endogenous manf 1 transcripts  (Qiagen)

 
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    Name:
    RT² qPCR Primer Assay
    Description:
    RT² qPCR Primer Assays are specifically designed and experimentally verified for real time PCR analysis The rigorous assay verification criteria ensure PCR specificity and efficiency for reliable and accurate gene expression analysis results
    Catalog Number:
    330001
    Price:
    144
    Category:
    Assay PCR qPCR
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    Structured Review

    Qiagen endogenous manf 1 transcripts
    RT² qPCR Primer Assay
    RT² qPCR Primer Assays are specifically designed and experimentally verified for real time PCR analysis The rigorous assay verification criteria ensure PCR specificity and efficiency for reliable and accurate gene expression analysis results
    https://www.bioz.com/result/endogenous manf 1 transcripts/product/Qiagen
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    endogenous manf 1 transcripts - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "C. elegans MANF Homolog Is Necessary for the Protection of Dopaminergic Neurons and ER Unfolded Protein Response"

    Article Title: C. elegans MANF Homolog Is Necessary for the Protection of Dopaminergic Neurons and ER Unfolded Protein Response

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00544

    Expression profile of manf-1 and fecundity defects in manf-1 mutants. (A) qRT-PCR data showing a decline in endogenous manf-1 transcript levels with age in wildtype animals. Results are means of experiments performed in duplicate ± SEMs, normalized to day-1 adult manf-1 expression levels, ** p
    Figure Legend Snippet: Expression profile of manf-1 and fecundity defects in manf-1 mutants. (A) qRT-PCR data showing a decline in endogenous manf-1 transcript levels with age in wildtype animals. Results are means of experiments performed in duplicate ± SEMs, normalized to day-1 adult manf-1 expression levels, ** p

    Techniques Used: Expressing, Quantitative RT-PCR

    Characterization of manf-1 alleles. (A) manf-1 cDNA from wildtype (N2) and mutant ( tm3603 ). 500 bp DNA ladder is marked for size comparison. (B) qRT-PCR analysis of manf-1 in 1 day old adults. Results are means of experiments performed in triplicate ± SEMs, ** p
    Figure Legend Snippet: Characterization of manf-1 alleles. (A) manf-1 cDNA from wildtype (N2) and mutant ( tm3603 ). 500 bp DNA ladder is marked for size comparison. (B) qRT-PCR analysis of manf-1 in 1 day old adults. Results are means of experiments performed in triplicate ± SEMs, ** p

    Techniques Used: Mutagenesis, Quantitative RT-PCR

    Dopaminergic neuron and ER stress phenotypes of manf-1 mutants. Quantification of dopaminergic neuronal defects in day-1 to day-9 old adults of tm3603 (A) and gk3677 (B) mutant animals. In each case wildtype (WT) animals were used as controls. Experiments performed in duplicate ± SEMs, * p
    Figure Legend Snippet: Dopaminergic neuron and ER stress phenotypes of manf-1 mutants. Quantification of dopaminergic neuronal defects in day-1 to day-9 old adults of tm3603 (A) and gk3677 (B) mutant animals. In each case wildtype (WT) animals were used as controls. Experiments performed in duplicate ± SEMs, * p

    Techniques Used: Mutagenesis

    manf-1 mutants show enhanced aggregation of α-Synuclein. (A) Expression analysis of human SNCA::YFP gene within the body wall muscles of wildtype (N2) and tm3603 animals. (B) Maximum projection of Z-stack images at days 1 and 5 of adulthood showing YFP fluorescent puncta. (C) Number of puncta of diameter 2 μm and above determined by manual counting and automated analysis of total pixel area after thresholding. Experiments performed in triplicate ± SEMs, * p
    Figure Legend Snippet: manf-1 mutants show enhanced aggregation of α-Synuclein. (A) Expression analysis of human SNCA::YFP gene within the body wall muscles of wildtype (N2) and tm3603 animals. (B) Maximum projection of Z-stack images at days 1 and 5 of adulthood showing YFP fluorescent puncta. (C) Number of puncta of diameter 2 μm and above determined by manual counting and automated analysis of total pixel area after thresholding. Experiments performed in triplicate ± SEMs, * p

    Techniques Used: Expressing

    MANF and CDNF homologs in selected organisms. (A) Protein domains and structure of the C. elegans manf-1 protein. (B) Schematic representation of MANF and CDNF proteins from Caenorhabditis elegans (Ce), Drosophila melanogaster ( Dm ), Homo sapiens ( Hs ) and Mus musculus ( Mm ) with percent identity and similarity indicated relative to C. elegans manf-1 .
    Figure Legend Snippet: MANF and CDNF homologs in selected organisms. (A) Protein domains and structure of the C. elegans manf-1 protein. (B) Schematic representation of MANF and CDNF proteins from Caenorhabditis elegans (Ce), Drosophila melanogaster ( Dm ), Homo sapiens ( Hs ) and Mus musculus ( Mm ) with percent identity and similarity indicated relative to C. elegans manf-1 .

    Techniques Used:

    2) Product Images from "Dual fluorescence detection of protein and RNA in Drosophila tissues"

    Article Title: Dual fluorescence detection of protein and RNA in Drosophila tissues

    Journal: Nature protocols

    doi: 10.1038/nprot.2012.105

    Dual labeling of let-7 miRNA and Fas3 protein in the testis. An LNA- let-7- DIG-labeled probe was used to detect let-7 miRNA expression in adult testes from 1-d-old male flies (genotype: updGAL4 ; UAS- let7 701.12.9 ). ( a – c ) Fas3 ( a ), let-7 ( b ) and
    Figure Legend Snippet: Dual labeling of let-7 miRNA and Fas3 protein in the testis. An LNA- let-7- DIG-labeled probe was used to detect let-7 miRNA expression in adult testes from 1-d-old male flies (genotype: updGAL4 ; UAS- let7 701.12.9 ). ( a – c ) Fas3 ( a ), let-7 ( b ) and

    Techniques Used: Labeling, Expressing

    3) Product Images from "C. elegans MANF Homolog Is Necessary for the Protection of Dopaminergic Neurons and ER Unfolded Protein Response"

    Article Title: C. elegans MANF Homolog Is Necessary for the Protection of Dopaminergic Neurons and ER Unfolded Protein Response

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00544

    Expression profile of manf-1 and fecundity defects in manf-1 mutants. (A) qRT-PCR data showing a decline in endogenous manf-1 transcript levels with age in wildtype animals. Results are means of experiments performed in duplicate ± SEMs, normalized to day-1 adult manf-1 expression levels, ** p
    Figure Legend Snippet: Expression profile of manf-1 and fecundity defects in manf-1 mutants. (A) qRT-PCR data showing a decline in endogenous manf-1 transcript levels with age in wildtype animals. Results are means of experiments performed in duplicate ± SEMs, normalized to day-1 adult manf-1 expression levels, ** p

    Techniques Used: Expressing, Quantitative RT-PCR

    Characterization of manf-1 alleles. (A) manf-1 cDNA from wildtype (N2) and mutant ( tm3603 ). 500 bp DNA ladder is marked for size comparison. (B) qRT-PCR analysis of manf-1 in 1 day old adults. Results are means of experiments performed in triplicate ± SEMs, ** p
    Figure Legend Snippet: Characterization of manf-1 alleles. (A) manf-1 cDNA from wildtype (N2) and mutant ( tm3603 ). 500 bp DNA ladder is marked for size comparison. (B) qRT-PCR analysis of manf-1 in 1 day old adults. Results are means of experiments performed in triplicate ± SEMs, ** p

    Techniques Used: Mutagenesis, Quantitative RT-PCR

    Dopaminergic neuron and ER stress phenotypes of manf-1 mutants. Quantification of dopaminergic neuronal defects in day-1 to day-9 old adults of tm3603 (A) and gk3677 (B) mutant animals. In each case wildtype (WT) animals were used as controls. Experiments performed in duplicate ± SEMs, * p
    Figure Legend Snippet: Dopaminergic neuron and ER stress phenotypes of manf-1 mutants. Quantification of dopaminergic neuronal defects in day-1 to day-9 old adults of tm3603 (A) and gk3677 (B) mutant animals. In each case wildtype (WT) animals were used as controls. Experiments performed in duplicate ± SEMs, * p

    Techniques Used: Mutagenesis

    manf-1 mutants show enhanced aggregation of α-Synuclein. (A) Expression analysis of human SNCA::YFP gene within the body wall muscles of wildtype (N2) and tm3603 animals. (B) Maximum projection of Z-stack images at days 1 and 5 of adulthood showing YFP fluorescent puncta. (C) Number of puncta of diameter 2 μm and above determined by manual counting and automated analysis of total pixel area after thresholding. Experiments performed in triplicate ± SEMs, * p
    Figure Legend Snippet: manf-1 mutants show enhanced aggregation of α-Synuclein. (A) Expression analysis of human SNCA::YFP gene within the body wall muscles of wildtype (N2) and tm3603 animals. (B) Maximum projection of Z-stack images at days 1 and 5 of adulthood showing YFP fluorescent puncta. (C) Number of puncta of diameter 2 μm and above determined by manual counting and automated analysis of total pixel area after thresholding. Experiments performed in triplicate ± SEMs, * p

    Techniques Used: Expressing

    MANF and CDNF homologs in selected organisms. (A) Protein domains and structure of the C. elegans manf-1 protein. (B) Schematic representation of MANF and CDNF proteins from Caenorhabditis elegans (Ce), Drosophila melanogaster ( Dm ), Homo sapiens ( Hs ) and Mus musculus ( Mm ) with percent identity and similarity indicated relative to C. elegans manf-1 . Conserved domains are aligned and signal sequences (SS) are depicted with sizes, all presented to scale. GenBank accession numbers for amino acid sequences are described in Figure S1 .
    Figure Legend Snippet: MANF and CDNF homologs in selected organisms. (A) Protein domains and structure of the C. elegans manf-1 protein. (B) Schematic representation of MANF and CDNF proteins from Caenorhabditis elegans (Ce), Drosophila melanogaster ( Dm ), Homo sapiens ( Hs ) and Mus musculus ( Mm ) with percent identity and similarity indicated relative to C. elegans manf-1 . Conserved domains are aligned and signal sequences (SS) are depicted with sizes, all presented to scale. GenBank accession numbers for amino acid sequences are described in Figure S1 .

    Techniques Used:

    4) Product Images from "Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells"

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173035

    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P
    Figure Legend Snippet: Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Techniques Used: Expressing, Western Blot

    Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.
    Figure Legend Snippet: Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Techniques Used: Activation Assay

    Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P
    Figure Legend Snippet: Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Techniques Used: Expressing, Binding Assay, Western Blot

    5) Product Images from "PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro"

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174968

    Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used:

    Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used: Activity Assay, Software

    6) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    7) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    8) Product Images from "Electrical synapses interconnecting axons revealed in the optic nerve head – a novel model of gap junctions’ involvement in optic nerve function"

    Article Title: Electrical synapses interconnecting axons revealed in the optic nerve head – a novel model of gap junctions’ involvement in optic nerve function

    Journal: Acta Ophthalmologica

    doi: 10.1111/aos.14272

    Expression and localization of connexin 36 and 45 in rat and human retina and optic nerve. (A–F) immunofluorescence staining of rat, whole‐mounted retinas for Cx36 (A–C) and Cx45 (D–F) in colocalization with RGC marker, Tuij1 ( β 3tubulin); nuclei are counterstained with DAPI . Scale bar, 10 μ m. In these whole‐mounted retina photographs, Cx36 expression within RGC s was present mostly intracellularly, while Cx45 showed a cell membrane pattern. In ON longitudinal sections, staining for both markers (Cx36 and Cx45) was observed along Tuij1‐positive axons. (G, H) immunofluorescence staining of paraffin longitudinal sections of a rat optic nerve head region for Cx36 (G) and Cx45 (H) in colocalization with RGC marker, Tuij1 ( β 3tubulin). Scale bar, 10 μ m. (I–N) immunofluorescence staining of longitudinal paraffin sections of a human optic nerve head region for Cx36 (I, J) and Cx45 (K, L) in colocalization with RGC marker, Tuij1 ( β 3tubulin), and Cx36 (N) and Cx45 (M) in colocalization with glial cell marker, GFAP . Scale bars, 10 μ m (I, K, N, M); 2 μ m (J, L).
    Figure Legend Snippet: Expression and localization of connexin 36 and 45 in rat and human retina and optic nerve. (A–F) immunofluorescence staining of rat, whole‐mounted retinas for Cx36 (A–C) and Cx45 (D–F) in colocalization with RGC marker, Tuij1 ( β 3tubulin); nuclei are counterstained with DAPI . Scale bar, 10 μ m. In these whole‐mounted retina photographs, Cx36 expression within RGC s was present mostly intracellularly, while Cx45 showed a cell membrane pattern. In ON longitudinal sections, staining for both markers (Cx36 and Cx45) was observed along Tuij1‐positive axons. (G, H) immunofluorescence staining of paraffin longitudinal sections of a rat optic nerve head region for Cx36 (G) and Cx45 (H) in colocalization with RGC marker, Tuij1 ( β 3tubulin). Scale bar, 10 μ m. (I–N) immunofluorescence staining of longitudinal paraffin sections of a human optic nerve head region for Cx36 (I, J) and Cx45 (K, L) in colocalization with RGC marker, Tuij1 ( β 3tubulin), and Cx36 (N) and Cx45 (M) in colocalization with glial cell marker, GFAP . Scale bars, 10 μ m (I, K, N, M); 2 μ m (J, L).

    Techniques Used: Expressing, Immunofluorescence, Staining, Marker

    Molecular analysis of Cx 36 and Cx 45 transcript and protein in retina and ON homogenates. (A) representative Western blot of homogenates prepared from rat retina and proximal and distal optic nerve stumps to detect the presence of Cx36 and Cx45. For the WB analysis, 2 retinas or 2 optic nerve stumps were pooled to obtain proper protein content. ON = optic nerve. (B) representative PCR of homogenates prepared from rat retina and proximal and distal optic nerve stumps to detect the presence of Cx36 and Cx45 transcripts. For the PCR analysis, 2 retinas or 2 optic nerve stumps were pooled to obtain proper RNA concentration. Rat brain cortex was used as a positive control and RNA ‐free water as a negative control. ON = optic nerve.
    Figure Legend Snippet: Molecular analysis of Cx 36 and Cx 45 transcript and protein in retina and ON homogenates. (A) representative Western blot of homogenates prepared from rat retina and proximal and distal optic nerve stumps to detect the presence of Cx36 and Cx45. For the WB analysis, 2 retinas or 2 optic nerve stumps were pooled to obtain proper protein content. ON = optic nerve. (B) representative PCR of homogenates prepared from rat retina and proximal and distal optic nerve stumps to detect the presence of Cx36 and Cx45 transcripts. For the PCR analysis, 2 retinas or 2 optic nerve stumps were pooled to obtain proper RNA concentration. Rat brain cortex was used as a positive control and RNA ‐free water as a negative control. ON = optic nerve.

    Techniques Used: Western Blot, Polymerase Chain Reaction, Concentration Assay, Positive Control, Negative Control

    Immunofluorescence staining of paraffin longitudinal sections of the ON from two healthy human donors. (A–D) immunostaining for Cx36 and Tuij1 showed dots of Cx36 staining colocalized with Tuij1‐positive ON axons. (E–H) immunostaining for Cx45 and Tuij1 showed dots of Cx45 staining colocalized with Tuij1‐positive ON axons. (I, J) photographs show Z‐stack imaging of longitudinal paraffin sections of the human optic nerve head region stained for Cx45 and Tuij1, with arrows pointing to the colocalization of axons with Cx45 projected in two planes. Scale bars, 10 μ m (A, B, E, F, I, J); 5 μ m (C, D, G, H).
    Figure Legend Snippet: Immunofluorescence staining of paraffin longitudinal sections of the ON from two healthy human donors. (A–D) immunostaining for Cx36 and Tuij1 showed dots of Cx36 staining colocalized with Tuij1‐positive ON axons. (E–H) immunostaining for Cx45 and Tuij1 showed dots of Cx45 staining colocalized with Tuij1‐positive ON axons. (I, J) photographs show Z‐stack imaging of longitudinal paraffin sections of the human optic nerve head region stained for Cx45 and Tuij1, with arrows pointing to the colocalization of axons with Cx45 projected in two planes. Scale bars, 10 μ m (A, B, E, F, I, J); 5 μ m (C, D, G, H).

    Techniques Used: Immunofluorescence, Staining, Immunostaining, Imaging

    9) Product Images from "PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro"

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174968

    PEDF down-regulates CD47 in vitro . ( A ) Total RNAs from PC3-Ctrl, PC3-PEDF, CL1-Ctrl, and CL1-PEDF were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to control cells. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p
    Figure Legend Snippet: PEDF down-regulates CD47 in vitro . ( A ) Total RNAs from PC3-Ctrl, PC3-PEDF, CL1-Ctrl, and CL1-PEDF were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to control cells. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p

    Techniques Used: In Vitro, Quantitative RT-PCR

    PEDF receptors expression in vitro . ( A ) Total RNAs from RAW 264.7 and BMDMs (tumor-free mice) were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to ATP5B mRNA expression levels in macrophages. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p
    Figure Legend Snippet: PEDF receptors expression in vitro . ( A ) Total RNAs from RAW 264.7 and BMDMs (tumor-free mice) were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to ATP5B mRNA expression levels in macrophages. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p

    Techniques Used: Expressing, In Vitro, Mouse Assay, Quantitative RT-PCR

    10) Product Images from "Characterization of the small molecule ARC39, a direct and specific inhibitor of acid sphingomyelinase in vitro [S]"

    Article Title: Characterization of the small molecule ARC39, a direct and specific inhibitor of acid sphingomyelinase in vitro [S]

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.RA120000682

    Direct and specific inhibition of ASM by ARC39 under cell culture conditions. A: L929 cells were treated for 2 h with ARC39 as indicated, and the activity of ASM, NSM, and NC was determined. L929 cells were treated for 2 h with ARC39, amitriptyline (Ami) or desipramine (Desi) as indicated, and AC activity was subsequently determined in B, and AC protein was analyzed by Western blotting in C. D: L929 cells were treated with 20 μM of ARC39 as indicated: fold change of Smpd1 mRNA relative to Hprt1 (left), ASM protein level (representative Western blot) (right). E: ASM activity in L929 cells after treatment with ARC39 as indicated. P
    Figure Legend Snippet: Direct and specific inhibition of ASM by ARC39 under cell culture conditions. A: L929 cells were treated for 2 h with ARC39 as indicated, and the activity of ASM, NSM, and NC was determined. L929 cells were treated for 2 h with ARC39, amitriptyline (Ami) or desipramine (Desi) as indicated, and AC activity was subsequently determined in B, and AC protein was analyzed by Western blotting in C. D: L929 cells were treated with 20 μM of ARC39 as indicated: fold change of Smpd1 mRNA relative to Hprt1 (left), ASM protein level (representative Western blot) (right). E: ASM activity in L929 cells after treatment with ARC39 as indicated. P

    Techniques Used: Inhibition, Cell Culture, Activity Assay, Western Blot

    11) Product Images from "Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats"

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00380.2014

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,
    Figure Legend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Techniques Used: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric
    Figure Legend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Techniques Used: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P
    Figure Legend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Techniques Used: Expressing

    12) Product Images from "PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro"

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174968

    Effect of Atglistatin on macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with PEDF (10 nM), Atglistatin (40 μM) or PEDF/Atglistatin combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Effect of Atglistatin on macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with PEDF (10 nM), Atglistatin (40 μM) or PEDF/Atglistatin combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used: Activity Assay, Software

    Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used:

    PEDF receptors expression  in vitro . ( A ) Total RNAs from RAW 264.7 and BMDMs (tumor-free mice) were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to ATP5B mRNA expression levels in macrophages. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p
    Figure Legend Snippet: PEDF receptors expression in vitro . ( A ) Total RNAs from RAW 264.7 and BMDMs (tumor-free mice) were analyzed by qRT-PCR and normalized to S15. Results are presented as relative fold change compared to ATP5B mRNA expression levels in macrophages. Data points represent mean ± SD of triplicate samples from two independent experiments. Statistical analyses were performed using the Student’s t test, *: p

    Techniques Used: Expressing, In Vitro, Mouse Assay, Quantitative RT-PCR

    PEDF induces the migration of RAW 264.7 macrophages toward 3D CL1 tumor spheroids  in vitro . ( A ) Migration of activated RAW 264.7 macrophages (Green) assessed by confocal microscopy, towards 3D CL1-Ctrl tumor spheroids (Red) ± 10 nM PEDF for 24 hours. ( B ) Quantitative analyses using the two point analysis function (NIS-Elements AR 4.00.03, Carl Zeiss) and showing that PEDF stimulates significantly the migration of macrophages towards the spheroids. Data points represent mean ± SD of triplicate samples from three independent experiments (*p
    Figure Legend Snippet: PEDF induces the migration of RAW 264.7 macrophages toward 3D CL1 tumor spheroids in vitro . ( A ) Migration of activated RAW 264.7 macrophages (Green) assessed by confocal microscopy, towards 3D CL1-Ctrl tumor spheroids (Red) ± 10 nM PEDF for 24 hours. ( B ) Quantitative analyses using the two point analysis function (NIS-Elements AR 4.00.03, Carl Zeiss) and showing that PEDF stimulates significantly the migration of macrophages towards the spheroids. Data points represent mean ± SD of triplicate samples from three independent experiments (*p

    Techniques Used: Migration, In Vitro, Confocal Microscopy

    PEDF increases the phagocytosis of PCa cells by RAW 264.7 macrophages. ( A, Left ) Representative fluorescent and Nomarski images of PC3-Ctrl or CL1-Ctrl PCa cells (Red) co-cultured with RAW 264.7 macrophages ± PEDF (10 nM). Thin arrows denote macrophages differentiation. Thick arrows show tumor cell debris (Red) within the macrophages. Nomarski and confocal images were obtained using the Nikon T1-E microscope with A1 confocal and STORM super-resolution with a 63x objective (N.A. 1.4; oil). After imaging, Regions of Interest (ROIs) were selected, and the intensity surface plot function (NIS-Elements AR 4.00.03) was used to measure the signal intensity of each ROI. ROI mean intensity was calculated from  > 30 representative ROI.  Right:  Quantification of tumor cell phagocytosis. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from three independent experiments. Data were represented using a boxplot graph (SPSS 23 software for Windows) showing the median, inter-quartile range, upper and lower quartiles, and whiskers. Statistical analysis was performed using the Student's t test. *: P
    Figure Legend Snippet: PEDF increases the phagocytosis of PCa cells by RAW 264.7 macrophages. ( A, Left ) Representative fluorescent and Nomarski images of PC3-Ctrl or CL1-Ctrl PCa cells (Red) co-cultured with RAW 264.7 macrophages ± PEDF (10 nM). Thin arrows denote macrophages differentiation. Thick arrows show tumor cell debris (Red) within the macrophages. Nomarski and confocal images were obtained using the Nikon T1-E microscope with A1 confocal and STORM super-resolution with a 63x objective (N.A. 1.4; oil). After imaging, Regions of Interest (ROIs) were selected, and the intensity surface plot function (NIS-Elements AR 4.00.03) was used to measure the signal intensity of each ROI. ROI mean intensity was calculated from > 30 representative ROI. Right: Quantification of tumor cell phagocytosis. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from three independent experiments. Data were represented using a boxplot graph (SPSS 23 software for Windows) showing the median, inter-quartile range, upper and lower quartiles, and whiskers. Statistical analysis was performed using the Student's t test. *: P

    Techniques Used: Cell Culture, Microscopy, Imaging, Software

    Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used: Activity Assay, Software

    PEDF induces the phagocytosis of tumor cells through an apoptosis-dependent mechanism and possibly via the production of Superoxide radicals. ( A, Left ) Representative images of CL1-Ctrl PCa cells (Red) and RAW 264.7 macrophages treated with buffer control, PEDF (10 nM), ZVAD (40 μM), or combination. ( A, Right ) Quantification of tumor cell phagocytosis (ROI mean intensity) averaged from two different experiments. Data were represented using a boxplot graph showing the median, inter-quartile range, upper and lower quartiles, and whiskers. Statistical analysis was performed using the Student's t test. *: P
    Figure Legend Snippet: PEDF induces the phagocytosis of tumor cells through an apoptosis-dependent mechanism and possibly via the production of Superoxide radicals. ( A, Left ) Representative images of CL1-Ctrl PCa cells (Red) and RAW 264.7 macrophages treated with buffer control, PEDF (10 nM), ZVAD (40 μM), or combination. ( A, Right ) Quantification of tumor cell phagocytosis (ROI mean intensity) averaged from two different experiments. Data were represented using a boxplot graph showing the median, inter-quartile range, upper and lower quartiles, and whiskers. Statistical analysis was performed using the Student's t test. *: P

    Techniques Used:

    13) Product Images from "CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target "

    Article Title: CAIX furthers tumour progression in the hypoxic tumour microenvironment of esophageal carcinoma and is a possible therapeutic target

    Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

    doi: 10.1080/14756366.2018.1475369

    Esophageal carcinoma tissue and microarray. (A) Tissue micro array (TMA) of esophageal carcinoma primary tumour and metastatic tissue was examined for CAIX expression and grouped into negative, moderate, and strong CAIX expression. Figure 1(A) shows representative samples of immunohistochemical CAIX staining for adenocarcinoma and squamous cell carcinoma at a standard magnification of ×20. (B ) Patients with a moderate CAIX expression had a significantly worse overall ( p = .003 and p = .009) and disease free ( p = .006 and p = .015) survival compared to patients with no or a strong CAIX expression. (C,D ) CAIX expression of primary tumour was correlated with CAIX expression of lymph node and organ metastases (representative immunohistochemical stainings are shown in Figure 1(C). Figure 1(D) shows the correlation of the intensity of CAIX staining (0-none, 1-low, 2-moderate, 3-strong, 4-strongest) of primary tumour and metastases. The left graph depicts the intensity of CAIX staining of the lymph node metastases, the right graph shows the intensities of the organ metastases in correlation to the intensity score of their primary tumour. Interestingly, most lymph node metastases occurred in the group with a moderate CAIX expression and thus impaired prognosis (group 2 in Figure 1(D)) and the metastatic lymph nodes of these patients overall expressed CAIX in higher intensities than any of the others. (E) All tumour samples ( n = 10), although to a different extent, showed an increased expression of RNA levels in quantitative PCR compared to their respective normal mucosal control tissues.
    Figure Legend Snippet: Esophageal carcinoma tissue and microarray. (A) Tissue micro array (TMA) of esophageal carcinoma primary tumour and metastatic tissue was examined for CAIX expression and grouped into negative, moderate, and strong CAIX expression. Figure 1(A) shows representative samples of immunohistochemical CAIX staining for adenocarcinoma and squamous cell carcinoma at a standard magnification of ×20. (B ) Patients with a moderate CAIX expression had a significantly worse overall ( p = .003 and p = .009) and disease free ( p = .006 and p = .015) survival compared to patients with no or a strong CAIX expression. (C,D ) CAIX expression of primary tumour was correlated with CAIX expression of lymph node and organ metastases (representative immunohistochemical stainings are shown in Figure 1(C). Figure 1(D) shows the correlation of the intensity of CAIX staining (0-none, 1-low, 2-moderate, 3-strong, 4-strongest) of primary tumour and metastases. The left graph depicts the intensity of CAIX staining of the lymph node metastases, the right graph shows the intensities of the organ metastases in correlation to the intensity score of their primary tumour. Interestingly, most lymph node metastases occurred in the group with a moderate CAIX expression and thus impaired prognosis (group 2 in Figure 1(D)) and the metastatic lymph nodes of these patients overall expressed CAIX in higher intensities than any of the others. (E) All tumour samples ( n = 10), although to a different extent, showed an increased expression of RNA levels in quantitative PCR compared to their respective normal mucosal control tissues.

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

    14) Product Images from "PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro"

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174968

    Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used:

    Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p
    Figure Legend Snippet: Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Techniques Used: Activity Assay, Software

    15) Product Images from "Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells"

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0173035

    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P
    Figure Legend Snippet: Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Techniques Used: Expressing, Western Blot

    Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.
    Figure Legend Snippet: Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Techniques Used: Activation Assay

    Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P
    Figure Legend Snippet: Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Techniques Used: Expressing, Binding Assay, Western Blot

    16) Product Images from "HSF1 deficiency and impaired HSP90-dependent protein folding are hallmarks of aneuploid human cells"

    Article Title: HSF1 deficiency and impaired HSP90-dependent protein folding are hallmarks of aneuploid human cells

    Journal: The EMBO Journal

    doi: 10.15252/embj.201488648

    The basal and stress-induced activity of HSF1 is impaired in human aneuploid cells A, B Western blot analysis for HSP27, HSP70, HSP90 (the used antibody recognizes both constitutive and inducible forms of HSP90) and HSF1 in parental and aneuploid cell lines (A). Loading control: GAPDH; HSC70 (constitutively expressed chaperone) in RPE-1 5/3 12/3 and corresponding control (note that GAPDH is encoded on chromosome 12). Shown are representative images of at least 3 independent experiments. In panel B the quantification of the signal intensities from the Western blots shown in (A) are depicted, calculated relative to control cells (which were set to 1). C, D HSP70-luc plasmid was expressed in parental and aneuploid HCT116 and RPE-1 cell lines for 36 h. Cells were then incubated with solvent control (DMSO), 2 μM 17-AAG or 5 μM MG132 for the indicated times. The depicted values show the fold induction in 17-AAG- or MG132-treated cells compared to DMSO-treated cells (which were set to 1). E HCT116 (left panel) and RPE-1 (right panel) cells were transfected with siRNA targeting HSF1 or the GL2 subunit of luciferase as a control (ctrl). Cell extract was prepared 72 h after transfection and subjected to immunoblotting for HSF1 and GAPDH as a loading control. Quantification of the signal normalized to the loading control is shown above the images. F HCT116 (left panel) and RPE-1 (right panel) cells transfected with siRNA targeting HSF1 or the GL2 subunit of luciferase as a control (ctrl). Forty-eight hours after transfection cells were incubated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. Data information: All data are the mean of at least three independent experiments ± SEM. * P
    Figure Legend Snippet: The basal and stress-induced activity of HSF1 is impaired in human aneuploid cells A, B Western blot analysis for HSP27, HSP70, HSP90 (the used antibody recognizes both constitutive and inducible forms of HSP90) and HSF1 in parental and aneuploid cell lines (A). Loading control: GAPDH; HSC70 (constitutively expressed chaperone) in RPE-1 5/3 12/3 and corresponding control (note that GAPDH is encoded on chromosome 12). Shown are representative images of at least 3 independent experiments. In panel B the quantification of the signal intensities from the Western blots shown in (A) are depicted, calculated relative to control cells (which were set to 1). C, D HSP70-luc plasmid was expressed in parental and aneuploid HCT116 and RPE-1 cell lines for 36 h. Cells were then incubated with solvent control (DMSO), 2 μM 17-AAG or 5 μM MG132 for the indicated times. The depicted values show the fold induction in 17-AAG- or MG132-treated cells compared to DMSO-treated cells (which were set to 1). E HCT116 (left panel) and RPE-1 (right panel) cells were transfected with siRNA targeting HSF1 or the GL2 subunit of luciferase as a control (ctrl). Cell extract was prepared 72 h after transfection and subjected to immunoblotting for HSF1 and GAPDH as a loading control. Quantification of the signal normalized to the loading control is shown above the images. F HCT116 (left panel) and RPE-1 (right panel) cells transfected with siRNA targeting HSF1 or the GL2 subunit of luciferase as a control (ctrl). Forty-eight hours after transfection cells were incubated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. Data information: All data are the mean of at least three independent experiments ± SEM. * P

    Techniques Used: Activity Assay, Western Blot, Plasmid Preparation, Incubation, Transfection, Luciferase

    Endogenous or exogenous overexpression of HSF1 ameliorates the negative effects of aneuploidy on protein folding A, B Western blot analysis of HSF1 and HSP90 expression in HCT116 8/3* c1-c4 (A). Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities from the Western blots (B), calculated relative to control cells (which were set to 1). C FlucDM-mCherry was expressed in parental HCT116* and HCT116* 8/3 c1-c4 for 36 h. Cells were then incubated with either solvent control (DMSO) or 50 nM 17-AAG for 8 h followed by measurement of luminescent activity. The depicted values show the percentage of luminescence in cells treated with 17-AAG relative to DMSO-treated cells (which were set to 100%). D Parental HCT116* and HCT116* cells trisomic for chromosome 8 (HCT116 8/3* c1-c4) were treated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control (which was set to 100%). E Western blot analysis of HSF1 expression and its downstream targets in the indicated aneuploid cells transfected with ca-HSF1. Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities normalized to the loading control is shown above the images. F RPE-1* 21/3 cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Forty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. G FlucDM-mCherry was co-expressed with either pCDNA or ca-HSF1 in the indicated cell lines for 36 h. Cells were then incubated with either solvent control (DMSO), 50 nM 17-AAG (HCT116), or 5 nM 17-AAG (RPE-1) for 8 h followed by measurement of luminescent activity. The depicted values show the percentage of luminescence in cells treated with 17-AAG relative to DMSO-treated cells (which were set to 100%). H Western blot analysis of HSF1 expression and its downstream targets in the indicated aneuploid cells transfected with ca-HSF1 using electroporation. Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities normalized to the loading control is shown above the images. I Control HCT116* cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Fourty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. J Control RPE-1* cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Fourty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. Data information: The data are the mean of at least three independent experiments ± SEM. * P
    Figure Legend Snippet: Endogenous or exogenous overexpression of HSF1 ameliorates the negative effects of aneuploidy on protein folding A, B Western blot analysis of HSF1 and HSP90 expression in HCT116 8/3* c1-c4 (A). Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities from the Western blots (B), calculated relative to control cells (which were set to 1). C FlucDM-mCherry was expressed in parental HCT116* and HCT116* 8/3 c1-c4 for 36 h. Cells were then incubated with either solvent control (DMSO) or 50 nM 17-AAG for 8 h followed by measurement of luminescent activity. The depicted values show the percentage of luminescence in cells treated with 17-AAG relative to DMSO-treated cells (which were set to 100%). D Parental HCT116* and HCT116* cells trisomic for chromosome 8 (HCT116 8/3* c1-c4) were treated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control (which was set to 100%). E Western blot analysis of HSF1 expression and its downstream targets in the indicated aneuploid cells transfected with ca-HSF1. Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities normalized to the loading control is shown above the images. F RPE-1* 21/3 cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Forty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG, and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. G FlucDM-mCherry was co-expressed with either pCDNA or ca-HSF1 in the indicated cell lines for 36 h. Cells were then incubated with either solvent control (DMSO), 50 nM 17-AAG (HCT116), or 5 nM 17-AAG (RPE-1) for 8 h followed by measurement of luminescent activity. The depicted values show the percentage of luminescence in cells treated with 17-AAG relative to DMSO-treated cells (which were set to 100%). H Western blot analysis of HSF1 expression and its downstream targets in the indicated aneuploid cells transfected with ca-HSF1 using electroporation. Loading control: GAPDH. Shown are representative images of at least 3 independent experiments. Quantification of the signal intensities normalized to the loading control is shown above the images. I Control HCT116* cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Fourty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. J Control RPE-1* cells were transiently transfected with either pCDNA or ca-HSF1 by electroporation. Fourty-eight hours post-transfection cells were incubated with the indicated concentrations of 17-AAG and cell number was determined 72 h thereafter. Cell number is shown as the percentage of the DMSO-treated control. Data information: The data are the mean of at least three independent experiments ± SEM. * P

    Techniques Used: Over Expression, Western Blot, Expressing, Incubation, Activity Assay, Transfection, Electroporation

    17) Product Images from "Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism"

    Article Title: Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-40482-0

    TNMD promotes adipogenesis and impairs lipid metabolism in human adipocytes. Human adipocytes were transfected with an adenovirus-5 containing a shRNA-TNMD and shRNA-control (scrambled) at day 14 of adipogenesis induction. (a–c ) Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer-binding protein alpha (CEBPA) and angiopoietin-like 4 (ANGPTL4) mRNA and protein levels were determined in the shRNA-TNMD and shRNA-control adipocytes. Protein levels in cell lysates were analyzed by Western blotting using specific antibodies against PPARG, CEBPA and ANGPTL4, normalized to the internal control (α-tubulin), and expressed as fold-change; the lower section shows a representative crop blot. (d – f) Hormone-sensitive lipase ( HSL ) gene expression, adipose triglyceride lipase ( ATGL ), and perilipin ( PLIN ) gene expression were determined in the shRNA-TNMD and shRNA-control adipocytes. ( g ) Glycerol levels (µM) in cell supernatants after treatment with shRNA-TNMD. All values are expressed as the means ± SEM of three independent experiments. Significant differences were identified using the nonparametric Mann-Whitney U test; *P-value
    Figure Legend Snippet: TNMD promotes adipogenesis and impairs lipid metabolism in human adipocytes. Human adipocytes were transfected with an adenovirus-5 containing a shRNA-TNMD and shRNA-control (scrambled) at day 14 of adipogenesis induction. (a–c ) Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer-binding protein alpha (CEBPA) and angiopoietin-like 4 (ANGPTL4) mRNA and protein levels were determined in the shRNA-TNMD and shRNA-control adipocytes. Protein levels in cell lysates were analyzed by Western blotting using specific antibodies against PPARG, CEBPA and ANGPTL4, normalized to the internal control (α-tubulin), and expressed as fold-change; the lower section shows a representative crop blot. (d – f) Hormone-sensitive lipase ( HSL ) gene expression, adipose triglyceride lipase ( ATGL ), and perilipin ( PLIN ) gene expression were determined in the shRNA-TNMD and shRNA-control adipocytes. ( g ) Glycerol levels (µM) in cell supernatants after treatment with shRNA-TNMD. All values are expressed as the means ± SEM of three independent experiments. Significant differences were identified using the nonparametric Mann-Whitney U test; *P-value

    Techniques Used: Transfection, shRNA, Binding Assay, Western Blot, Expressing, MANN-WHITNEY

    18) Product Images from "IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis"

    Article Title: IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.11.028

    Blockade of IL-33 signaling decreases chronic intestinal inflammation in SAMP1/YitFc (SAMP) mice. Disease severity ( A ) and representative images ( B ) of major basic protein–stained SAMP ilea show marked reduction in inflammation, improvement in villous structure, and overall gut morphology, and decreased infiltrating eosinophil using both preventive and therapeutic strategies after anti-ST2 administration compared with IgG-treated control (Cont.). Ileal eotaxin mRNA expression ( C ) and type 2 helper T-cell cytokine protein levels ( D ) from ex vivo T-cell receptor–activated mesenteric lymph node cells. ∗ P
    Figure Legend Snippet: Blockade of IL-33 signaling decreases chronic intestinal inflammation in SAMP1/YitFc (SAMP) mice. Disease severity ( A ) and representative images ( B ) of major basic protein–stained SAMP ilea show marked reduction in inflammation, improvement in villous structure, and overall gut morphology, and decreased infiltrating eosinophil using both preventive and therapeutic strategies after anti-ST2 administration compared with IgG-treated control (Cont.). Ileal eotaxin mRNA expression ( C ) and type 2 helper T-cell cytokine protein levels ( D ) from ex vivo T-cell receptor–activated mesenteric lymph node cells. ∗ P

    Techniques Used: Mouse Assay, Staining, Expressing, Ex Vivo

    Colonic IL-33, eosinophil (EOS) infiltration, and EOS-associated mediators are elevated in ulcerative colitis (UC) patients. Representative images of surgically resected, full-thickness colonic tissues stained for either IL-33 ( A and D ) or EOS protein X ( B and E ) show abundant IL-33 localized within the mucosa and submucosa of ulcerated, inflamed areas of UC patients ( A ) accompanied by marked accumulation of EOS distributed at the mucosal/submucosal interface in close proximity to IL-33 ( B ). D and E: Noninflamed areas of control (Cont.) patients show less IL-33, primarily limited to surface epithelial cells, and sparsely scattered EOS. C: IL5 and eotaxin-1 ( CCL11 ), eotaxin-2 ( CCL24 ), and eotaxin-3 ( CCL26 ) in involved and noninvolved colonic mucosal biopsy specimens from UC patients compared with Cont. ∗ P
    Figure Legend Snippet: Colonic IL-33, eosinophil (EOS) infiltration, and EOS-associated mediators are elevated in ulcerative colitis (UC) patients. Representative images of surgically resected, full-thickness colonic tissues stained for either IL-33 ( A and D ) or EOS protein X ( B and E ) show abundant IL-33 localized within the mucosa and submucosa of ulcerated, inflamed areas of UC patients ( A ) accompanied by marked accumulation of EOS distributed at the mucosal/submucosal interface in close proximity to IL-33 ( B ). D and E: Noninflamed areas of control (Cont.) patients show less IL-33, primarily limited to surface epithelial cells, and sparsely scattered EOS. C: IL5 and eotaxin-1 ( CCL11 ), eotaxin-2 ( CCL24 ), and eotaxin-3 ( CCL26 ) in involved and noninvolved colonic mucosal biopsy specimens from UC patients compared with Cont. ∗ P

    Techniques Used: Staining

    Exogenous administration of recombinant IL-33 (rIL-33) induces ileal eosinophil (EOS) infiltration and a potent type 2 helper T-cell (Th2) immune response in gut-associated lymphoid tissues from normal, uninflamed AKR/J (AKR) mice. A: Il33 in full-thickness ilea from SAMP1/YitFc (SAMP) and age-matched AKR and correlation with EOS number. B: Representative images of full-thickness AKR ilea show abundant infiltration of major basic protein–positive EOS after rIL-33 administration compared with vehicle-treated controls (Cont.). EOS counts and eotaxin mRNA levels in ilea ( C ) and Th2 cytokine protein levels from ex vivo T-cell receptor–activated mesenteric lymph node cells from IL-33– versus Cont.-treated mice ( D ). ∗ P
    Figure Legend Snippet: Exogenous administration of recombinant IL-33 (rIL-33) induces ileal eosinophil (EOS) infiltration and a potent type 2 helper T-cell (Th2) immune response in gut-associated lymphoid tissues from normal, uninflamed AKR/J (AKR) mice. A: Il33 in full-thickness ilea from SAMP1/YitFc (SAMP) and age-matched AKR and correlation with EOS number. B: Representative images of full-thickness AKR ilea show abundant infiltration of major basic protein–positive EOS after rIL-33 administration compared with vehicle-treated controls (Cont.). EOS counts and eotaxin mRNA levels in ilea ( C ) and Th2 cytokine protein levels from ex vivo T-cell receptor–activated mesenteric lymph node cells from IL-33– versus Cont.-treated mice ( D ). ∗ P

    Techniques Used: Recombinant, Mouse Assay, Ex Vivo

    Presence of commensal flora is essential for induction of IL-33, eosinophil (EOS) infiltration, and type 2 helper T-cell (Th2) immune responses in SAMP1/YitFc (SAMP) ileitis. Representative images ( A ) and quantitation of EOS ( B ) in MBP-stained full-thickness ilea show decreased infiltrating EOS in germ free (GF)-compared with specific pathogen-free (SPF) SAMP and GF-SAMP after fecal material transplantation (FMT). C: Il-33 in SPF-SAMP treated with combination anti-CCR3/IL-5 antibodies versus IgG-treated control (Cont.). D: Il-33 and Th2 cytokine mRNA expression in ilea of SPF- compared to age-matched GF-SAMP and GF-SAMP after fecal transplantation (FT). ∗ P
    Figure Legend Snippet: Presence of commensal flora is essential for induction of IL-33, eosinophil (EOS) infiltration, and type 2 helper T-cell (Th2) immune responses in SAMP1/YitFc (SAMP) ileitis. Representative images ( A ) and quantitation of EOS ( B ) in MBP-stained full-thickness ilea show decreased infiltrating EOS in germ free (GF)-compared with specific pathogen-free (SPF) SAMP and GF-SAMP after fecal material transplantation (FMT). C: Il-33 in SPF-SAMP treated with combination anti-CCR3/IL-5 antibodies versus IgG-treated control (Cont.). D: Il-33 and Th2 cytokine mRNA expression in ilea of SPF- compared to age-matched GF-SAMP and GF-SAMP after fecal transplantation (FT). ∗ P

    Techniques Used: Quantitation Assay, Staining, Transplantation Assay, Expressing

    19) Product Images from "Characterization of the small molecule ARC39, a direct and specific inhibitor of acid sphingomyelinase in vitro [S]"

    Article Title: Characterization of the small molecule ARC39, a direct and specific inhibitor of acid sphingomyelinase in vitro [S]

    Journal: Journal of Lipid Research

    doi: 10.1194/jlr.RA120000682

    Direct and specific inhibition of ASM by ARC39 under cell culture conditions. A: L929 cells were treated for 2 h with ARC39 as indicated, and the activity of ASM, NSM, and NC was determined. L929 cells were treated for 2 h with ARC39, amitriptyline (Ami) or desipramine (Desi) as indicated, and AC activity was subsequently determined in B, and AC protein was analyzed by Western blotting in C. D: L929 cells were treated with 20 μM of ARC39 as indicated: fold change of Smpd1 mRNA relative to Hprt1 (left), ASM protein level (representative Western blot) (right). E: ASM activity in L929 cells after treatment with ARC39 as indicated. P
    Figure Legend Snippet: Direct and specific inhibition of ASM by ARC39 under cell culture conditions. A: L929 cells were treated for 2 h with ARC39 as indicated, and the activity of ASM, NSM, and NC was determined. L929 cells were treated for 2 h with ARC39, amitriptyline (Ami) or desipramine (Desi) as indicated, and AC activity was subsequently determined in B, and AC protein was analyzed by Western blotting in C. D: L929 cells were treated with 20 μM of ARC39 as indicated: fold change of Smpd1 mRNA relative to Hprt1 (left), ASM protein level (representative Western blot) (right). E: ASM activity in L929 cells after treatment with ARC39 as indicated. P

    Techniques Used: Inhibition, Cell Culture, Activity Assay, Western Blot

    20) Product Images from "IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis"

    Article Title: IL-33 Drives Eosinophil Infiltration and Pathogenic Type 2 Helper T-Cell Immune Responses Leading to Chronic Experimental Ileitis

    Journal: The American Journal of Pathology

    doi: 10.1016/j.ajpath.2015.11.028

    Blockade of IL-33 signaling decreases chronic intestinal inflammation in SAMP1/YitFc (SAMP) mice. Disease severity ( A ) and representative images ( B ) of major basic protein–stained SAMP ilea show marked reduction in inflammation, improvement in villous structure, and overall gut morphology, and decreased infiltrating eosinophil using both preventive and therapeutic strategies after anti-ST2 administration compared with IgG-treated control (Cont.). Ileal eotaxin mRNA expression ( C ) and type 2 helper T-cell cytokine protein levels ( D ) from ex vivo T-cell receptor–activated mesenteric lymph node cells. ∗ P
    Figure Legend Snippet: Blockade of IL-33 signaling decreases chronic intestinal inflammation in SAMP1/YitFc (SAMP) mice. Disease severity ( A ) and representative images ( B ) of major basic protein–stained SAMP ilea show marked reduction in inflammation, improvement in villous structure, and overall gut morphology, and decreased infiltrating eosinophil using both preventive and therapeutic strategies after anti-ST2 administration compared with IgG-treated control (Cont.). Ileal eotaxin mRNA expression ( C ) and type 2 helper T-cell cytokine protein levels ( D ) from ex vivo T-cell receptor–activated mesenteric lymph node cells. ∗ P

    Techniques Used: Mouse Assay, Staining, Expressing, Ex Vivo

    Colonic IL-33, eosinophil (EOS) infiltration, and EOS-associated mediators are elevated in ulcerative colitis (UC) patients. Representative images of surgically resected, full-thickness colonic tissues stained for either IL-33 ( A and D ) or EOS protein X ( B and E ) show abundant IL-33 localized within the mucosa and submucosa of ulcerated, inflamed areas of UC patients ( A ) accompanied by marked accumulation of EOS distributed at the mucosal/submucosal interface in close proximity to IL-33 ( B ). D and E: Noninflamed areas of control (Cont.) patients show less IL-33, primarily limited to surface epithelial cells, and sparsely scattered EOS. C: IL5 and eotaxin-1 ( CCL11 ), eotaxin-2 ( CCL24 ), and eotaxin-3 ( CCL26 ) in involved and noninvolved colonic mucosal biopsy specimens from UC patients compared with Cont. ∗ P
    Figure Legend Snippet: Colonic IL-33, eosinophil (EOS) infiltration, and EOS-associated mediators are elevated in ulcerative colitis (UC) patients. Representative images of surgically resected, full-thickness colonic tissues stained for either IL-33 ( A and D ) or EOS protein X ( B and E ) show abundant IL-33 localized within the mucosa and submucosa of ulcerated, inflamed areas of UC patients ( A ) accompanied by marked accumulation of EOS distributed at the mucosal/submucosal interface in close proximity to IL-33 ( B ). D and E: Noninflamed areas of control (Cont.) patients show less IL-33, primarily limited to surface epithelial cells, and sparsely scattered EOS. C: IL5 and eotaxin-1 ( CCL11 ), eotaxin-2 ( CCL24 ), and eotaxin-3 ( CCL26 ) in involved and noninvolved colonic mucosal biopsy specimens from UC patients compared with Cont. ∗ P

    Techniques Used: Staining

    Exogenous administration of recombinant IL-33 (rIL-33) induces ileal eosinophil (EOS) infiltration and a potent type 2 helper T-cell (Th2) immune response in gut-associated lymphoid tissues from normal, uninflamed AKR/J (AKR) mice. A: Il33 in full-thickness ilea from SAMP1/YitFc (SAMP) and age-matched AKR and correlation with EOS number. B: Representative images of full-thickness AKR ilea show abundant infiltration of major basic protein–positive EOS after rIL-33 administration compared with vehicle-treated controls (Cont.). EOS counts and eotaxin mRNA levels in ilea ( C ) and Th2 cytokine protein levels from ex vivo T-cell receptor–activated mesenteric lymph node cells from IL-33– versus Cont.-treated mice ( D ). ∗ P
    Figure Legend Snippet: Exogenous administration of recombinant IL-33 (rIL-33) induces ileal eosinophil (EOS) infiltration and a potent type 2 helper T-cell (Th2) immune response in gut-associated lymphoid tissues from normal, uninflamed AKR/J (AKR) mice. A: Il33 in full-thickness ilea from SAMP1/YitFc (SAMP) and age-matched AKR and correlation with EOS number. B: Representative images of full-thickness AKR ilea show abundant infiltration of major basic protein–positive EOS after rIL-33 administration compared with vehicle-treated controls (Cont.). EOS counts and eotaxin mRNA levels in ilea ( C ) and Th2 cytokine protein levels from ex vivo T-cell receptor–activated mesenteric lymph node cells from IL-33– versus Cont.-treated mice ( D ). ∗ P

    Techniques Used: Recombinant, Mouse Assay, Ex Vivo

    Presence of commensal flora is essential for induction of IL-33, eosinophil (EOS) infiltration, and type 2 helper T-cell (Th2) immune responses in SAMP1/YitFc (SAMP) ileitis. Representative images ( A ) and quantitation of EOS ( B ) in MBP-stained full-thickness ilea show decreased infiltrating EOS in germ free (GF)-compared with specific pathogen-free (SPF) SAMP and GF-SAMP after fecal material transplantation (FMT). C: Il-33 in SPF-SAMP treated with combination anti-CCR3/IL-5 antibodies versus IgG-treated control (Cont.). D: Il-33 and Th2 cytokine mRNA expression in ilea of SPF- compared to age-matched GF-SAMP and GF-SAMP after fecal transplantation (FT). ∗ P
    Figure Legend Snippet: Presence of commensal flora is essential for induction of IL-33, eosinophil (EOS) infiltration, and type 2 helper T-cell (Th2) immune responses in SAMP1/YitFc (SAMP) ileitis. Representative images ( A ) and quantitation of EOS ( B ) in MBP-stained full-thickness ilea show decreased infiltrating EOS in germ free (GF)-compared with specific pathogen-free (SPF) SAMP and GF-SAMP after fecal material transplantation (FMT). C: Il-33 in SPF-SAMP treated with combination anti-CCR3/IL-5 antibodies versus IgG-treated control (Cont.). D: Il-33 and Th2 cytokine mRNA expression in ilea of SPF- compared to age-matched GF-SAMP and GF-SAMP after fecal transplantation (FT). ∗ P

    Techniques Used: Quantitation Assay, Staining, Transplantation Assay, Expressing

    21) Product Images from "Profiling the microRNA signature of the peripheral sensory ganglia in experimental autoimmune encephalomyelitis (EAE)"

    Article Title: Profiling the microRNA signature of the peripheral sensory ganglia in experimental autoimmune encephalomyelitis (EAE)

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-019-1600-7

    Validation of DEGs by qPCR. Bar graphs of a C3, b C5ar1, c Stat1, d Trem2, e Tlr7, and f Tlr8. * p
    Figure Legend Snippet: Validation of DEGs by qPCR. Bar graphs of a C3, b C5ar1, c Stat1, d Trem2, e Tlr7, and f Tlr8. * p

    Techniques Used: Real-time Polymerase Chain Reaction

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: HSF1 deficiency and impaired HSP90-dependent protein folding are hallmarks of aneuploid human cells
    Article Snippet: .. Total RNA was isolated using the RNeasy kit (Qiagen) and reverse-transcribed into cDNA using the First Strand cDNA synthesis kit (Roche). qPCR was performed with a HSF1 assay from Qiagen (Cat. No. 330001 PPH00164F) on a LightCycler 480 (Roche) instrument using the KAPA SYBR FAST master mix. ..

    Article Title: Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism
    Article Snippet: .. Next, we determined the differential gene expression levels of TNMD (330001 PHH12206A, Qiagen, Hilden, Germany), peroxisome proliferator-activated receptor gamma (PPARγ ), leptin (LEP ), and adiponectin (ADIPOQ ) during the adipogenic differentiation via qPCR using specific primer sequences (Table ). ..

    Isolation:

    Article Title: HSF1 deficiency and impaired HSP90-dependent protein folding are hallmarks of aneuploid human cells
    Article Snippet: .. Total RNA was isolated using the RNeasy kit (Qiagen) and reverse-transcribed into cDNA using the First Strand cDNA synthesis kit (Roche). qPCR was performed with a HSF1 assay from Qiagen (Cat. No. 330001 PPH00164F) on a LightCycler 480 (Roche) instrument using the KAPA SYBR FAST master mix. ..

    Quantitative RT-PCR:

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Article Snippet: .. Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15. ..

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats
    Article Snippet: .. RT-qPCR primer assays for GAPDH (330001 PPR06557A), NOS1 (330001 PPR44930E), and GPBAR1 (TGR5, 330001 PPR49442A) were purchased from QIAGEN (Valencia, CA). .. Fungizone antimycotic (15290-018) and penicillin-streptomycin (15070-063) were purchased from GIBCO, Life Sciences Solutions.

    Expressing:

    Article Title: Effects of X-chromosome Tenomodulin Genetic Variants on Obesity in a Children’s Cohort and Implications of the Gene in Adipocyte Metabolism
    Article Snippet: .. Next, we determined the differential gene expression levels of TNMD (330001 PHH12206A, Qiagen, Hilden, Germany), peroxisome proliferator-activated receptor gamma (PPARγ ), leptin (LEP ), and adiponectin (ADIPOQ ) during the adipogenic differentiation via qPCR using specific primer sequences (Table ). ..

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    Qiagen rt² qpcr primer assay
    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and <t>CYP26A1</t> transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P
    Rt² Qpcr Primer Assay, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Journal: PLoS ONE

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    doi: 10.1371/journal.pone.0173035

    Figure Lengend Snippet: Induction of RA metabolic enzymes in decidualizing cells. (a) Expression of DHRS3 , RDH12 , RBP4 and CYP26A1 transcripts in undifferentiated or cells decidualized with 8-bromo-cAMP, P4, and E in the presence or absence of 10 −8 M atRA or 10 −8 M atRald for 8 days (n = 9–11). The results show fold-change (mean ± SEM) in mRNA levels relative to expression levels in vehicle-treated undifferentiated HESCs ( dotted lines ). (b) Representative Western blot analysis of culture treated in parallel. β-actin served as a loading control. * P

    Article Snippet: We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Techniques: Expressing, Western Blot

    Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Journal: PLoS ONE

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    doi: 10.1371/journal.pone.0173035

    Figure Lengend Snippet: Schematic overview of the RA pathway. Retinol, hydrolyzed from retinyl ester, is converted to retinaldehyde (Rald) and generates retinoic acid (RA) following two-step oxidation. Intracellular RA binds to CRABP2 and activates heterodimer, RAR and RXR, leading to apoptotic machinery. By contrast, RA-dependent activation of nuclear receptor, PPAR β/δ is mediated by FABP5.

    Article Snippet: We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Techniques: Activation Assay

    Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Journal: PLoS ONE

    Article Title: Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

    doi: 10.1371/journal.pone.0173035

    Figure Lengend Snippet: Expression of cellular retinoic acid binding proteins in decidualizing HESCs. (a) CRABP2 and FABP5 transcript levels in undifferentiated or decidualized cells treated with 8-bromo-cAMP, P4, and E in combination with or without 10 −8 M atRA or 10 −8 M atRald for 8 days. The results show fold-change (mean ± SEM) relative to vehicle control ( dotted lines ). CRABP2 mRNA was measured in 8 primary HESC cultures and FABP5 mRNA in 9 cultures. (b) Representative Western blot analysis of retinoic acid binding proteins in whole cell lysates from undifferentiated or cells decidualized in the absence or presence of 10 −8 M atRA or 10 −8 M atRald. β-actin served as a loading control. E, cortisone; RA, retinoic acid; Rald, retinaldehyde. * indicates P

    Article Snippet: We used commercially available primers for FABP5 (330001 PPH02412E, QIAGEN, Mayland, USA) and Cyp26a1 (330001 PPH01229A, QIAGEN, Mayland, USA).

    Techniques: Expressing, Binding Assay, Western Blot

    Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Journal: PLoS ONE

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    doi: 10.1371/journal.pone.0174968

    Figure Lengend Snippet: Effect of α-CD47 and Angiostatin combination on the phagocytosis of CL1 cells. Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl) or Angiostatin (10 nM) alone or in combination. Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Article Snippet: Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Techniques:

    Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Journal: PLoS ONE

    Article Title: PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro

    doi: 10.1371/journal.pone.0174968

    Figure Lengend Snippet: Role of PNPLA2, ATP5B and CD47 in macrophages differentiation and phagocytic activity. ( A ) Quantification analyses of PCa cell phagocytosis in CL1-Ctrl/RAW 264.7 co-cultures treated with α-CD47 (100 ng/μl), PEDF or P18 (10 nM) alone or PEDF/P18 in combination with either S-BEL (5 μM) or Angiostatin (10 nM). Data points represent ROI mean intensity ± SD of triplicate samples per treatment condition from two independent experiments. ( B ) Quantification analyses of the «dendrite-like» structure length using the ImageJ software. Data points show the mean ± SD of triplicate samples per treatment condition from two independent experiments. Statistical analyses were performed using the Welch and Brown-Forsythe tests followed by the Games-Howell test, *: p

    Article Snippet: Total RNAs from RAW 264.7 cells treated with PEDF (10 nM), Angiostatin (10 nM), or combination were analyzed by qRT-PCR for Angiostatin receptors (Annexin A2 # 330001 PPM34399F, c-Met # 330001 PPM03726A, Integrin beta 3 # 330001 PPM03687E, and Integrin alpha V # 330001 PPM03662D; all from Qiagen) and normalized to S15.

    Techniques: Activity Assay, Software

    Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    doi: 10.1152/ajpgi.00380.2014

    Figure Lengend Snippet: Protein expression of nNOS and TGR5. Protein bands at 155 and 35 kDa that corresponded to the molecular weights of nNOS protein and TGR5 protein were detected in the gastric tissue from rats fed regular chow (CT) ( A ). High-fat feeding (high-fat diet,

    Article Snippet: RT-qPCR primer assays for GAPDH (330001 PPR06557A), NOS1 (330001 PPR44930E), and GPBAR1 (TGR5, 330001 PPR49442A) were purchased from QIAGEN (Valencia, CA).

    Techniques: Expressing

    Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    doi: 10.1152/ajpgi.00380.2014

    Figure Lengend Snippet: Activation of neuronal nitric oxide synthase (nNOS)- and TGR5-containing neurons in the gastric myenteric plexus following a 2-wk high-fat diet (HFD). A : nNOS- and TGR5-containing neurons were present in significant quantities in the gastric myenteric

    Article Snippet: RT-qPCR primer assays for GAPDH (330001 PPR06557A), NOS1 (330001 PPR44930E), and GPBAR1 (TGR5, 330001 PPR49442A) were purchased from QIAGEN (Valencia, CA).

    Techniques: Activation Assay

    Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Upregulation of bile acid receptor TGR5 and nNOS in gastric myenteric plexus is responsible for delayed gastric emptying after chronic high-fat feeding in rats

    doi: 10.1152/ajpgi.00380.2014

    Figure Lengend Snippet: Gene expression of nNOS and TGR5. Gene expression levels of nNOS and TGR5 increased in the gastric myenteric plexus in rats fed a high-fat diet (HFD); 67 and 111%, respectively ( n = 6, both ** P

    Article Snippet: RT-qPCR primer assays for GAPDH (330001 PPR06557A), NOS1 (330001 PPR44930E), and GPBAR1 (TGR5, 330001 PPR49442A) were purchased from QIAGEN (Valencia, CA).

    Techniques: Expressing