest of the Fab. ( D ) Electrostatic surface potentials of R53, with red as the negatively charged and blue as positively charged regions. The inset is the sequence of the peptide used in crystallization, with the magenta region indicating residues visualized in the crystal structure.

    Techniques Used: Binding Assay, Labeling, Sequencing, Crystallization Assay

    Details of the epitope binding in the Fab R53/epitope complex. ( A ) A stereo view of the antigen-binding site. The side chains of the key residues involved in hydrogen bonding and van der Waals interactions (with contact areas greater than 10 Å 2 ) are shown in sticks. Note that (i) three tyrosines and four negatively charged residues in R53 play a key role in binding the epitope, (ii) C4 residue Arg P440 interacts with two acidic residues Asp L30 and Glu L31 from the light chain, and (iii) the side chain of C4 residue Tyr P435 engages in van der Waals stacking with the antibody backbone, and its hydroxyl group forms a hydrogen bond with the side chain of Gl u H56 of the heavy chain. ( B ) A schematic of the antigen–antibody interaction. Hydrogen-binding interactions are indicated by dashed lines between the residues, whereas van der Waals contacts are indicated by eyelashes. Residues in solid ovals contribute to the interaction by their main chain atoms, and those in dashed ovals contribute to the interaction by their side chain atoms.
    Figure Legend Snippet: Details of the epitope binding in the Fab R53/epitope complex. ( A ) A stereo view of the antigen-binding site. The side chains of the key residues involved in hydrogen bonding and van der Waals interactions (with contact areas greater than 10 Å 2 ) are shown in sticks. Note that (i) three tyrosines and four negatively charged residues in R53 play a key role in binding the epitope, (ii) C4 residue Arg P440 interacts with two acidic residues Asp L30 and Glu L31 from the light chain, and (iii) the side chain of C4 residue Tyr P435 engages in van der Waals stacking with the antibody backbone, and its hydroxyl group forms a hydrogen bond with the side chain of Gl u H56 of the heavy chain. ( B ) A schematic of the antigen–antibody interaction. Hydrogen-binding interactions are indicated by dashed lines between the residues, whereas van der Waals contacts are indicated by eyelashes. Residues in solid ovals contribute to the interaction by their main chain atoms, and those in dashed ovals contribute to the interaction by their side chain atoms.

    Techniques Used: Binding Assay

    Locations and conformations of the R53 epitope region in gp120 structures. The structure of the R53 epitope, together with Fab R53 (only the Fv region is shown), was superimposed onto the epitope region of gp120 structures with distinct C4 conformations, including the CD4-bound ( A ), b12-bound ( B ), F105-bound ( C ), and b13-bound ( D ) gp120s. 11 , 25 , 26 , 27 The R53 epitope is colored magenta, while gp120 is colored gray. Clashes between R53 and gp120s are indicated. The R53 epitope is accessible (without clashes between the antibody and gp120) only in the b13-bound conformation of gp120. ( E ) The location of the R53 epitope in the recent published structure of BG505 SOSIP.664 trimer. The R53 epitope region, located underneath V1V2/V3, is colored magenta, while the rest of the three gp120s is colored gray, yellow, and orange, respectively.
    Figure Legend Snippet: Locations and conformations of the R53 epitope region in gp120 structures. The structure of the R53 epitope, together with Fab R53 (only the Fv region is shown), was superimposed onto the epitope region of gp120 structures with distinct C4 conformations, including the CD4-bound ( A ), b12-bound ( B ), F105-bound ( C ), and b13-bound ( D ) gp120s. 11 , 25 , 26 , 27 The R53 epitope is colored magenta, while gp120 is colored gray. Clashes between R53 and gp120s are indicated. The R53 epitope is accessible (without clashes between the antibody and gp120) only in the b13-bound conformation of gp120. ( E ) The location of the R53 epitope in the recent published structure of BG505 SOSIP.664 trimer. The R53 epitope region, located underneath V1V2/V3, is colored magenta, while the rest of the three gp120s is colored gray, yellow, and orange, respectively.

    Techniques Used:

    Broad reaction of R53 with gp120s of diverse clades. ( A ) Competition assay of R53 with Ig-CD4. The soluble CD4 (sCD4) and anti-V3 mAb R56 were used as positive and negative controls, respectively. Results shown represent the mean levels of two independent experiments and error bars indicate the standard deviations. ( B ) R53 bound nine gp120s from clades A, B, C, D, and AE, as revealed by ELISA. ( C ) Western blot analysis showed that five selected gp120 proteins from different clades were recognized by R53. ( D ) A protein sequence alignment of R53 epitope region of the gp120s used in panel B and C. The R53 epitope is underlined in the JR-FL sequence.
    Figure Legend Snippet: Broad reaction of R53 with gp120s of diverse clades. ( A ) Competition assay of R53 with Ig-CD4. The soluble CD4 (sCD4) and anti-V3 mAb R56 were used as positive and negative controls, respectively. Results shown represent the mean levels of two independent experiments and error bars indicate the standard deviations. ( B ) R53 bound nine gp120s from clades A, B, C, D, and AE, as revealed by ELISA. ( C ) Western blot analysis showed that five selected gp120 proteins from different clades were recognized by R53. ( D ) A protein sequence alignment of R53 epitope region of the gp120s used in panel B and C. The R53 epitope is underlined in the JR-FL sequence.

    Techniques Used: Competitive Binding Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Sequencing

    Binding kinetics of R53 and gp120s. The binding kinetics of R53 to 92UG037 (clade A), JR-FL (clade B), 93MW965 (clade C), 92UG021 (clade D), and AE consensus (clade AE) were measured using ForteBio at different concentrations of the ligand. The brown, red, blue, purple, green, and orange lines represent 300 nM, 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, and 1.23 nM of gp120 proteins, respectively. Dashed gray lines represent the theoretical fitting curves. The binding data are summarized in Table 2 .
    Figure Legend Snippet: Binding kinetics of R53 and gp120s. The binding kinetics of R53 to 92UG037 (clade A), JR-FL (clade B), 93MW965 (clade C), 92UG021 (clade D), and AE consensus (clade AE) were measured using ForteBio at different concentrations of the ligand. The brown, red, blue, purple, green, and orange lines represent 300 nM, 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, and 1.23 nM of gp120 proteins, respectively. Dashed gray lines represent the theoretical fitting curves. The binding data are summarized in Table 2 .

    Techniques Used: Binding Assay

    11) Product Images from "Affimer proteins are versatile and renewable affinity reagents"

    Article Title: Affimer proteins are versatile and renewable affinity reagents

    Journal: eLife

    doi: 10.7554/eLife.24903

    Western blot results of Avi-Tag SH2 domain proteins using an streptavidin-HRP conjugate to detect the presence of biotin. DOI: http://dx.doi.org/10.7554/eLife.24903.005
    Figure Legend Snippet: Western blot results of Avi-Tag SH2 domain proteins using an streptavidin-HRP conjugate to detect the presence of biotin. DOI: http://dx.doi.org/10.7554/eLife.24903.005

    Techniques Used: Western Blot

    12) Product Images from "MEK1 expression and its relationship with clinical pathological features in hepatocellular carcinoma"

    Article Title: MEK1 expression and its relationship with clinical pathological features in hepatocellular carcinoma

    Journal: International Journal of Clinical and Experimental Medicine

    doi:

    PCR detection and immunohistochemical staining for MEK1 expression in HCC cell lines. The expression of MEK1 mRNA (A) and protein (B, C) in three liver cell lines (Hun7, HepG3B and 7402) were significantly higher than controls (** P
    Figure Legend Snippet: PCR detection and immunohistochemical staining for MEK1 expression in HCC cell lines. The expression of MEK1 mRNA (A) and protein (B, C) in three liver cell lines (Hun7, HepG3B and 7402) were significantly higher than controls (** P

    Techniques Used: Polymerase Chain Reaction, Immunohistochemistry, Staining, Expressing

    13) Product Images from "Toll-like receptors-2, -3 and -4 expression patterns on human colon and their regulation by mucosal-associated bacteria"

    Article Title: Toll-like receptors-2, -3 and -4 expression patterns on human colon and their regulation by mucosal-associated bacteria

    Journal: Immunology

    doi: 10.1111/j.1365-2567.2005.02200.x

    Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse anti-human TLR2 IgG2a antibody, visualized using immunoperoxidase. (a) Transverse section (×40) across the crypt region with staining in the epithelium
    Figure Legend Snippet: Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse anti-human TLR2 IgG2a antibody, visualized using immunoperoxidase. (a) Transverse section (×40) across the crypt region with staining in the epithelium

    Techniques Used: Immunohistochemistry, Staining

    Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse anti-human TLR4 IgG2a antibody, visualized using immunoperoxidase. Expression patterns of human TLR4. (a)A transverse section (×40) across the crypt region
    Figure Legend Snippet: Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse anti-human TLR4 IgG2a antibody, visualized using immunoperoxidase. Expression patterns of human TLR4. (a)A transverse section (×40) across the crypt region

    Techniques Used: Immunohistochemistry, Staining, Expressing

    Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse antihuman TLR3 IgG1 antibody, visualized using immunoperoxidase. (a and b) Longitudinal sections (×40) through the epithelium at the luminal surface. (c)
    Figure Legend Snippet: Immunohistochemical analysis of normal healthy human colon stained with a monoclonal mouse antihuman TLR3 IgG1 antibody, visualized using immunoperoxidase. (a and b) Longitudinal sections (×40) through the epithelium at the luminal surface. (c)

    Techniques Used: Immunohistochemistry, Staining

    14) Product Images from "Deletion of Metallothionein Exacerbates Intermittent Hypoxia-Induced Oxidative and Inflammatory Injury in Aorta"

    Article Title: Deletion of Metallothionein Exacerbates Intermittent Hypoxia-Induced Oxidative and Inflammatory Injury in Aorta

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2014/141053

    MT-KO mice exhibited earlier and more severe IH-induced aortic oxidative damage. Aortic oxidative damage was examined by immunohistochemical staining for the accumulation of 3-NT (a) and 4-HNE (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P
    Figure Legend Snippet: MT-KO mice exhibited earlier and more severe IH-induced aortic oxidative damage. Aortic oxidative damage was examined by immunohistochemical staining for the accumulation of 3-NT (a) and 4-HNE (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining

    Effects of MT on eNOS and p47phox expression in aorta. Aortic eNOS (a) and p47phox (b) expression were examined by immunohistochemical staining, followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P
    Figure Legend Snippet: Effects of MT on eNOS and p47phox expression in aorta. Aortic eNOS (a) and p47phox (b) expression were examined by immunohistochemical staining, followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P

    Techniques Used: Expressing, Immunohistochemistry, Staining

    MT-KO mice exhibited earlier and more severe IH-induced aortic fibrosis. Aortic fibrosis was examined by immunohistochemical staining and qRT-PCR for the expression of CTGF (a) and immunohistochemical staining for TGF- β 1 (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P
    Figure Legend Snippet: MT-KO mice exhibited earlier and more severe IH-induced aortic fibrosis. Aortic fibrosis was examined by immunohistochemical staining and qRT-PCR for the expression of CTGF (a) and immunohistochemical staining for TGF- β 1 (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining, Quantitative RT-PCR, Expressing

    Effects of IH on aortic expression of MT. Aortic expression of MT was examined by immunohistochemical staining (a) with semiquantitative analysis (b). Data were presented as means ± SDs ( n = 6). * P
    Figure Legend Snippet: Effects of IH on aortic expression of MT. Aortic expression of MT was examined by immunohistochemical staining (a) with semiquantitative analysis (b). Data were presented as means ± SDs ( n = 6). * P

    Techniques Used: Expressing, Immunohistochemistry, Staining

    MT-KO mice exhibited earlier and more severe IH-induced aortic inflammation. Aortic inflammation was examined by immunohistochemical staining and qRT-PCR for the expression of TNF- α (a) and immunohistochemical staining for VCAM-1 (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P
    Figure Legend Snippet: MT-KO mice exhibited earlier and more severe IH-induced aortic inflammation. Aortic inflammation was examined by immunohistochemical staining and qRT-PCR for the expression of TNF- α (a) and immunohistochemical staining for VCAM-1 (b), followed by semiquantitative analysis. Data were presented as means ± SDs ( n = 6); * P

    Techniques Used: Mouse Assay, Immunohistochemistry, Staining, Quantitative RT-PCR, Expressing

    15) Product Images from "Inhibition of JNK by compound C66 prevents pathological changes of the aorta in STZ-induced diabetes"

    Article Title: Inhibition of JNK by compound C66 prevents pathological changes of the aorta in STZ-induced diabetes

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.12267

    Effects of C66 on aortic Nrf2 expression. The expression of Nrf2 was examined by immunohistochemical staining for Nrf2 (A) expression at protein level in aortic tunica media, followed semi-quantitative analysis (B) and real-time PCR for its expression at mRNA (C) level. Data were presented as means ± SD ( n = 5); * P
    Figure Legend Snippet: Effects of C66 on aortic Nrf2 expression. The expression of Nrf2 was examined by immunohistochemical staining for Nrf2 (A) expression at protein level in aortic tunica media, followed semi-quantitative analysis (B) and real-time PCR for its expression at mRNA (C) level. Data were presented as means ± SD ( n = 5); * P

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

    Effects of C66 on diabetes-induced mast cells evaluated around the aortic and oxidative stress damage. The mast cells around the aortic were identified by toluidine blue staining (A). Magnification at 400 × followed semi-quantitative analysis. The oxidative stress damage was examined by immunohistochemical staining for the accumulation of 3-NT (B), followed with semi-quantitative analysis. Data were presented as means ± SD ( n = 5); * P
    Figure Legend Snippet: Effects of C66 on diabetes-induced mast cells evaluated around the aortic and oxidative stress damage. The mast cells around the aortic were identified by toluidine blue staining (A). Magnification at 400 × followed semi-quantitative analysis. The oxidative stress damage was examined by immunohistochemical staining for the accumulation of 3-NT (B), followed with semi-quantitative analysis. Data were presented as means ± SD ( n = 5); * P

    Techniques Used: Staining, Immunohistochemistry

    Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and NQO-1 (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P
    Figure Legend Snippet: Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and NQO-1 (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P

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

    Protective effect of C66 on diabetes-induced aortic tumour necrosis factor-alpha (TNF-α) expression. Aortic expression of TNF-α was examined by immunohistochemical staining for its protein (A) expression in aortic tunica media, followed by semi-quantitative analysis (B) and real-time PCR for its mRNA level (C). Data were presented as means ± SD ( n = 5). * P
    Figure Legend Snippet: Protective effect of C66 on diabetes-induced aortic tumour necrosis factor-alpha (TNF-α) expression. Aortic expression of TNF-α was examined by immunohistochemical staining for its protein (A) expression in aortic tunica media, followed by semi-quantitative analysis (B) and real-time PCR for its mRNA level (C). Data were presented as means ± SD ( n = 5). * P

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

    Protective effect of C66 on diabetes-induced aortic inflammation. The inflammation markers PAI-1 was examined by immunohistochemical staining, followed by semi-quantitative analysis. Data were presented as means ± SD ( n = 5); * P
    Figure Legend Snippet: Protective effect of C66 on diabetes-induced aortic inflammation. The inflammation markers PAI-1 was examined by immunohistochemical staining, followed by semi-quantitative analysis. Data were presented as means ± SD ( n = 5); * P

    Techniques Used: Immunohistochemistry, Staining

    16) Product Images from "Caspase-8 Collaborates with Caspase-11 to Drive Tissue Damage and Execution of Endotoxic Shock"

    Article Title: Caspase-8 Collaborates with Caspase-11 to Drive Tissue Damage and Execution of Endotoxic Shock

    Journal: Immunity

    doi: 10.1016/j.immuni.2018.06.011

    Casp8 regulates inflammatory signaling in ileum (A) Immunoblot of ileal mucosa from unmanipulated (time=0) or from WT and Casp8 −/− Ripk3 −/− mice at the indicated mpc with LPS, showing expression of phosphorylated forms (p) of IκBα (40 kDa), SAPK and JNK (54 and 46 kDa, respectively), as well as Cl-C8, C8, cFLIP L (50 kDa) and cFLIP s (25 kDa), RIPK1 (75 kDa), IgH (50 kDa), C3, C11, C1 (43 kDa) and Gsdmd (55 kDa, with an * indicating the ~30 kDa cleaved form). (B) Principal component (PC) analysis (PCA) of ileal mucosa from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice (n=5 for each group) including 27,578 genes with detectable base mean expression where the contribution of each PC is indicated as a percentage. (C) Heat map comparing expression patterns of genes in ileal mucosa from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice (n=5 for each group) at 1.5 hpc, where mean log 2 .
    Figure Legend Snippet: Casp8 regulates inflammatory signaling in ileum (A) Immunoblot of ileal mucosa from unmanipulated (time=0) or from WT and Casp8 −/− Ripk3 −/− mice at the indicated mpc with LPS, showing expression of phosphorylated forms (p) of IκBα (40 kDa), SAPK and JNK (54 and 46 kDa, respectively), as well as Cl-C8, C8, cFLIP L (50 kDa) and cFLIP s (25 kDa), RIPK1 (75 kDa), IgH (50 kDa), C3, C11, C1 (43 kDa) and Gsdmd (55 kDa, with an * indicating the ~30 kDa cleaved form). (B) Principal component (PC) analysis (PCA) of ileal mucosa from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice (n=5 for each group) including 27,578 genes with detectable base mean expression where the contribution of each PC is indicated as a percentage. (C) Heat map comparing expression patterns of genes in ileal mucosa from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice (n=5 for each group) at 1.5 hpc, where mean log 2 .

    Techniques Used: Mouse Assay, Expressing

    Casp8 and Casp11 mediate gut leakage (A) IHC and H E of ileum from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice at 6 hpc with LPS (bar=100 μm). Arrows indicate Cl-C3 positive cells. (B) IHC of ileum from Irf3 −/− mice or Irf3 −/− recipients of BMC from WT (n=3 each) mice at 6 to 9 hpc with LPS. (C) Detection of FD-4 in sera from WT (n=10), Ripk3 −/− (n=10), Casp8 −/− Ripk3 −/− (n=12), Ifnar1 −/− (n=6) and Casp11 −/− (n=6) mice at 6 hpc. (Statistical comparisons used Wilcoxan matched pairs analysis (* p
    Figure Legend Snippet: Casp8 and Casp11 mediate gut leakage (A) IHC and H E of ileum from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice at 6 hpc with LPS (bar=100 μm). Arrows indicate Cl-C3 positive cells. (B) IHC of ileum from Irf3 −/− mice or Irf3 −/− recipients of BMC from WT (n=3 each) mice at 6 to 9 hpc with LPS. (C) Detection of FD-4 in sera from WT (n=10), Ripk3 −/− (n=10), Casp8 −/− Ripk3 −/− (n=12), Ifnar1 −/− (n=6) and Casp11 −/− (n=6) mice at 6 hpc. (Statistical comparisons used Wilcoxan matched pairs analysis (* p

    Techniques Used: Immunohistochemistry, Mouse Assay

    Casp8 drives small intestinal and splenic damage (A–B) Representative (3 mice of each genotype in two independent experiments) images of hematoxylin and eosin (H E) stained sections of small intestine (A) and spleen (B) , from WT, Ripk3 −/− and Casp8 −/− Ripk3 −/− mice at 9 hpc with PBS or LPS, as indicated (bar=100 μm). Arrows indicate damaged and sloughed cells with condensed nuclei. (C–D) Histological score of leukocyte infiltrate (inflammatory cells), condensed nuclei (apoptosis), atrophy (tissue damage) and edema (fluid accumulation) in small intestine (C) , or extramedullary hematopoiesis (EMH) as well as congestion in spleen (D), in tissue sections from WT, Ripk3 −/− and Casp8 −/− Ripk3 −/− .
    Figure Legend Snippet: Casp8 drives small intestinal and splenic damage (A–B) Representative (3 mice of each genotype in two independent experiments) images of hematoxylin and eosin (H E) stained sections of small intestine (A) and spleen (B) , from WT, Ripk3 −/− and Casp8 −/− Ripk3 −/− mice at 9 hpc with PBS or LPS, as indicated (bar=100 μm). Arrows indicate damaged and sloughed cells with condensed nuclei. (C–D) Histological score of leukocyte infiltrate (inflammatory cells), condensed nuclei (apoptosis), atrophy (tissue damage) and edema (fluid accumulation) in small intestine (C) , or extramedullary hematopoiesis (EMH) as well as congestion in spleen (D), in tissue sections from WT, Ripk3 −/− and Casp8 −/− Ripk3 −/− .

    Techniques Used: Mouse Assay, Staining

    Casp8 and Casp11 collaborate for lower small intestinal injury (A–C) Immunoblot for cleaved (Cl)-Casp (C)8; 43 and 18 kDa), Cl-C3 (22, 19 and 17 kDa) or Cl-PARP (89 kDa) as well as total C11 (43, 38.5 and 25 kDa) and uncleaved C8 (55 kDa) or C3 (31 kDa), showing mucosa from duodenum (Du) in challenged WT and Casp8 −/− Ripk3 −/− mice (only uncleaved 43 and 38.5 kDa forms of C11 detected) (A) , PBMC and Du from WT mice (B) , mucosa from Du, as well as stomach (S), proximal jejunum (Jp), distal jejunum (Jd), ileum (Ile), cecum (Ce) and colon (Co) from challenged WT mice (only 19 and 17 kDa forms of Cl-C3 detected) (C) . Graphical depiction of intestinal tract segments is above C. Molecular weight markers shown to the right. (D) Immunohistochemistry (IHC), showing Cl-C3 in situ with hematoxylin counterstain, or histology following H E stain of the indicated intestinal segments from challenged WT and Casp8 −/− Ripk3 −/− mice (bar=100 μm) representative of 8 mice from 4 independent experiments. Insets show magnified images of the sections (bar=40 μm). Arrows indicate Cl-C3 positive cells in IHC sections and condensed nuclei in H E stained sections. (E) Immunoblot of mucosa from Du, Jp, Jd and Ile from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice at 1.5 hpc with LPS. (F) IHC of Cl-C3 in ileum from two WT and two Casp11 −/− mice at 1.5 hpc with PBS or LPS representative of 5 mice from 3 independent experiments. Arrows indicate Cl-C3 positive cells. (G) Immunoblot of ileal mucosa from WT and Irf3 −/− mice at 1.5 hpc with PBS or LPS, with two LPS-challenged Irf3 −/− .
    Figure Legend Snippet: Casp8 and Casp11 collaborate for lower small intestinal injury (A–C) Immunoblot for cleaved (Cl)-Casp (C)8; 43 and 18 kDa), Cl-C3 (22, 19 and 17 kDa) or Cl-PARP (89 kDa) as well as total C11 (43, 38.5 and 25 kDa) and uncleaved C8 (55 kDa) or C3 (31 kDa), showing mucosa from duodenum (Du) in challenged WT and Casp8 −/− Ripk3 −/− mice (only uncleaved 43 and 38.5 kDa forms of C11 detected) (A) , PBMC and Du from WT mice (B) , mucosa from Du, as well as stomach (S), proximal jejunum (Jp), distal jejunum (Jd), ileum (Ile), cecum (Ce) and colon (Co) from challenged WT mice (only 19 and 17 kDa forms of Cl-C3 detected) (C) . Graphical depiction of intestinal tract segments is above C. Molecular weight markers shown to the right. (D) Immunohistochemistry (IHC), showing Cl-C3 in situ with hematoxylin counterstain, or histology following H E stain of the indicated intestinal segments from challenged WT and Casp8 −/− Ripk3 −/− mice (bar=100 μm) representative of 8 mice from 4 independent experiments. Insets show magnified images of the sections (bar=40 μm). Arrows indicate Cl-C3 positive cells in IHC sections and condensed nuclei in H E stained sections. (E) Immunoblot of mucosa from Du, Jp, Jd and Ile from WT, Casp8 −/− Ripk3 −/− and Casp11 −/− mice at 1.5 hpc with LPS. (F) IHC of Cl-C3 in ileum from two WT and two Casp11 −/− mice at 1.5 hpc with PBS or LPS representative of 5 mice from 3 independent experiments. Arrows indicate Cl-C3 positive cells. (G) Immunoblot of ileal mucosa from WT and Irf3 −/− mice at 1.5 hpc with PBS or LPS, with two LPS-challenged Irf3 −/− .

    Techniques Used: Mouse Assay, Molecular Weight, Immunohistochemistry, In Situ, Staining

    Casp8 drives endotoxic shock (A–B) Kaplan-Meier survival plot (left) and body temperature plot (right) over time (hpc) in LPS-challenged WT (n=19), Ripk3 −/− (n=15), Casp8 +/− Ripk3 −/− (n=8), Casp8 −/− Ripk3 −/− (n=26) and Casp11 −/− (n=11) mice (A) or WT (n=6), Ifnar1 −/− (n=8) and Irf3 −/− (n=5) mice (B). (C) Kaplan-Meier survival plot for WT (n=6), littermate Casp8 +/− Ripk3 −/− (n=6) and Casp8 −/− Ripk3 −/− (n=7) as well as Casp11 −/− mice (n=6) given a poly(I:C) prime (4 mg/kg) followed 6 h later with low dose LPS challenge (5 mg/kg). (D) Kaplan-Meier survival plot for WT (n=19), littermate Casp8 +/− Ripk3 −/− (n=20) and Casp8 −/− Ripk3 −/− (n=15) mice over time, h post infection (hpi) with E. coli .
    Figure Legend Snippet: Casp8 drives endotoxic shock (A–B) Kaplan-Meier survival plot (left) and body temperature plot (right) over time (hpc) in LPS-challenged WT (n=19), Ripk3 −/− (n=15), Casp8 +/− Ripk3 −/− (n=8), Casp8 −/− Ripk3 −/− (n=26) and Casp11 −/− (n=11) mice (A) or WT (n=6), Ifnar1 −/− (n=8) and Irf3 −/− (n=5) mice (B). (C) Kaplan-Meier survival plot for WT (n=6), littermate Casp8 +/− Ripk3 −/− (n=6) and Casp8 −/− Ripk3 −/− (n=7) as well as Casp11 −/− mice (n=6) given a poly(I:C) prime (4 mg/kg) followed 6 h later with low dose LPS challenge (5 mg/kg). (D) Kaplan-Meier survival plot for WT (n=19), littermate Casp8 +/− Ripk3 −/− (n=20) and Casp8 −/− Ripk3 −/− (n=15) mice over time, h post infection (hpi) with E. coli .

    Techniques Used: Mouse Assay, Infection

    17) Product Images from "Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model"

    Article Title: Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

    Journal: Canadian Journal of Veterinary Research

    doi:

    Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P
    Figure Legend Snippet: Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

    Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.
    Figure Legend Snippet: Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.

    Techniques Used: Western Blot, Recombinant, Produced

    18) Product Images from "Estrogen reprograms the activity of neutrophils to foster protumoral microenvironment during mammary involution"

    Article Title: Estrogen reprograms the activity of neutrophils to foster protumoral microenvironment during mammary involution

    Journal: Scientific Reports

    doi: 10.1038/srep46485

    Estrogen promotes neutrophil recruitment during mammary tissue involution. Mice at 24 h post-weaning or age-matched nulliparous mice were treated with Ctrl or E2B in sesame oil for 48 h. Mammary tissues were digested by collagenase to obtain single cell suspension. Mammary cells and blood samples were stained with various cell surface markers.( a ) E2B treatment did not affect total CD45 + cell infiltration in involuting or nulliparous mammary tissue. Representative histograms were shown for CD45 + in Ctrl- and E2B-treated nulliparous and involuting mammary glands.( b ) Estrogen treatment induced significant neutrophils and myeloid-derived monocytic cells infiltration to involuting but not to nulliparous mammary tissue.(Left) Representative dot plots of neutrophils(CD45 + CD11b + Gr-1 hi )(red boxes) and myeloid-derived monocytic cells(CD45 + CD11b + Gr1 int )(blue boxes) from mammary glands CD45+ cell population.(Right) Bar graphs show the percentages of neutrophils and myeloid-derived monocytic cells in total cell population. For the data on mammary tissue in( a , b ) from nulliparous mice, Ctrl n = 10, E2B n = 9; for the data of mammary tissue in( a , b ) from mice undergoing involution, Ctrl n = 13, E2B n = 13.( c ) Gr-1 IHC of Ctrl- or E2B-treated involuting mammary tissue. Scale bar = 50 μm.( d ) Total number of infiltrated neutrophils in mammary tissue were counted in an area of 20 mm 2 for each sample. n = 9 per group. White and black bars represent Ctrl and E2B treatment, respectively. Statistical significance was evaluated by unpaired two-tailed Student’s t-tests, **p
    Figure Legend Snippet: Estrogen promotes neutrophil recruitment during mammary tissue involution. Mice at 24 h post-weaning or age-matched nulliparous mice were treated with Ctrl or E2B in sesame oil for 48 h. Mammary tissues were digested by collagenase to obtain single cell suspension. Mammary cells and blood samples were stained with various cell surface markers.( a ) E2B treatment did not affect total CD45 + cell infiltration in involuting or nulliparous mammary tissue. Representative histograms were shown for CD45 + in Ctrl- and E2B-treated nulliparous and involuting mammary glands.( b ) Estrogen treatment induced significant neutrophils and myeloid-derived monocytic cells infiltration to involuting but not to nulliparous mammary tissue.(Left) Representative dot plots of neutrophils(CD45 + CD11b + Gr-1 hi )(red boxes) and myeloid-derived monocytic cells(CD45 + CD11b + Gr1 int )(blue boxes) from mammary glands CD45+ cell population.(Right) Bar graphs show the percentages of neutrophils and myeloid-derived monocytic cells in total cell population. For the data on mammary tissue in( a , b ) from nulliparous mice, Ctrl n = 10, E2B n = 9; for the data of mammary tissue in( a , b ) from mice undergoing involution, Ctrl n = 13, E2B n = 13.( c ) Gr-1 IHC of Ctrl- or E2B-treated involuting mammary tissue. Scale bar = 50 μm.( d ) Total number of infiltrated neutrophils in mammary tissue were counted in an area of 20 mm 2 for each sample. n = 9 per group. White and black bars represent Ctrl and E2B treatment, respectively. Statistical significance was evaluated by unpaired two-tailed Student’s t-tests, **p

    Techniques Used: Mouse Assay, Staining, Derivative Assay, Immunohistochemistry, Two Tailed Test

    19) Product Images from "Pharmacokinetics and Safety in Rhesus Monkeys of a Monoclonal Antibody-GDNF Fusion Protein for Targeted Blood-Brain Barrier Delivery"

    Article Title: Pharmacokinetics and Safety in Rhesus Monkeys of a Monoclonal Antibody-GDNF Fusion Protein for Targeted Blood-Brain Barrier Delivery

    Journal: Pharmaceutical Research

    doi: 10.1007/s11095-009-9939-6

    ( A ) Structure of the 2-site ELISA for detection of antibodies against the HIRMAb-GDNF fusion protein. The HIRMAb or the HIRMAb-GDNF fusion protein is used as the capture reagent, and the biotinylated HIRMAb-GDNF fusion protein is used as the detector reagent, along with a complex of streptavidin (SA) and horseradish peroxidase (HRP); the biotin moiety is designated, ‘B’. ( B ) Absorbance at various dilutions of a rabbit antiserum prepared against the HIRMAb-GDNF fusion protein, and the respective pre-immune serum (triangles). Either the HIRMAb (squares) or the HIRMAb-GDNF fusion protein (diamonds) was used as the capture reagent. Data are means of duplicates that varied
    Figure Legend Snippet: ( A ) Structure of the 2-site ELISA for detection of antibodies against the HIRMAb-GDNF fusion protein. The HIRMAb or the HIRMAb-GDNF fusion protein is used as the capture reagent, and the biotinylated HIRMAb-GDNF fusion protein is used as the detector reagent, along with a complex of streptavidin (SA) and horseradish peroxidase (HRP); the biotin moiety is designated, ‘B’. ( B ) Absorbance at various dilutions of a rabbit antiserum prepared against the HIRMAb-GDNF fusion protein, and the respective pre-immune serum (triangles). Either the HIRMAb (squares) or the HIRMAb-GDNF fusion protein (diamonds) was used as the capture reagent. Data are means of duplicates that varied

    Techniques Used: Enzyme-linked Immunosorbent Assay

    20) Product Images from "Connexin36 Is Essential for Transmission of Rod-Mediated Visual Signals in the Mammalian Retina"

    Article Title: Connexin36 Is Essential for Transmission of Rod-Mediated Visual Signals in the Mammalian Retina

    Journal: Neuron

    doi:

    β-Gal Reporter Is Present in the ONL, Amacrine, and Bipolar Cell Layers of the INL and Some Small Cells in the GCL (A) β-gal immunofluorescence visualized by confocal microscopy can be detected throughout the ONL, and the reporter distinctively labels the photoreceptor inner segments (asterisk). Furthermore, in the INL, β-gal labels cells with characteristic bipolar cell morphology (examples indicated with arrows) and amacrine cells immediately adjacent to the IPL (examples indicated with arrowheads). (B) β-gal immunohistochemistry detects the reporter in a limited number of neurons with small cell bodies in the GCL that may be displaced amacrine cells (arrow), and also in processes from the IPL immediately adjacent to the GCL (arrowhead). (C) No reporter can be detected by immunohistochemistry in WT retina. Scale bar equals 10 μm.
    Figure Legend Snippet: β-Gal Reporter Is Present in the ONL, Amacrine, and Bipolar Cell Layers of the INL and Some Small Cells in the GCL (A) β-gal immunofluorescence visualized by confocal microscopy can be detected throughout the ONL, and the reporter distinctively labels the photoreceptor inner segments (asterisk). Furthermore, in the INL, β-gal labels cells with characteristic bipolar cell morphology (examples indicated with arrows) and amacrine cells immediately adjacent to the IPL (examples indicated with arrowheads). (B) β-gal immunohistochemistry detects the reporter in a limited number of neurons with small cell bodies in the GCL that may be displaced amacrine cells (arrow), and also in processes from the IPL immediately adjacent to the GCL (arrowhead). (C) No reporter can be detected by immunohistochemistry in WT retina. Scale bar equals 10 μm.

    Techniques Used: Immunofluorescence, Confocal Microscopy, Immunohistochemistry

    21) Product Images from "p38-MAPK/MSK1-mediated overexpression of histone H3 serine 10 phosphorylation defines distance-dependent prognostic value of negative resection margin in gastric cancer"

    Article Title: p38-MAPK/MSK1-mediated overexpression of histone H3 serine 10 phosphorylation defines distance-dependent prognostic value of negative resection margin in gastric cancer

    Journal: Clinical Epigenetics

    doi: 10.1186/s13148-016-0255-9

    H3S10ph level in paired tumors and negative PRM and DRM GC tissue samples: a immunoblot analysis of H3S10ph, H4K16ac, and H4K20me3 in freshly resected paired tumor and resection margin tissues ( n = 10). b Representative image ( left panel , ×40) and comparison of mean H-score ( right panel ) for H4K16ac and H4K20me3 immunostaining ( n = 10). c Representative images (×40) of H3S10ph immunostaining ( n = 101). d Comparison of frequency distribution with low (H-score 0–100), intermediate (H-score 100–200), and high (H-score 200–300) levels of H3S10ph immunostaining ( n = 101). e Comparative mean H-score of H3S10ph immunostaining ( n = 101). GC gastric cancer, PRM proximal resection margin, DRM distal resection margin, RM resection margin either PRM or DRM with maximum distance from the site of the tumor, P patient. Statistical tests are done by using Wilcoxon matched pairs test. * p
    Figure Legend Snippet: H3S10ph level in paired tumors and negative PRM and DRM GC tissue samples: a immunoblot analysis of H3S10ph, H4K16ac, and H4K20me3 in freshly resected paired tumor and resection margin tissues ( n = 10). b Representative image ( left panel , ×40) and comparison of mean H-score ( right panel ) for H4K16ac and H4K20me3 immunostaining ( n = 10). c Representative images (×40) of H3S10ph immunostaining ( n = 101). d Comparison of frequency distribution with low (H-score 0–100), intermediate (H-score 100–200), and high (H-score 200–300) levels of H3S10ph immunostaining ( n = 101). e Comparative mean H-score of H3S10ph immunostaining ( n = 101). GC gastric cancer, PRM proximal resection margin, DRM distal resection margin, RM resection margin either PRM or DRM with maximum distance from the site of the tumor, P patient. Statistical tests are done by using Wilcoxon matched pairs test. * p

    Techniques Used: Immunostaining

    Association of H3S10ph with the distance of negative resection margins in GC. a , c Resection margins were grouped as per their distance from the site of the tumor with 1 cm interval and mean H-score of H3S10ph immunostaining of each group was compared with tumor ( left panel ). In case of both PRM and DRM, analysis showed a significant decrease in H3S10ph levels as the margin length reaches more than 4 cm ( left panel ). Comparison of mean H-score of H3S10ph immunostaining of all resection margins with a margin distance ≤4 and > 4 cm with tumor confirms the significant reduction of H3S10ph if the margin distance is > 4 cm ( right panel ). b , d Confirmation of reduction of H3S10ph, if the margin length is > 4 cm by immunoblotting. GC gastric cancer, PRM proximal resection margin, DRM distal resection margin. Statistical tests are done by using Mann-Whitney test ( † ) and Wilcoxon matched pairs test. * p
    Figure Legend Snippet: Association of H3S10ph with the distance of negative resection margins in GC. a , c Resection margins were grouped as per their distance from the site of the tumor with 1 cm interval and mean H-score of H3S10ph immunostaining of each group was compared with tumor ( left panel ). In case of both PRM and DRM, analysis showed a significant decrease in H3S10ph levels as the margin length reaches more than 4 cm ( left panel ). Comparison of mean H-score of H3S10ph immunostaining of all resection margins with a margin distance ≤4 and > 4 cm with tumor confirms the significant reduction of H3S10ph if the margin distance is > 4 cm ( right panel ). b , d Confirmation of reduction of H3S10ph, if the margin length is > 4 cm by immunoblotting. GC gastric cancer, PRM proximal resection margin, DRM distal resection margin. Statistical tests are done by using Mann-Whitney test ( † ) and Wilcoxon matched pairs test. * p

    Techniques Used: Immunostaining, MANN-WHITNEY

    22) Product Images from "CD147 and Cyclooxygenase Expression in Feline Oral Squamous Cell Carcinoma"

    Article Title: CD147 and Cyclooxygenase Expression in Feline Oral Squamous Cell Carcinoma

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5030072

    COX-2 and CD147 positive and negative controls for immunohistochemistry. ( A – D ) Photomicrographs of IHC staining using rabbit anti-COX-2 IgG (1:200) and normal rabbit IgG (1:200, negative control). The chromogen is DAB (brown), and the counterstain is hematoxylin (blue). The feline renal macula densa epithelial cells had an intense cytoplasmic signal ( A ) that was eliminated when the antibody was replaced with normal IgG ( B ). Scattered oral squamous cell carcinoma (OSCC) cells showed an intense cytoplasmic COX-2 signal ( C ), which was eliminated when the primary antibody was replaced with normal IgG ( D ). ( E , F ) IHC staining using goat anti-CD147 IgG (1:100) and normal goat IgG (1:100, negative control). Feline enterocytes showed moderately intense cytoplasmic and membranous signal ( E ), which was markedly reduced when the primary antibody was replaced with normal goat IgG ( F ). There was widespread moderate staining and scattered heavy staining of OSCC cells (cytoplasm and membrane), and light to moderate widespread staining of stromal cells ( G ). Staining was markedly reduced when the primary antibody was replaced with normal goat IgG ( H ). Each pair of positive and negative control images are at the same magnification. Scale bars are 50 μM long.
    Figure Legend Snippet: COX-2 and CD147 positive and negative controls for immunohistochemistry. ( A – D ) Photomicrographs of IHC staining using rabbit anti-COX-2 IgG (1:200) and normal rabbit IgG (1:200, negative control). The chromogen is DAB (brown), and the counterstain is hematoxylin (blue). The feline renal macula densa epithelial cells had an intense cytoplasmic signal ( A ) that was eliminated when the antibody was replaced with normal IgG ( B ). Scattered oral squamous cell carcinoma (OSCC) cells showed an intense cytoplasmic COX-2 signal ( C ), which was eliminated when the primary antibody was replaced with normal IgG ( D ). ( E , F ) IHC staining using goat anti-CD147 IgG (1:100) and normal goat IgG (1:100, negative control). Feline enterocytes showed moderately intense cytoplasmic and membranous signal ( E ), which was markedly reduced when the primary antibody was replaced with normal goat IgG ( F ). There was widespread moderate staining and scattered heavy staining of OSCC cells (cytoplasm and membrane), and light to moderate widespread staining of stromal cells ( G ). Staining was markedly reduced when the primary antibody was replaced with normal goat IgG ( H ). Each pair of positive and negative control images are at the same magnification. Scale bars are 50 μM long.

    Techniques Used: Immunohistochemistry, Staining, Negative Control

    COX-2 and CD147 expression varied between cases of feline OSCC. Photomicrographs of IHC staining using rabbit anti-COX-2 IgG (1:200) and goat anti-CD147 IgG (1:100). The chromogen is DAB (brown), and the counterstain is hematoxylin (blue). Case 1: There are scattered OSCC cells with an intense COX-2 signal ( A ). In contrast, OSCC cells showed widespread light to moderate CD147 signal ( B ). Case 2: This sample was COX-2 negative ( C ), but showed widespread moderate staining and scattered heavy staining for CD147 ( D ). Case 3: This sample was COX-2 negative ( E ) and CD147 negative in tumour cells ( F ), but had scattered moderate CD147 staining in the stroma ( F ). All images are at the same magnification. Scale bars are 100 μM long.
    Figure Legend Snippet: COX-2 and CD147 expression varied between cases of feline OSCC. Photomicrographs of IHC staining using rabbit anti-COX-2 IgG (1:200) and goat anti-CD147 IgG (1:100). The chromogen is DAB (brown), and the counterstain is hematoxylin (blue). Case 1: There are scattered OSCC cells with an intense COX-2 signal ( A ). In contrast, OSCC cells showed widespread light to moderate CD147 signal ( B ). Case 2: This sample was COX-2 negative ( C ), but showed widespread moderate staining and scattered heavy staining for CD147 ( D ). Case 3: This sample was COX-2 negative ( E ) and CD147 negative in tumour cells ( F ), but had scattered moderate CD147 staining in the stroma ( F ). All images are at the same magnification. Scale bars are 100 μM long.

    Techniques Used: Expressing, Immunohistochemistry, Staining

    Expression of COX-2 and CD147 in feline OSCC samples using an IHC grading system. ( A ) COX-2 staining results were obtained from 43 samples, 31 of which included adjacent oral epithelium. Each bar represents the percentage of cases that were considered positive for COX-2 expression within each compartment (tumor cells, stroma, and adjacent epithelium). Each bar is subdivided to demonstrate the percentage of cases assigned to each grade. COX-2 expression was highest in the tumor cells. ( B ) CD147 staining results were obtained from 43 samples, 28 of which included adjacent oral epithelium. CD147 expression was highest in the tumor cells. ( C ) Paired COX-2 and CD147 IHC results were available from 42 OSCC cases. Twenty-two (22) cases had positive COX-2 expression in the tumor cells (grades 1 and 2) and 20 cases were COX-2 negative in the tumor cells (grade 0). Each bar represents the percentage of positive CD147 cases within the COX-2 positive and COX-2 negative groups. There was no significant difference in CD147 expression between COX-2 positive and COX-2 negative cases. There were no cases with grade 5 staining. All of the statistical comparisons were made using Fisher’s exact test (* p value
    Figure Legend Snippet: Expression of COX-2 and CD147 in feline OSCC samples using an IHC grading system. ( A ) COX-2 staining results were obtained from 43 samples, 31 of which included adjacent oral epithelium. Each bar represents the percentage of cases that were considered positive for COX-2 expression within each compartment (tumor cells, stroma, and adjacent epithelium). Each bar is subdivided to demonstrate the percentage of cases assigned to each grade. COX-2 expression was highest in the tumor cells. ( B ) CD147 staining results were obtained from 43 samples, 28 of which included adjacent oral epithelium. CD147 expression was highest in the tumor cells. ( C ) Paired COX-2 and CD147 IHC results were available from 42 OSCC cases. Twenty-two (22) cases had positive COX-2 expression in the tumor cells (grades 1 and 2) and 20 cases were COX-2 negative in the tumor cells (grade 0). Each bar represents the percentage of positive CD147 cases within the COX-2 positive and COX-2 negative groups. There was no significant difference in CD147 expression between COX-2 positive and COX-2 negative cases. There were no cases with grade 5 staining. All of the statistical comparisons were made using Fisher’s exact test (* p value

    Techniques Used: Expressing, Immunohistochemistry, Staining

    23) Product Images from "Fucoidan protects hepatocytes from apoptosis and inhibits invasion of hepatocellular carcinoma by up-regulating p42/44 MAPK-dependent NDRG-1/CAP43"

    Article Title: Fucoidan protects hepatocytes from apoptosis and inhibits invasion of hepatocellular carcinoma by up-regulating p42/44 MAPK-dependent NDRG-1/CAP43

    Journal: Acta Pharmaceutica Sinica. B

    doi: 10.1016/j.apsb.2015.09.004

    Fucoidan inhibits liver metastasis in the distant metastasis model. (A) The number of intrahepatic metastases in the fucoidan treatment group and control group; (B) the sum of the maximal diameter of each nodule in the two groups; (C) histological analysis following fucoidan treatment. The nuclear grade of MH134 cells was determined by H E-staining and CD31 and GP-3 expression by IHC staining (200×).
    Figure Legend Snippet: Fucoidan inhibits liver metastasis in the distant metastasis model. (A) The number of intrahepatic metastases in the fucoidan treatment group and control group; (B) the sum of the maximal diameter of each nodule in the two groups; (C) histological analysis following fucoidan treatment. The nuclear grade of MH134 cells was determined by H E-staining and CD31 and GP-3 expression by IHC staining (200×).

    Techniques Used: Staining, Expressing, Immunohistochemistry

    24) Product Images from "Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model"

    Article Title: Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

    Journal: Canadian Journal of Veterinary Research

    doi:

    Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P
    Figure Legend Snippet: Results of enzyme-linked immunosorbent assay of the reaction between heat-inactivated isolates of K88ab + , K88ac + , K99, or Fas + ETEC and serum collected before and after vaccination of rabbits to produce antibodies specific to the recombinant fimbrial antigens. Data are expressed as mean optical density ± standard deviation (SD) from 3 experiments done in duplicate. Asterisks indicate a significant difference ( P

    Techniques Used: Enzyme-linked Immunosorbent Assay, Recombinant, Standard Deviation

    Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.
    Figure Legend Snippet: Results of Western blot analysis to identify recombinant K88ab, K88ac, K99, and FasA fimbrial antigens of enterotoxigenic Escherichia coli (ETEC) produced by JOL1285, JOL1286, JOL1287, and JOL1288, respectively, and expressed in the cell envelopes of Salmonella Typhimurium ghost bacteria. Lane c — control for each antigen.

    Techniques Used: Western Blot, Recombinant, Produced

    25) Product Images from "Affimer proteins are versatile and renewable affinity reagents"

    Article Title: Affimer proteins are versatile and renewable affinity reagents

    Journal: eLife

    doi: 10.7554/eLife.24903

    Use of tubulin binding Affimer in super-resolution microscopy. ( A ) Confocal images of microtubules in HeLa cells, stained with a rat α-tubulin antibody (YL1/2) which recognises tyrosinated tubulin, and an Affimer for polymerised tubulin, conjugated to Alexa Fluor 647. Images of an interphase and metaphase cell, together with an image of the cytokinetic furrow are shown. Arrows in the metaphase cell point to astral microtubules that are predominantly labelled with the antibody. Arrows in the cyokinetic furrow indicate the central region (Fleming body). Scale bar is 10 μm. ( B ) 3D dSTORM images of microtubules in a HeLa cell, labelled with Alexa Fluor 647 conjugated to a primary antibody to rat α-tubulin (left) and an Affimer for polymerised tubulin (right). These images are from separate cells. Localisations were aggregated into 10 nm bins and projected onto a single plane, with Gaussian smoothing. Scale bar 1 µm. ( C ) Intensity profile across the microtubule image labelled in ( B ) (yellow box), averaged along 510 nm of its length. The central decrease in intensity reflects the hollow structure of the microtubule. ( D ) Comparison of the average microtubule image intensity profile with antibody staining (dashed, mean of 6 microtubule sections), Affimer staining (solid, mean of 8 microtubule sections) and actual microtubule size (black circle). The FWHM of each average profile (as in ( C )) was found for a Gaussian fit and a Gaussian distribution is plotted here using the mean FWHM for each staining method. DOI: http://dx.doi.org/10.7554/eLife.24903.013
    Figure Legend Snippet: Use of tubulin binding Affimer in super-resolution microscopy. ( A ) Confocal images of microtubules in HeLa cells, stained with a rat α-tubulin antibody (YL1/2) which recognises tyrosinated tubulin, and an Affimer for polymerised tubulin, conjugated to Alexa Fluor 647. Images of an interphase and metaphase cell, together with an image of the cytokinetic furrow are shown. Arrows in the metaphase cell point to astral microtubules that are predominantly labelled with the antibody. Arrows in the cyokinetic furrow indicate the central region (Fleming body). Scale bar is 10 μm. ( B ) 3D dSTORM images of microtubules in a HeLa cell, labelled with Alexa Fluor 647 conjugated to a primary antibody to rat α-tubulin (left) and an Affimer for polymerised tubulin (right). These images are from separate cells. Localisations were aggregated into 10 nm bins and projected onto a single plane, with Gaussian smoothing. Scale bar 1 µm. ( C ) Intensity profile across the microtubule image labelled in ( B ) (yellow box), averaged along 510 nm of its length. The central decrease in intensity reflects the hollow structure of the microtubule. ( D ) Comparison of the average microtubule image intensity profile with antibody staining (dashed, mean of 6 microtubule sections), Affimer staining (solid, mean of 8 microtubule sections) and actual microtubule size (black circle). The FWHM of each average profile (as in ( C )) was found for a Gaussian fit and a Gaussian distribution is plotted here using the mean FWHM for each staining method. DOI: http://dx.doi.org/10.7554/eLife.24903.013

    Techniques Used: Binding Assay, Microscopy, Staining

    Affimer selection and specificity against TNT and DNT’s. ( A ) Chemical structures of TNBS, TNT, 2,3-DNT, 2,4-DNT and 2,6-DNT. ( B ) Phage ELISA results from 32 monoclonal Affimer reagents isolated against TNBS bound to ovalbumin. Binding specificity was also tested against TNBS bound to IgG and to unconjugated ovalbumin and IgG. ( C ) Competition ELISA of four TNT-Affimers to check specificity against a range of molecules across a concentration profile. Error bars = standard deviation from technical repeats of a representative ELISA. DOI: http://dx.doi.org/10.7554/eLife.24903.014
    Figure Legend Snippet: Affimer selection and specificity against TNT and DNT’s. ( A ) Chemical structures of TNBS, TNT, 2,3-DNT, 2,4-DNT and 2,6-DNT. ( B ) Phage ELISA results from 32 monoclonal Affimer reagents isolated against TNBS bound to ovalbumin. Binding specificity was also tested against TNBS bound to IgG and to unconjugated ovalbumin and IgG. ( C ) Competition ELISA of four TNT-Affimers to check specificity against a range of molecules across a concentration profile. Error bars = standard deviation from technical repeats of a representative ELISA. DOI: http://dx.doi.org/10.7554/eLife.24903.014

    Techniques Used: Selection, Enzyme-linked Immunosorbent Assay, Isolation, Binding Assay, Concentration Assay, Standard Deviation

    Use of HER4 binding Affimers in super-resolution imaging and single molecule tracking. ( A ) Phage ELISA for HER4 binding Affimers. ( B ) Average photon counts/pixel for HER4-binding Affimer labelled with CF640R and bound to CHO cells transfected with HER4 and to MCF7 cells expressing endogenous levels of HER4. ( C ) Wide field image of CHO cells transfected with HER4-CYT-eGFP showing localisation of HER4 via GFP fluorescence (top) and labelled with HER Affimer–Alexa647 (middle). The corresponding dSTORM image of HER4 Affimer conjugated to Alexa647 (bottom) with a 25 nm localisation precision. Scale bar = 2 μm. Right plots to show the number of molecules and cluster size of clusters identified by dSTORM. ( D ) Diffusion coefficients (left panel), and MSD curve (right panel) of HER4 Affimers labelled with CF640R and tracked on MCF7 cells expressing endogenous HER4. DOI: http://dx.doi.org/10.7554/eLife.24903.012
    Figure Legend Snippet: Use of HER4 binding Affimers in super-resolution imaging and single molecule tracking. ( A ) Phage ELISA for HER4 binding Affimers. ( B ) Average photon counts/pixel for HER4-binding Affimer labelled with CF640R and bound to CHO cells transfected with HER4 and to MCF7 cells expressing endogenous levels of HER4. ( C ) Wide field image of CHO cells transfected with HER4-CYT-eGFP showing localisation of HER4 via GFP fluorescence (top) and labelled with HER Affimer–Alexa647 (middle). The corresponding dSTORM image of HER4 Affimer conjugated to Alexa647 (bottom) with a 25 nm localisation precision. Scale bar = 2 μm. Right plots to show the number of molecules and cluster size of clusters identified by dSTORM. ( D ) Diffusion coefficients (left panel), and MSD curve (right panel) of HER4 Affimers labelled with CF640R and tracked on MCF7 cells expressing endogenous HER4. DOI: http://dx.doi.org/10.7554/eLife.24903.012

    Techniques Used: Binding Assay, Imaging, Enzyme-linked Immunosorbent Assay, Transfection, Expressing, Fluorescence, Diffusion-based Assay

    Characterisation of tenascin C (TNC) binding Affimer by affinity-histochemistry and ex vivo imaging of xenografts. ( A ) Phage ELISA for 48 monoclonal Affimers against TNC. The two controls are tenascin X (TNX) and streptavidin. ( B ) Immunohistochemistry of serial sections of a mouse xenograft (SW620 cell line), showing staining for TNC. Antibody/Affimer staining is shown as a light brown color with haemotoxylin counter staining (blue). ( C ) and ( D ) Mice were injected via their tail vein with rhodamine labelled TNC binding Affimer or a control GFP binding Affimer. After 24, 48, 72 and 96 hr the xenograft and organs were removed and visualized. ( C ) Organ images at 24 hr. ( D ) Quantification of rhodamine fluorescence (radiant efficiency in p/s/cm 2 /sr/μW/cm 2 ) ex vivo (n = 3). Mean background fluorescence intensity was normalized to sham injected control tumors and organs. DOI: http://dx.doi.org/10.7554/eLife.24903.009
    Figure Legend Snippet: Characterisation of tenascin C (TNC) binding Affimer by affinity-histochemistry and ex vivo imaging of xenografts. ( A ) Phage ELISA for 48 monoclonal Affimers against TNC. The two controls are tenascin X (TNX) and streptavidin. ( B ) Immunohistochemistry of serial sections of a mouse xenograft (SW620 cell line), showing staining for TNC. Antibody/Affimer staining is shown as a light brown color with haemotoxylin counter staining (blue). ( C ) and ( D ) Mice were injected via their tail vein with rhodamine labelled TNC binding Affimer or a control GFP binding Affimer. After 24, 48, 72 and 96 hr the xenograft and organs were removed and visualized. ( C ) Organ images at 24 hr. ( D ) Quantification of rhodamine fluorescence (radiant efficiency in p/s/cm 2 /sr/μW/cm 2 ) ex vivo (n = 3). Mean background fluorescence intensity was normalized to sham injected control tumors and organs. DOI: http://dx.doi.org/10.7554/eLife.24903.009

    Techniques Used: Binding Assay, Ex Vivo, Imaging, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Mouse Assay, Injection, Fluorescence

    Characterisation of TRPV1 binding Affimers. ( A ) Phage ELISA for 96 monoclonal Affimer reagents isolated against TRPV1 peptide. The negative control contained a different hydrophobic peptide sequence. ( B ) Affinity-cytochemistry on U2-OS cells transiently transfected with TRPV1 (TRPV1+) or control (TRPV1-) using Affimer 2. Binding was detected using an anti-HIS antibody fluorescently labeled with FITC. Binding of the Affimer is shown as a green and DAPI (a DNA stain) shown as blue (n = 3), ( C ) Co-localisation of Affimer staining with an anti-TRPV1 antibody. Antibody staining is shown in red. ( D ) A Flexstation was used to measure uptake of Fluo-4 AM, a calcium binding fluorescent small molecule, to measure calcium levels in capsaicin stimulated cells in the presence of Affimer control and TPRV1-binding Affimers (n = 3). DOI: http://dx.doi.org/10.7554/eLife.24903.008
    Figure Legend Snippet: Characterisation of TRPV1 binding Affimers. ( A ) Phage ELISA for 96 monoclonal Affimer reagents isolated against TRPV1 peptide. The negative control contained a different hydrophobic peptide sequence. ( B ) Affinity-cytochemistry on U2-OS cells transiently transfected with TRPV1 (TRPV1+) or control (TRPV1-) using Affimer 2. Binding was detected using an anti-HIS antibody fluorescently labeled with FITC. Binding of the Affimer is shown as a green and DAPI (a DNA stain) shown as blue (n = 3), ( C ) Co-localisation of Affimer staining with an anti-TRPV1 antibody. Antibody staining is shown in red. ( D ) A Flexstation was used to measure uptake of Fluo-4 AM, a calcium binding fluorescent small molecule, to measure calcium levels in capsaicin stimulated cells in the presence of Affimer control and TPRV1-binding Affimers (n = 3). DOI: http://dx.doi.org/10.7554/eLife.24903.008

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Isolation, Negative Control, Sequencing, Transfection, Labeling, Staining

    Ribbon diagrams of three crystal structures for Affimer (Adhiron) reagents. ( A ) X-ray crystal structure of Affimer scaffold (PDB ID no. 4N6T) at 1.75 A resolution. The amino acids from the loops connecting the four anti-parallel beta sheets are highlighted in pink. ( B ) Crystal structure of an Affimer against p300 (PDB ID no. 5A0O) ( C ) Crystal structure of an Affimer isolated against human SUMO proteins (PDB ID no. 5ELJ). The variable regions in B and C are shown in pink. DOI: http://dx.doi.org/10.7554/eLife.24903.003
    Figure Legend Snippet: Ribbon diagrams of three crystal structures for Affimer (Adhiron) reagents. ( A ) X-ray crystal structure of Affimer scaffold (PDB ID no. 4N6T) at 1.75 A resolution. The amino acids from the loops connecting the four anti-parallel beta sheets are highlighted in pink. ( B ) Crystal structure of an Affimer against p300 (PDB ID no. 5A0O) ( C ) Crystal structure of an Affimer isolated against human SUMO proteins (PDB ID no. 5ELJ). The variable regions in B and C are shown in pink. DOI: http://dx.doi.org/10.7554/eLife.24903.003

    Techniques Used: Isolation

    26) Product Images from "The PPAR? Agonist Efatutazone Increases the Spectrum of Well-Differentiated Mammary Cancer Subtypes Initiated by Loss of Full-Length BRCA1 in Association with TP53 Haploinsufficiency"

    Article Title: The PPAR? Agonist Efatutazone Increases the Spectrum of Well-Differentiated Mammary Cancer Subtypes Initiated by Loss of Full-Length BRCA1 in Association with TP53 Haploinsufficiency

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2013.02.006

    Levels of pAKT and expression of CDK6 are significantly reduced in cancers from efatutazone-treated Brca1 f11/f11/p53+/-/MMTV-Cre- mice. A : Stacked bar graphs summarizing IHC scores with representative IHC analysis images below for pAKT Ser473 and AKT
    Figure Legend Snippet: Levels of pAKT and expression of CDK6 are significantly reduced in cancers from efatutazone-treated Brca1 f11/f11/p53+/-/MMTV-Cre- mice. A : Stacked bar graphs summarizing IHC scores with representative IHC analysis images below for pAKT Ser473 and AKT

    Techniques Used: Expressing, Mouse Assay, Immunohistochemistry

    Expression patterns of PPARα, PPARγ, and RXRα in mammary tissue and cancers from control and efatutazone-treated mice. Representative images of H E-stained sections and PPARα, PPARγ, and RXRα IHC
    Figure Legend Snippet: Expression patterns of PPARα, PPARγ, and RXRα in mammary tissue and cancers from control and efatutazone-treated mice. Representative images of H E-stained sections and PPARα, PPARγ, and RXRα IHC

    Techniques Used: Expressing, Mouse Assay, Staining, Immunohistochemistry

    27) Product Images from "Patient-Derived Orthotopic Xenograft (PDOX) Mouse Models of Primary and Recurrent Meningioma"

    Article Title: Patient-Derived Orthotopic Xenograft (PDOX) Mouse Models of Primary and Recurrent Meningioma

    Journal: Cancers

    doi: 10.3390/cancers12061478

    Characteristics of PDOX models of primary and recurrent meningioma. ( A ) H E-stained paraffin sections of whole mouse brains bearing the intracranial xenograft tumors derived from the primary grade II (K29P) and recurrent grade III (K29R) meningiomas. ( B ) MRI showing the growth of cranial-base xenografts (red circles) of K29P-PDOX and obstructive hydrocephalus. ( C ). Kaplan–Meier survival curves for K29P and K29R PDOX models ( p = 0.0390, n = 8 out of 10 and 9 out of 9, respectively). ( D ). H E staining of xenograft and human tumors in the K29P and K29R-PDOX models. Areas of necrosis are circled in red. ( E ) . Representative images of IHC staining in matching patient tumor and PDOX models. MT: Mitochondria (human-specific). Ki-67: Cell proliferation. VWF: Von Willebrand factor (micro-vessels). VIM: Neurofilament vimentin. PDGFR1: EMA and platelet-derived growth factor receptor 1. Scale bars: ( A , B ) 500 mm, ( D , E ) 75 mm. Abbreviations: patient-derived orthotopic xenograft (PDOX); Hematoxylin and eosin (H E); magnetic resonance imaging (MRI); immunohistochemistry (IHC); epithelial membrane antigen (EMA).
    Figure Legend Snippet: Characteristics of PDOX models of primary and recurrent meningioma. ( A ) H E-stained paraffin sections of whole mouse brains bearing the intracranial xenograft tumors derived from the primary grade II (K29P) and recurrent grade III (K29R) meningiomas. ( B ) MRI showing the growth of cranial-base xenografts (red circles) of K29P-PDOX and obstructive hydrocephalus. ( C ). Kaplan–Meier survival curves for K29P and K29R PDOX models ( p = 0.0390, n = 8 out of 10 and 9 out of 9, respectively). ( D ). H E staining of xenograft and human tumors in the K29P and K29R-PDOX models. Areas of necrosis are circled in red. ( E ) . Representative images of IHC staining in matching patient tumor and PDOX models. MT: Mitochondria (human-specific). Ki-67: Cell proliferation. VWF: Von Willebrand factor (micro-vessels). VIM: Neurofilament vimentin. PDGFR1: EMA and platelet-derived growth factor receptor 1. Scale bars: ( A , B ) 500 mm, ( D , E ) 75 mm. Abbreviations: patient-derived orthotopic xenograft (PDOX); Hematoxylin and eosin (H E); magnetic resonance imaging (MRI); immunohistochemistry (IHC); epithelial membrane antigen (EMA).

    Techniques Used: Staining, Derivative Assay, Magnetic Resonance Imaging, Immunohistochemistry

    28) Product Images from "Proliferation, Adult Neuronal Stem Cells and Cells Migration in Pallium during Constitutive Neurogenesis and after Traumatic Injury of Telencephalon of Juvenile Masu Salmon, Oncorhynchus masou"

    Article Title: Proliferation, Adult Neuronal Stem Cells and Cells Migration in Pallium during Constitutive Neurogenesis and after Traumatic Injury of Telencephalon of Juvenile Masu Salmon, Oncorhynchus masou

    Journal: Brain Sciences

    doi: 10.3390/brainsci10040222

    Representative image of BrdU-immunolabeling in the pallial region of the telencephalon of the juvenile Oncorhynchus masou 1 week after the traumatic injury of the telencephalon. ( A ) On the left is a general view of the damaged hemisphere of the telencephalon; on the right, a fragment is partially shown of the intact hemisphere of the telencephalon; Dd—the dorsal zone; Dm—the medial; Dl—lateral; shown by red arrows, clusters of BrdU+ cells in PVZ (circled by a yellow dotted line), BrdU+ cells and nuclei in the area of injury are shown by black triangular arrows; ( B ) dorsal zone (Dd) at a higher magnification, the accumulation of BrdU+ cells in the PVZ is shown by yellow arrows (inset); ( C ) the medial zone (Dm) at a higher magnification, BrdU+ nuclei are shown by red arrows, yellow arrows show large BrdU+ cells in PVZ and PZ, the inset shows a PVZ fragment with a multilayer distribution of BrdU+ cells; ( D ) lateral zone (Dl) at a higher magnification, containing numerous clusters of BrdU+ cells in the PVZ, are shown by yellow arrows, in the inset an enlarged fragment of BrdU—immunopositive conglomerates (circled by yellow ovals), BrdU—cells are shown by black arrows; Immunohistochemical labeling of 5-brom-2-deoxy-uredine. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm. ( E ) The quantitative ratio of BrdU+ cells in intact animals (control group) and 1 week after traumatic injury ( n = 5 in each group, * p ≤ 0.05—significant difference from control groups. Student–Newman–Cales test; ( F ) the ratio of the optical density of BrdU-immunolabeling in the cells and nuclei of intact animals (control) and 1 week after traumatic injury (M ± SD) in Dd, Dl and Dm of the pallium of juvenile O. masou , UOP—the unit of optical density.
    Figure Legend Snippet: Representative image of BrdU-immunolabeling in the pallial region of the telencephalon of the juvenile Oncorhynchus masou 1 week after the traumatic injury of the telencephalon. ( A ) On the left is a general view of the damaged hemisphere of the telencephalon; on the right, a fragment is partially shown of the intact hemisphere of the telencephalon; Dd—the dorsal zone; Dm—the medial; Dl—lateral; shown by red arrows, clusters of BrdU+ cells in PVZ (circled by a yellow dotted line), BrdU+ cells and nuclei in the area of injury are shown by black triangular arrows; ( B ) dorsal zone (Dd) at a higher magnification, the accumulation of BrdU+ cells in the PVZ is shown by yellow arrows (inset); ( C ) the medial zone (Dm) at a higher magnification, BrdU+ nuclei are shown by red arrows, yellow arrows show large BrdU+ cells in PVZ and PZ, the inset shows a PVZ fragment with a multilayer distribution of BrdU+ cells; ( D ) lateral zone (Dl) at a higher magnification, containing numerous clusters of BrdU+ cells in the PVZ, are shown by yellow arrows, in the inset an enlarged fragment of BrdU—immunopositive conglomerates (circled by yellow ovals), BrdU—cells are shown by black arrows; Immunohistochemical labeling of 5-brom-2-deoxy-uredine. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm. ( E ) The quantitative ratio of BrdU+ cells in intact animals (control group) and 1 week after traumatic injury ( n = 5 in each group, * p ≤ 0.05—significant difference from control groups. Student–Newman–Cales test; ( F ) the ratio of the optical density of BrdU-immunolabeling in the cells and nuclei of intact animals (control) and 1 week after traumatic injury (M ± SD) in Dd, Dl and Dm of the pallium of juvenile O. masou , UOP—the unit of optical density.

    Techniques Used: Immunolabeling, Immunohistochemistry, Labeling

    Representative image of Vim-immunolabeling in the pallial region of the intact telencephalon ( A ) of juvenile Oncorhynchus masou and 1 week after traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of Vim in intact pallium, immunopositive cells (inset, red arrows) and granules (blue arrow) in the parenchymal zone (PZ), Vim- cells (black arrow); ( B ) Vim+ cell complexes (inset) and clusters (red dashed oval) of intensely Vim-labeled cells (red arrow) and moderately Vim-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, radial glia fiber (white arrows), Vim+ granules (blue arrow); ( C ) post-traumatic reorganization of Vim-immunopositivity in Dl, Vim+ heterogeneous complexes in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with immuno-negative regions, radial glia fibers are limited by a red dashed rectangle, intensely Vim-labeled cells are shown by red arrows, moderately Vim- labeled cells (yellow arrow); ( D ) weakly labeled Vim+ cells after injury in the PVZ of Dm (inset), moderately labeled cells (yellow arrow), intensely labeled granules (red arrows), Vim- cell (black arrow) and radial glia (white arrow); ( E ) the quantitative ratio of Vim+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of vimentin in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.
    Figure Legend Snippet: Representative image of Vim-immunolabeling in the pallial region of the intact telencephalon ( A ) of juvenile Oncorhynchus masou and 1 week after traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of Vim in intact pallium, immunopositive cells (inset, red arrows) and granules (blue arrow) in the parenchymal zone (PZ), Vim- cells (black arrow); ( B ) Vim+ cell complexes (inset) and clusters (red dashed oval) of intensely Vim-labeled cells (red arrow) and moderately Vim-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, radial glia fiber (white arrows), Vim+ granules (blue arrow); ( C ) post-traumatic reorganization of Vim-immunopositivity in Dl, Vim+ heterogeneous complexes in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with immuno-negative regions, radial glia fibers are limited by a red dashed rectangle, intensely Vim-labeled cells are shown by red arrows, moderately Vim- labeled cells (yellow arrow); ( D ) weakly labeled Vim+ cells after injury in the PVZ of Dm (inset), moderately labeled cells (yellow arrow), intensely labeled granules (red arrows), Vim- cell (black arrow) and radial glia (white arrow); ( E ) the quantitative ratio of Vim+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of vimentin in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.

    Techniques Used: Immunolabeling, Labeling, Immunohistochemistry, Staining

    Representative image of BrdU-immunolabeling in the pallial region of the telencephalon of an intact juvenile Oncorhynchus masou . ( A ) General view of the telencephalon, the pictogram shows the zones of the dorsal telencephalon (pallium), Dd-dorsal, Dm-medial, Dl-lateral, the inset (outlined by a red rectangle) shows a fragment including PVZ-periventricular zone and SVZ-subventricular zone, BrdU+ cells are shown by red arrows, BrdU-cells are indicated by black arrows; ( B ) dorsal zone (Dd) at a higher magnification, accumulation of BrdU+ nuclei in the red oval, PZ-parenchymal zone of the telencephalon, for other designations, see Figure 1 A; ( C )-lateral zone (Dl) at a higher magnification, migrating tangentially elongated BrdU+ cells are shown by red arrows, BrdU+ cells in the surface layer of the PVZ are shown in the inset (in the red box); ( D ) medial zone (Dm), the group of radially migrating BrdU+ cells is shown in the inset (in the red rectangle), the cluster of BrdU+ nuclei in the yellow oval. Immunohistochemical labeling of 5-brom-2-deoxy-uredine. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm. ( E ) The comparative distribution of BrdU+ cells in various regions of intact pallium. ( n = 5 in each group; #-significant intergroup differences). One-way analysis of variance (ANOVA); ( F ) the optical density of BrdU immunolabeling in cells of various types and nuclei (M ± SD) of Dd, Dl, and Dm of pallium of juvenile O. masou , UOP—the units of optical density.
    Figure Legend Snippet: Representative image of BrdU-immunolabeling in the pallial region of the telencephalon of an intact juvenile Oncorhynchus masou . ( A ) General view of the telencephalon, the pictogram shows the zones of the dorsal telencephalon (pallium), Dd-dorsal, Dm-medial, Dl-lateral, the inset (outlined by a red rectangle) shows a fragment including PVZ-periventricular zone and SVZ-subventricular zone, BrdU+ cells are shown by red arrows, BrdU-cells are indicated by black arrows; ( B ) dorsal zone (Dd) at a higher magnification, accumulation of BrdU+ nuclei in the red oval, PZ-parenchymal zone of the telencephalon, for other designations, see Figure 1 A; ( C )-lateral zone (Dl) at a higher magnification, migrating tangentially elongated BrdU+ cells are shown by red arrows, BrdU+ cells in the surface layer of the PVZ are shown in the inset (in the red box); ( D ) medial zone (Dm), the group of radially migrating BrdU+ cells is shown in the inset (in the red rectangle), the cluster of BrdU+ nuclei in the yellow oval. Immunohistochemical labeling of 5-brom-2-deoxy-uredine. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm. ( E ) The comparative distribution of BrdU+ cells in various regions of intact pallium. ( n = 5 in each group; #-significant intergroup differences). One-way analysis of variance (ANOVA); ( F ) the optical density of BrdU immunolabeling in cells of various types and nuclei (M ± SD) of Dd, Dl, and Dm of pallium of juvenile O. masou , UOP—the units of optical density.

    Techniques Used: Immunolabeling, Immunohistochemistry, Labeling

    Representative image of doublecortin DC-immunolabeling in the pallial region of the intact telencephalon ( A ) of the juvenile Oncorhynchus masou and 1 week after the traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of DC in intact pallium, immunopositive cells (inset, red arrows), radial glia (black arrows) and granules (white arrows) in the parenchymal zone (PZ); ( B ) DC+ cells (inset) and clusters (white dashed oval) of intensely DC-labeled cells (red arrow) and moderately DC-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, Vim+ granules (blue arrow); ( C ) post-traumatic expression of DC immunopositivity in Dl, DC+ heterogeneous complexes in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with moderately-labeled areas, accumulation of intensely DC-labeled cells (in a yellow dashed oval) other designations as in Figure 5 B; ( D ) patterns of DC-immunopositivity after trauma in PVZ and SVZ Dm (inset), intensely labeled cells (red arrows), moderately labeled cells (yellow arrows), clusters of intensely labeled cells (in a yellow oval); ( E ) the quantitative ratio of DC+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; ** p ≤ 0.01; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of doublecortin in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.
    Figure Legend Snippet: Representative image of doublecortin DC-immunolabeling in the pallial region of the intact telencephalon ( A ) of the juvenile Oncorhynchus masou and 1 week after the traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of DC in intact pallium, immunopositive cells (inset, red arrows), radial glia (black arrows) and granules (white arrows) in the parenchymal zone (PZ); ( B ) DC+ cells (inset) and clusters (white dashed oval) of intensely DC-labeled cells (red arrow) and moderately DC-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, Vim+ granules (blue arrow); ( C ) post-traumatic expression of DC immunopositivity in Dl, DC+ heterogeneous complexes in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with moderately-labeled areas, accumulation of intensely DC-labeled cells (in a yellow dashed oval) other designations as in Figure 5 B; ( D ) patterns of DC-immunopositivity after trauma in PVZ and SVZ Dm (inset), intensely labeled cells (red arrows), moderately labeled cells (yellow arrows), clusters of intensely labeled cells (in a yellow oval); ( E ) the quantitative ratio of DC+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; ** p ≤ 0.01; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of doublecortin in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.

    Techniques Used: Immunolabeling, Labeling, Expressing, Immunohistochemistry, Staining

    Representative image of GFAP-immunolabeling in the pallial region of the intact telencephalon ( A ) of juvenile Oncorhynchus masou and 1 week after traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of GFAP in intact pallium, immunopositive cells (red arrows) and their clusters (blue dashed line) in PVZ (inset in the black rectangle) and radial glia fibers (white arrows) in the SVZ (inset in the red rectangle); ( B ) GFAP+ cells complexes (inset in red rectangle) and clusters (inset in black rectangle) of intensely GFAP-labeled cells (red star) and moderately GFAP-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, radial glia fibers bounded by white lines, GFAP+ granules (blue arrow); ( C ) post-traumatic reorganization of GFAP immunopositivity in Dl, GFAP+ heterogeneous complexes (in a white dashed oval) in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with immuno-negative regions, radial glia fibers are bounded by a white segment, GFAP+ cells are shown by red arrows; ( D ) heterogeneous GFAP+ cells after trauma in the PVZ of Dm (inset), moderately labeled (yellow arrows), intensely labeled (red arrows) and radial glia (white arrows); ( E ) quantitative ratio of GFAP+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; ** p ≤ 0.01; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of glial fibrillar acidic protein in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.
    Figure Legend Snippet: Representative image of GFAP-immunolabeling in the pallial region of the intact telencephalon ( A ) of juvenile Oncorhynchus masou and 1 week after traumatic injury ( B – D ) of the telencephalon. ( A ) Localization of GFAP in intact pallium, immunopositive cells (red arrows) and their clusters (blue dashed line) in PVZ (inset in the black rectangle) and radial glia fibers (white arrows) in the SVZ (inset in the red rectangle); ( B ) GFAP+ cells complexes (inset in red rectangle) and clusters (inset in black rectangle) of intensely GFAP-labeled cells (red star) and moderately GFAP-labeled cell (yellow arrow) in PVZ of Dd, 1 week after damage to the telencephalon, radial glia fibers bounded by white lines, GFAP+ granules (blue arrow); ( C ) post-traumatic reorganization of GFAP immunopositivity in Dl, GFAP+ heterogeneous complexes (in a white dashed oval) in PVZ and SVZ (an enlarged fragment is shown in the inset), alternating with immuno-negative regions, radial glia fibers are bounded by a white segment, GFAP+ cells are shown by red arrows; ( D ) heterogeneous GFAP+ cells after trauma in the PVZ of Dm (inset), moderately labeled (yellow arrows), intensely labeled (red arrows) and radial glia (white arrows); ( E ) quantitative ratio of GFAP+ cells in intact animals (control group) and 1 week after traumatic injury to the telencephalon ( n = 5 in each group, * p ≤ 0.05; ** p ≤ 0.01; significant difference from control groups). Student–Newman–Cales test. Immunohistochemical labeling of glial fibrillar acidic protein in combination with methyl green staining. Scale bar: in ( A ) 200 μm, ( B – D ) 100 μm.

    Techniques Used: Immunolabeling, Labeling, Immunohistochemistry, Staining

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    Immunohistochemistry:

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    Incubation:

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    Plasmid Preparation:

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    Vector Laboratories immunohistochemical analysis immunostaining
    Effects of follistatin on expression of α -SMA in kidneys after UUO. (a) Expression of α -SMA, a marker for myofibroblasts, in the UUO kidneys, was examined by <t>immunostaining.</t> (A) Normal kidney. (B) Contralateral kidneys, 10 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 10 days. (F) Follistatin-treated UUO kidneys, 10 days. Magnification: ×200. α -SMA (green), DAPI (blue). (b) Quantitative analysis of α -SMA-positive area. α -SMA-positive area in the kidneys at 10 days after UUO was assessed as described in Section 2 . Values are mean ± SE ( n = 5). * P
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    Effects of follistatin on expression of α -SMA in kidneys after UUO. (a) Expression of α -SMA, a marker for myofibroblasts, in the UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 10 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 10 days. (F) Follistatin-treated UUO kidneys, 10 days. Magnification: ×200. α -SMA (green), DAPI (blue). (b) Quantitative analysis of α -SMA-positive area. α -SMA-positive area in the kidneys at 10 days after UUO was assessed as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Journal: BioMed Research International

    Article Title: Follistatin, an Activin Antagonist, Ameliorates Renal Interstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction

    doi: 10.1155/2014/376191

    Figure Lengend Snippet: Effects of follistatin on expression of α -SMA in kidneys after UUO. (a) Expression of α -SMA, a marker for myofibroblasts, in the UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 10 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 10 days. (F) Follistatin-treated UUO kidneys, 10 days. Magnification: ×200. α -SMA (green), DAPI (blue). (b) Quantitative analysis of α -SMA-positive area. α -SMA-positive area in the kidneys at 10 days after UUO was assessed as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Article Snippet: Immunohistochemical Analysis Immunostaining with the avidin-biotin coupling immunoperoxidase technique was performed using a Vectastain Elite ABC kit (Vector Laboratories, Burlingame, CA) in accordance with the manufacturer's instructions.

    Techniques: Expressing, Marker, Immunostaining

    Effects of follistatin on the production of extracellular matrix in kidneys after UUO. (a) Production of type I collagen (A–D), type III collagen (E–H), and fibronectin (I–L) in the UUO kidneys was examined by immunostaining. (A, E, I) normal kidney. (B, F, J) contralateral kidney. (C, G, K) saline-treated UUO kidney, 7 days. (D, H, L) follistatin-treated UUO kidney, 7 days. Type I collagen, type III collagen, and fibronectin (green). Magnification: ×100. (b) Quantitative analysis of extracellular matrix production. Type I collagen, type III collagen, and fibronectin-positive area in kidneys at 7 days after UUO was measured as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Journal: BioMed Research International

    Article Title: Follistatin, an Activin Antagonist, Ameliorates Renal Interstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction

    doi: 10.1155/2014/376191

    Figure Lengend Snippet: Effects of follistatin on the production of extracellular matrix in kidneys after UUO. (a) Production of type I collagen (A–D), type III collagen (E–H), and fibronectin (I–L) in the UUO kidneys was examined by immunostaining. (A, E, I) normal kidney. (B, F, J) contralateral kidney. (C, G, K) saline-treated UUO kidney, 7 days. (D, H, L) follistatin-treated UUO kidney, 7 days. Type I collagen, type III collagen, and fibronectin (green). Magnification: ×100. (b) Quantitative analysis of extracellular matrix production. Type I collagen, type III collagen, and fibronectin-positive area in kidneys at 7 days after UUO was measured as described in Section 2 . Values are mean ± SE ( n = 5). * P

    Article Snippet: Immunohistochemical Analysis Immunostaining with the avidin-biotin coupling immunoperoxidase technique was performed using a Vectastain Elite ABC kit (Vector Laboratories, Burlingame, CA) in accordance with the manufacturer's instructions.

    Techniques: Immunostaining

    Effects of follistatin on macrophage infiltration in kidneys after UUO. (a) Expression of CD68, a marker for macrophages, in UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 3 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 7 days. (F) Follistatin-treated UUO kidneys, 7 days. CD68-positive cells (brown). Magnification: ×400. (b) Quantitative analysis of CD68-positive cell number. CD68-positive cells were counted in 10 randomly selected fields per rat at ×400 magnification. Values are mean ± SE ( n = 5). Saline (white bars), follistatin (black bars). * P

    Journal: BioMed Research International

    Article Title: Follistatin, an Activin Antagonist, Ameliorates Renal Interstitial Fibrosis in a Rat Model of Unilateral Ureteral Obstruction

    doi: 10.1155/2014/376191

    Figure Lengend Snippet: Effects of follistatin on macrophage infiltration in kidneys after UUO. (a) Expression of CD68, a marker for macrophages, in UUO kidneys, was examined by immunostaining. (A) Normal kidney. (B) Contralateral kidneys, 3 days. (C) Saline-treated UUO kidneys, 3 days. (D) Follistatin-treated UUO kidneys, 3 days. (E) Saline-treated UUO kidneys, 7 days. (F) Follistatin-treated UUO kidneys, 7 days. CD68-positive cells (brown). Magnification: ×400. (b) Quantitative analysis of CD68-positive cell number. CD68-positive cells were counted in 10 randomly selected fields per rat at ×400 magnification. Values are mean ± SE ( n = 5). Saline (white bars), follistatin (black bars). * P

    Article Snippet: Immunohistochemical Analysis Immunostaining with the avidin-biotin coupling immunoperoxidase technique was performed using a Vectastain Elite ABC kit (Vector Laboratories, Burlingame, CA) in accordance with the manufacturer's instructions.

    Techniques: Expressing, Marker, Immunostaining

    Study 2 (monthly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (week 23 postvaccination). The higher magnification

    Journal:

    Article Title: Therapeutic Vaccination against Helicobacter pylori in the Beagle Dog Experimental Model: Safety, Immunogenicity, and Efficacy

    doi: 10.1128/IAI.72.6.3252-3259.2004

    Figure Lengend Snippet: Study 2 (monthly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (week 23 postvaccination). The higher magnification

    Article Snippet: Adjacent sections were subjected to immunohistochemical (IHC) analysis with a mouse anti-VacA monoclonal antibody as previously described ( ); the antibody binding was revealed by using biotinylated anti-mouse secondary antibody and avidin-biotin complex-peroxidase (Vector Laboratories, Burlingame, Calif.) with 3-3′diaminobenzidine-hydrochloride (Sigma Chemical Co., St. Louis, Mo.) as a chromogen substrate.

    Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Sampling

    Study 1 (weekly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (29th week postvaccination). The higher magnification

    Journal:

    Article Title: Therapeutic Vaccination against Helicobacter pylori in the Beagle Dog Experimental Model: Safety, Immunogenicity, and Efficacy

    doi: 10.1128/IAI.72.6.3252-3259.2004

    Figure Lengend Snippet: Study 1 (weekly immunization): representative IHC results obtained with anti-VacA antibody on 4-μm-thick sections of formalin-fixed, paraffin-embedded gastric antral biopsies at the last sampling (29th week postvaccination). The higher magnification

    Article Snippet: Adjacent sections were subjected to immunohistochemical (IHC) analysis with a mouse anti-VacA monoclonal antibody as previously described ( ); the antibody binding was revealed by using biotinylated anti-mouse secondary antibody and avidin-biotin complex-peroxidase (Vector Laboratories, Burlingame, Calif.) with 3-3′diaminobenzidine-hydrochloride (Sigma Chemical Co., St. Louis, Mo.) as a chromogen substrate.

    Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Sampling

    Butyrate inhibition of colonic epithelial cell growth is dependent on induction of Hsp25. ( a ) Butyrate inhibition of cell growth (clear bar, fourth from left) is blocked by silencing Hsp25 (grey bar, far right), but not by scrambled (non-sense) siRNA (siScr). Cell proliferation was measured by the WST-1 assay. (b) Lentiviral-induced expression of Hsp25 (Hsp25 lenti) in mouse colon decreases expression of the proliferation marker Ki-67 (brown staining). As a control, empty cassette GFP lentivector (GFP-lenti) was administered the same way. After one hour exposure of the colonic mucosa to luminally-administered lentivirus, the colonic mucosal segment was marked, returned to the abdominal cavity, and then harvested 3 days later, as described in methods. (c) Upper panel : Alpha smooth muscle actin staining was used to identify peri-crypt myofibroblasts (brown immunostaining). Lower panel: Distribution of pericrypt myofibroblast, identified by alpha smooth muscle actin immunostaining, among the upper, middle, and lower tritiles (thirds) of colonic crypts. (d) Intestinal myofibroblast-derived conditioned medium (CM) blocks butyrate-stimulated (1 and 5 mM) Hsp25 expression in intestinal epithelial YAMC monolayers. No effects were seen on Hsc70 protein expression which is used as a loading control (d, upper panel). *p

    Journal: Scientific Reports

    Article Title: Butyrate and bioactive proteolytic form of Wnt-5a regulate colonic epithelial proliferation and spatial development

    doi: 10.1038/srep32094

    Figure Lengend Snippet: Butyrate inhibition of colonic epithelial cell growth is dependent on induction of Hsp25. ( a ) Butyrate inhibition of cell growth (clear bar, fourth from left) is blocked by silencing Hsp25 (grey bar, far right), but not by scrambled (non-sense) siRNA (siScr). Cell proliferation was measured by the WST-1 assay. (b) Lentiviral-induced expression of Hsp25 (Hsp25 lenti) in mouse colon decreases expression of the proliferation marker Ki-67 (brown staining). As a control, empty cassette GFP lentivector (GFP-lenti) was administered the same way. After one hour exposure of the colonic mucosa to luminally-administered lentivirus, the colonic mucosal segment was marked, returned to the abdominal cavity, and then harvested 3 days later, as described in methods. (c) Upper panel : Alpha smooth muscle actin staining was used to identify peri-crypt myofibroblasts (brown immunostaining). Lower panel: Distribution of pericrypt myofibroblast, identified by alpha smooth muscle actin immunostaining, among the upper, middle, and lower tritiles (thirds) of colonic crypts. (d) Intestinal myofibroblast-derived conditioned medium (CM) blocks butyrate-stimulated (1 and 5 mM) Hsp25 expression in intestinal epithelial YAMC monolayers. No effects were seen on Hsc70 protein expression which is used as a loading control (d, upper panel). *p

    Article Snippet: Immunohistochemical staining Immunohistochemical staining was performed on sections from formalin-fixed, paraffin-embedded mouse colon for smooth muscle actin, Hsp25, and Wnt-5a or Ki-67 using the Vectastain Elite ABC kit (Vector Labs, Burlingame, CA) or EnVision+System (DakoCytomation, Australia).

    Techniques: Inhibition, WST-1 Assay, Expressing, Marker, Staining, Immunostaining, Derivative Assay

    Localization of A subunit for Activin, NGAL, and KIM-1 in the Kidneys after Renal Ischemia. ( A ) Localization of βA subunit for activin and NGAL in the kidneys after renal ischemia for 25 min was examined by immunostaining. βA subunit (green), NGAL (red). Magnification: ×400. ( B ) Localization of βA subunit for activin and KIM-1 in the kidneys after renal ischemia was examined by immunostaining. βA subunit (green), KIM-1 (red). Magnification: ×1000. ( C ) Localization of βA subunit for activin and PCNA in the ischemic kidneys at 48 hr after reperfusion. βA subunit (red), PCNA (green), and DAPI (blue). Magnification: ×400. ( D ) Immunostaining of βA subunit for activin (a, b) and TUNEL staining (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’).Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000. ( E ) Immunostaining of βA subunit for activin (a,b) and caspase 3 (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’). Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000.

    Journal: Scientific Reports

    Article Title: Identification of Urinary Activin A as a Novel Biomarker Reflecting the Severity of Acute Kidney Injury

    doi: 10.1038/s41598-018-23564-3

    Figure Lengend Snippet: Localization of A subunit for Activin, NGAL, and KIM-1 in the Kidneys after Renal Ischemia. ( A ) Localization of βA subunit for activin and NGAL in the kidneys after renal ischemia for 25 min was examined by immunostaining. βA subunit (green), NGAL (red). Magnification: ×400. ( B ) Localization of βA subunit for activin and KIM-1 in the kidneys after renal ischemia was examined by immunostaining. βA subunit (green), KIM-1 (red). Magnification: ×1000. ( C ) Localization of βA subunit for activin and PCNA in the ischemic kidneys at 48 hr after reperfusion. βA subunit (red), PCNA (green), and DAPI (blue). Magnification: ×400. ( D ) Immunostaining of βA subunit for activin (a, b) and TUNEL staining (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’).Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000. ( E ) Immunostaining of βA subunit for activin (a,b) and caspase 3 (a’,b’) in the ischemic kidneys at 48 hr after reperfusion using serial sections (a-a’,b-b’). Positive signals (brown). PAS-positive brush border (red). Magnification: ×1000.

    Article Snippet: Immunohistochemical Analysis Immunostaining was performed using a VECTASTAIN ABC-kit (Vector Laboratories) as described previously .

    Techniques: Immunostaining, TUNEL Assay, Staining