glyceraldehyde-3-phosphate dehydrogenase gapdh Millipore Search Results


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  • 99
    Millipore monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh 1 100 000 millipore
    Effects of ethanol and androgenization on profibrotic gene expression. Quantitative Real-Time PCR analysis of Interleukin-6 (IL-6) (black bar) and collagen Type 3 (COL3α1) (striped bar) gene expression in control (Ctrl), androgenized (Andro) and Andro + estrogen (Andro + E2) animals maintained on control Lieber-DeCarli (C-LDC) or ethanol-LDC (E-LDC) diet. Gene expression was normalized to <t>glyceraldehyde-3-phosphate</t> dehydrogenase <t>(GAPDH).</t> Data are presented as mean values ± SEM. *P
    Monoclonal Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh 1 100 000 Millipore, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh 1 100 000 millipore/product/Millipore
    Average 99 stars, based on 187 article reviews
    Price from $9.99 to $1999.99
    monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh 1 100 000 millipore - by Bioz Stars, 2020-08
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    91
    Merck KGaA glyceraldehyde 3 phosphate dehydrogenase gapdh
    Heat shock protein 70 (HSP70) expression and function manipulation affects Th17 gene expression. ( a ) Western blot analysis of HSP70 and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> proteins in the EL4 TCR + cells that were transfected with HSP70 siRNA or control in relation to the parallel molecular weight (MW) marker separation blot. Representative results from two independent experiments are shown. Gene expression analysis of the EL4 TCR + cells following anti-CD3 stimulation that has been transfected with HSP70 siRNA ( b ) or treated with pifithrin-μ (PFTμ) (10 μM) or with a control (dimethyl sulfoxide; DMSO) ( c ). ( d ) HSP70 gene expression changes of the EL4 TCR + cells following anti-CD3 stimulation or/and PFTμ (10 μM) treatment. Data represent mean +/− SD. * p
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glyceraldehyde 3 phosphate dehydrogenase gapdh/product/Merck KGaA
    Average 91 stars, based on 21 article reviews
    Price from $9.99 to $1999.99
    glyceraldehyde 3 phosphate dehydrogenase gapdh - by Bioz Stars, 2020-08
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    99
    Millipore human glyceraldehyde 3 phosphate dehydrogenase gapdh
    Validation of sortilin antibodies using sortilin knockout (−/−) and wildtype (+/+) mouse brains, with a characterization of the normal expression pattern of sortilin in rodent cerebrum. Panel (A) is a schematic drawing of the human sortilin protein, with the extracellular, transmembrane and intracellular domains of varying lengths of amino acid (a.a.) residues as marked. The immunogenic synthetic peptide sequences for the extracellular and intracellular C-terminal domain antibodies are also provided. Panels (B,C) show representative immunoblot and immunolabeling results obtained with the goat antibody (Gt Ab) against the extracellular domain in sortilin +/+ and −/− brains. Panels (D,E) show the results obtained with the rabbit antibody (Rt Ab) against the intracellular C-terminal. Both antibodies label a ~100 kDa band in sortilin +/+, but not in −/−, lysates (B,D) . Several non-specific bands are visible in the immunoblot with the goat antibody, equally present in the sortilin +/+ and −/− lysates, by extending the time of film exposure (B) . With overexposure, two light bands at ~40 and ~15 kDa (pointed by arrow) are also noticeable in the immunoblot of sortilin +/+ lysates with the rabbit antibody (D) . Light microscopic images (C,E) show neuronal profiles in the subiculum (Sub) to CA1 transitional region labeled by both antibodies in sortilin +/+, but not in −/−, brain sections. Confocal immunofluorescent images show completely colocalized labeling by the two antibodies in cortical pyramidal-like neurons (F–I) , CA3 pyramidal neurons and granule cells of the dentate gyrus (DG) (J–M) in C57BL mouse brain, with granular elements seen intracellularly (I,M) . Western blot applications: 12% SDS-PAGE gel and 13 μg equal amount protein loading. Additional abbreviations: CC, corpus callosum; s.p., stratum pyramidale; GCL, granule cell layer; PC, Parietal cortex; I-III, cortical layers; GAPHD, <t>glyceraldehyde-3-phosphate</t> dehydrogenase. Scale bar = 200 μm in (C) applying to (E) ; 100 μm in (F) applying to (G,H,J–L) , equivalent to 25 μm for (I,M) .
    Human Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 334 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human glyceraldehyde 3 phosphate dehydrogenase gapdh/product/Millipore
    Average 99 stars, based on 334 article reviews
    Price from $9.99 to $1999.99
    human glyceraldehyde 3 phosphate dehydrogenase gapdh - by Bioz Stars, 2020-08
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    99
    Millipore chloroplast gapdh
    Validation of sortilin antibodies using sortilin knockout (−/−) and wildtype (+/+) mouse brains, with a characterization of the normal expression pattern of sortilin in rodent cerebrum. Panel (A) is a schematic drawing of the human sortilin protein, with the extracellular, transmembrane and intracellular domains of varying lengths of amino acid (a.a.) residues as marked. The immunogenic synthetic peptide sequences for the extracellular and intracellular C-terminal domain antibodies are also provided. Panels (B,C) show representative immunoblot and immunolabeling results obtained with the goat antibody (Gt Ab) against the extracellular domain in sortilin +/+ and −/− brains. Panels (D,E) show the results obtained with the rabbit antibody (Rt Ab) against the intracellular C-terminal. Both antibodies label a ~100 kDa band in sortilin +/+, but not in −/−, lysates (B,D) . Several non-specific bands are visible in the immunoblot with the goat antibody, equally present in the sortilin +/+ and −/− lysates, by extending the time of film exposure (B) . With overexposure, two light bands at ~40 and ~15 kDa (pointed by arrow) are also noticeable in the immunoblot of sortilin +/+ lysates with the rabbit antibody (D) . Light microscopic images (C,E) show neuronal profiles in the subiculum (Sub) to CA1 transitional region labeled by both antibodies in sortilin +/+, but not in −/−, brain sections. Confocal immunofluorescent images show completely colocalized labeling by the two antibodies in cortical pyramidal-like neurons (F–I) , CA3 pyramidal neurons and granule cells of the dentate gyrus (DG) (J–M) in C57BL mouse brain, with granular elements seen intracellularly (I,M) . Western blot applications: 12% SDS-PAGE gel and 13 μg equal amount protein loading. Additional abbreviations: CC, corpus callosum; s.p., stratum pyramidale; GCL, granule cell layer; PC, Parietal cortex; I-III, cortical layers; GAPHD, <t>glyceraldehyde-3-phosphate</t> dehydrogenase. Scale bar = 200 μm in (C) applying to (E) ; 100 μm in (F) applying to (G,H,J–L) , equivalent to 25 μm for (I,M) .
    Chloroplast Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/chloroplast gapdh/product/Millipore
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    99
    Millipore glyceraldehyde 3 phosphate dehydrogenase gapdh monoclonal antibody
    Validation of sortilin antibodies using sortilin knockout (−/−) and wildtype (+/+) mouse brains, with a characterization of the normal expression pattern of sortilin in rodent cerebrum. Panel (A) is a schematic drawing of the human sortilin protein, with the extracellular, transmembrane and intracellular domains of varying lengths of amino acid (a.a.) residues as marked. The immunogenic synthetic peptide sequences for the extracellular and intracellular C-terminal domain antibodies are also provided. Panels (B,C) show representative immunoblot and immunolabeling results obtained with the goat antibody (Gt Ab) against the extracellular domain in sortilin +/+ and −/− brains. Panels (D,E) show the results obtained with the rabbit antibody (Rt Ab) against the intracellular C-terminal. Both antibodies label a ~100 kDa band in sortilin +/+, but not in −/−, lysates (B,D) . Several non-specific bands are visible in the immunoblot with the goat antibody, equally present in the sortilin +/+ and −/− lysates, by extending the time of film exposure (B) . With overexposure, two light bands at ~40 and ~15 kDa (pointed by arrow) are also noticeable in the immunoblot of sortilin +/+ lysates with the rabbit antibody (D) . Light microscopic images (C,E) show neuronal profiles in the subiculum (Sub) to CA1 transitional region labeled by both antibodies in sortilin +/+, but not in −/−, brain sections. Confocal immunofluorescent images show completely colocalized labeling by the two antibodies in cortical pyramidal-like neurons (F–I) , CA3 pyramidal neurons and granule cells of the dentate gyrus (DG) (J–M) in C57BL mouse brain, with granular elements seen intracellularly (I,M) . Western blot applications: 12% SDS-PAGE gel and 13 μg equal amount protein loading. Additional abbreviations: CC, corpus callosum; s.p., stratum pyramidale; GCL, granule cell layer; PC, Parietal cortex; I-III, cortical layers; GAPHD, <t>glyceraldehyde-3-phosphate</t> dehydrogenase. Scale bar = 200 μm in (C) applying to (E) ; 100 μm in (F) applying to (G,H,J–L) , equivalent to 25 μm for (I,M) .
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glyceraldehyde 3 phosphate dehydrogenase gapdh monoclonal antibody/product/Millipore
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    glyceraldehyde 3 phosphate dehydrogenase gapdh monoclonal antibody - by Bioz Stars, 2020-08
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    Image Search Results


    Effects of ethanol and androgenization on profibrotic gene expression. Quantitative Real-Time PCR analysis of Interleukin-6 (IL-6) (black bar) and collagen Type 3 (COL3α1) (striped bar) gene expression in control (Ctrl), androgenized (Andro) and Andro + estrogen (Andro + E2) animals maintained on control Lieber-DeCarli (C-LDC) or ethanol-LDC (E-LDC) diet. Gene expression was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data are presented as mean values ± SEM. *P

    Journal: PLoS ONE

    Article Title: Neonatal Androgenization Exacerbates Alcohol-Induced Liver Injury in Adult Rats, an Effect Abrogated by Estrogen

    doi: 10.1371/journal.pone.0029463

    Figure Lengend Snippet: Effects of ethanol and androgenization on profibrotic gene expression. Quantitative Real-Time PCR analysis of Interleukin-6 (IL-6) (black bar) and collagen Type 3 (COL3α1) (striped bar) gene expression in control (Ctrl), androgenized (Andro) and Andro + estrogen (Andro + E2) animals maintained on control Lieber-DeCarli (C-LDC) or ethanol-LDC (E-LDC) diet. Gene expression was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Data are presented as mean values ± SEM. *P

    Article Snippet: For Western blot analyses, antibodies against cytochrome P450 2E1 (CYP2E1), cytochrome P450 1A2 (CYP1A2) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    EibG expression under static growth conditions and with agitation. Cells of several STEC strains carrying the eib G gene were inoculated in LB medium with (+) and without (-) shaking at 37°C for 16h. Proteins were separated by SDS-PAGE and immunoblotted. (A, B) To compare expression levels, identical protein quantities of 7.5 μg were loaded in each lane. EibG was detected with human IgG Fc conjugated with HRP on immunoblots and visualized by chemiluminescence. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as marker protein and as control for loading. Marker sizes (M) and EibG proteins are indicated. (C) Proteins of strain 0520/99 were diluted serially as indicated and immunoblotted to demonstrate specifity and sensitivity to the detection platform. To standardize between immunoblots, the highest intensities were defined as 1.0 and the ratios of the diluted signals of three independent gel runs were calculated as means (± standard deviations of the means). Intensities of static grown bacteria and agitated cultures are shown by black and grey bars, respectively.

    Journal: PLoS ONE

    Article Title: Agitation Down-Regulates Immunoglobulin Binding Protein EibG Expression in Shiga Toxin-Producing Escherichia coli (STEC)

    doi: 10.1371/journal.pone.0119583

    Figure Lengend Snippet: EibG expression under static growth conditions and with agitation. Cells of several STEC strains carrying the eib G gene were inoculated in LB medium with (+) and without (-) shaking at 37°C for 16h. Proteins were separated by SDS-PAGE and immunoblotted. (A, B) To compare expression levels, identical protein quantities of 7.5 μg were loaded in each lane. EibG was detected with human IgG Fc conjugated with HRP on immunoblots and visualized by chemiluminescence. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as marker protein and as control for loading. Marker sizes (M) and EibG proteins are indicated. (C) Proteins of strain 0520/99 were diluted serially as indicated and immunoblotted to demonstrate specifity and sensitivity to the detection platform. To standardize between immunoblots, the highest intensities were defined as 1.0 and the ratios of the diluted signals of three independent gel runs were calculated as means (± standard deviations of the means). Intensities of static grown bacteria and agitated cultures are shown by black and grey bars, respectively.

    Article Snippet: Protein loading and regulation of EibG expression was controlled using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein as standard marker (Sigma-Aldrich, Germany) and the polyclonal goat anti-GAPDH antibody received as a purified IgG (antibodies-online, Germany).

    Techniques: Expressing, SDS Page, Western Blot, Marker

    uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm

    Journal: Fluids and Barriers of the CNS

    Article Title: Aquaporin 1 and the Na+/K+/2Cl− cotransporter 1 are present in the leptomeningeal vasculature of the adult rodent central nervous system

    doi: 10.1186/s12987-020-0176-z

    Figure Lengend Snippet: uDISCO clearance of the intact mouse head depicts the expression of aquaporin 1. a Mouse brain (P60) cleared by uDISCO and immunolabeled for AQP1 (AQP1int, green) reveals the vasculature network in the leptomeninges, including the middle cerebral arteries (MCA, arrows). AQP1 + cells also line the subarachnoid cisterns and the olfactory bulb. b Optical section reveals AQP1 + choroidal epithelial cells and olfactory ensheathing glia cells. c , d Higher magnification images of the areas depicted in b (blue and purple squares) showing AQP1 in the glomerular layer (arrow) and in choroidal epithelial cells (asterisk). e Representative micrograph of a parasagittal section of an adult mouse brain (P90) immunolabeled for AQP1 (AQP1ext, grey). AQP1ext + epithelial cells of the choroid plexus are observed in the fourth ( f ) and in the lateral ventricles ( g ). In contrast, olfactory ensheathing glia cells in the olfactory bulb are not immunolabeled ( h ). i Representative micrograph of a coronal section from adult mouse brain (P90) immunolabeled with AQP1 (AQP1int, grey). j Higher magnification of the depicted area in i (square) shows in detail AQP1int + epithelial cells in the choroid plexus of the lateral ventricles. k Olfactory ensheathing glia cells are also immunoreactive. Dashed line in k depicts the mitral cell layer. l Immunoblotting reveals a band of 35 kDa, corresponding to the glycosylated form of AQP1, detected in the BS, Cb, Ctx, Hip, Hyp and OB, obtained from young adult mice (P30). The non-glycosylated form of AQP1, corresponding to a band of 28 kDa, is detected in choroid plexi and kidney homogenates obtained from young adult mice (P30). The housekeeping protein GAPDH (37 kDa) was used as loading control. Control antigen confirms antibody-epitope specific binding. m Graphic shows the relative AQP1 protein levels, in relation to GAPDH. BS, brain stem; Cb, cerebellum; ChP, choroid plexus; Ctx, cerebral cortex; CPu, caudate putamen; EPL, external plexiform layer; Fi, fimbria; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GL, glomerular layer; Hip, hippocampus; Hyp, hypothalamus; IC, internal capsule; IPL, internal plexiform layer; Kdy, kidney; LV, lateral ventricle; OB, olfactory bulb; PirCtx, piriform cortex, SCh, suprachiasmatic nuclei; Thal, thalamus; WM, white matter; 3V, third ventricle; 4V, fourth ventricle. Scale bars: a , b , e 1 mm; c , i 500 μm; d , 200 μm; f – h , j , k 50 μm

    Article Snippet: The band corresponding to the housekeeping protein glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was detected by Western blot using a mouse anti-GAPDH antibody (Sigma-Aldrich, St. Louis, Missouri, USA), with an apparent molecular weight of 37 kDa, as previously described [ , ].

    Techniques: Expressing, Immunolabeling, Mouse Assay, Binding Assay

    Cardiac fatty acid metabolic proteins in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Representative immunoblots and average data of (a) ratio of phosphorylated 5′ adenosine monophosphate-activated protein kinase 2 α (pAMPK2 α ) to total AMPK2 α , (b) peroxisome proliferator-activated receptor- (PPAR-) γ coactivator- (PGC-) 1 α , (c) phosphorylated acetyl coenzyme A carboxylase (pACC), (d) cluster of differentiation 36 (CD36), (e) diacylglycerol acyltransferase 1 (DGAT1), and (f) DGAT2 from control ( n = 4), DM ( n = 4), and DM + MPT0E014 ( n = 4) rats. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Journal: PPAR Research

    Article Title: HDAC Inhibition Modulates Cardiac PPARs and Fatty Acid Metabolism in Diabetic Cardiomyopathy

    doi: 10.1155/2016/5938740

    Figure Lengend Snippet: Cardiac fatty acid metabolic proteins in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Representative immunoblots and average data of (a) ratio of phosphorylated 5′ adenosine monophosphate-activated protein kinase 2 α (pAMPK2 α ) to total AMPK2 α , (b) peroxisome proliferator-activated receptor- (PPAR-) γ coactivator- (PGC-) 1 α , (c) phosphorylated acetyl coenzyme A carboxylase (pACC), (d) cluster of differentiation 36 (CD36), (e) diacylglycerol acyltransferase 1 (DGAT1), and (f) DGAT2 from control ( n = 4), DM ( n = 4), and DM + MPT0E014 ( n = 4) rats. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Article Snippet: Targeted bands were normalized to cardiac glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Sigma-Aldrich) to confirm equal protein loading.

    Techniques: Western Blot, Pyrolysis Gas Chromatography

    Cardiac peroxisome proliferator-activated receptor (PPAR) proteins in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Cardiac PPAR- α and PPAR- δ protein expressions significantly decreased in DM ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Cardiac PPAR- γ protein expressions were enhanced in DM ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Representative immunoblots and average data of (a) PPAR- α , (b) PPAR- γ , and (c) PPAR- δ in different groups. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Journal: PPAR Research

    Article Title: HDAC Inhibition Modulates Cardiac PPARs and Fatty Acid Metabolism in Diabetic Cardiomyopathy

    doi: 10.1155/2016/5938740

    Figure Lengend Snippet: Cardiac peroxisome proliferator-activated receptor (PPAR) proteins in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Cardiac PPAR- α and PPAR- δ protein expressions significantly decreased in DM ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Cardiac PPAR- γ protein expressions were enhanced in DM ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Representative immunoblots and average data of (a) PPAR- α , (b) PPAR- γ , and (c) PPAR- δ in different groups. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Article Snippet: Targeted bands were normalized to cardiac glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Sigma-Aldrich) to confirm equal protein loading.

    Techniques: Western Blot

    Cardiac inflammatory proteins and ratio of phosphorylated Akt to total Akt in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Cardiac tumor necrosis factor- (TNF-) α and interleukin- (IL-) 6 protein levels were significantly higher in DM rats ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Representative immunoblots and average data of (a) TNF- α , (b) IL-6, and (c) ratio of pAkt to total Akt in different groups. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Journal: PPAR Research

    Article Title: HDAC Inhibition Modulates Cardiac PPARs and Fatty Acid Metabolism in Diabetic Cardiomyopathy

    doi: 10.1155/2016/5938740

    Figure Lengend Snippet: Cardiac inflammatory proteins and ratio of phosphorylated Akt to total Akt in control, diabetes mellitus (DM), and MPT0E014-treated DM (DM + MPT0E014) rats. Cardiac tumor necrosis factor- (TNF-) α and interleukin- (IL-) 6 protein levels were significantly higher in DM rats ( n = 4) compared to control ( n = 4) and DM + MPT0E014 ( n = 4) rats. Representative immunoblots and average data of (a) TNF- α , (b) IL-6, and (c) ratio of pAkt to total Akt in different groups. Densitometry was normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an internal control.

    Article Snippet: Targeted bands were normalized to cardiac glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Sigma-Aldrich) to confirm equal protein loading.

    Techniques: Western Blot

    Western blot results from rats receiving four treatments: PAH (pulmonary arterial hypertension), WGLL (Wild garlic ( Allium ursinum ) liophylisate-enriched chow), Sildenafil injection, and Control. ( A ) shows quantified results of Western blot analysis after normalization to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and data are presented as mean ± SEM for each treatment group. RV = right ventricle, L = lung ( n = 9; * p

    Journal: International Journal of Molecular Sciences

    Article Title: A Novel Therapeutic Approach in the Treatment of Pulmonary Arterial Hypertension: Allium ursinum Liophylisate Alleviates Symptoms Comparably to Sildenafil

    doi: 10.3390/ijms18071436

    Figure Lengend Snippet: Western blot results from rats receiving four treatments: PAH (pulmonary arterial hypertension), WGLL (Wild garlic ( Allium ursinum ) liophylisate-enriched chow), Sildenafil injection, and Control. ( A ) shows quantified results of Western blot analysis after normalization to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and data are presented as mean ± SEM for each treatment group. RV = right ventricle, L = lung ( n = 9; * p

    Article Snippet: After washing the membrane with TBS-T, primary PDE5A and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies (Sigma-Aldrich Co., St. Louis, MO, USA) diluted in TBS-T with 1% low-fat milk were added and incubated overnight at 4 °C.

    Techniques: Western Blot, Injection

    Validation of selective hit compounds to inhibit a CMV infection MRC5 cells pre-treated with DMSO, podofilox (5-50nM), convallatoxin (5-50nM), or colchicine (10-500nM) were infected with AD169-WT (MOI:3) and total cell lysates were resolved on a SDS-polyacrylamide gel (12%). Mock (lanes 1 and 12) and infected (lanes 2-11, 13-22) cells were harvested at 20 hpi and subjected to immunoblot analysis for the early CMV gene product US11 (lanes 1-11) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (lanes 12-22). The respective polypeptides and relative molecular mass markers are indicated.

    Journal: Antiviral research

    Article Title: Development of a high-content screen for the identification of inhibitors directed against the early steps of the cytomegalovirus infectious cycle

    doi: 10.1016/j.antiviral.2014.10.011

    Figure Lengend Snippet: Validation of selective hit compounds to inhibit a CMV infection MRC5 cells pre-treated with DMSO, podofilox (5-50nM), convallatoxin (5-50nM), or colchicine (10-500nM) were infected with AD169-WT (MOI:3) and total cell lysates were resolved on a SDS-polyacrylamide gel (12%). Mock (lanes 1 and 12) and infected (lanes 2-11, 13-22) cells were harvested at 20 hpi and subjected to immunoblot analysis for the early CMV gene product US11 (lanes 1-11) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (lanes 12-22). The respective polypeptides and relative molecular mass markers are indicated.

    Article Snippet: Total cell lysates from uninfected and infected cells 20 hpi were subjected to SDS-PAGE followed by immunoblot analysis using anti-CMV US11 antibody ( ) and an anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Chemicon, Billerica, MA).

    Techniques: Infection

    AD169 IE2-YFP -infected cells express IE2-YFP as an immediate-early protein (A) Mock (lanes 1, 5, and 9) and AD169 IE2-YFP -infected (lanes 2-4, 6-8, 10-12) (MOI:5) MRC5 cells were harvested up to 24 hpi and subjected to immunoblot analysis for CMV IE2 (lanes 1-4), CMV IE1 (lanes 5-8), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (lanes 9-12) polypeptides. The respective polypeptides and relative molecular mass markers are indicated. (B) Mock- and AD169 IE2-YFP -infected MRC5 cells (MOI:3) were stained with Hoechst reagent (blue) and analyzed by fluorescent microscopy (×20) at 24hpi. Both the Hoechst reagent and IE2-YFP (green) are localized to the nucleus and the overlay highlights the virus-infected cells.

    Journal: Antiviral research

    Article Title: Development of a high-content screen for the identification of inhibitors directed against the early steps of the cytomegalovirus infectious cycle

    doi: 10.1016/j.antiviral.2014.10.011

    Figure Lengend Snippet: AD169 IE2-YFP -infected cells express IE2-YFP as an immediate-early protein (A) Mock (lanes 1, 5, and 9) and AD169 IE2-YFP -infected (lanes 2-4, 6-8, 10-12) (MOI:5) MRC5 cells were harvested up to 24 hpi and subjected to immunoblot analysis for CMV IE2 (lanes 1-4), CMV IE1 (lanes 5-8), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (lanes 9-12) polypeptides. The respective polypeptides and relative molecular mass markers are indicated. (B) Mock- and AD169 IE2-YFP -infected MRC5 cells (MOI:3) were stained with Hoechst reagent (blue) and analyzed by fluorescent microscopy (×20) at 24hpi. Both the Hoechst reagent and IE2-YFP (green) are localized to the nucleus and the overlay highlights the virus-infected cells.

    Article Snippet: Total cell lysates from uninfected and infected cells 20 hpi were subjected to SDS-PAGE followed by immunoblot analysis using anti-CMV US11 antibody ( ) and an anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Chemicon, Billerica, MA).

    Techniques: Infection, Staining, Microscopy

    RACK1 was a directly target of miR-155. a Relative luciferase activity of recombinant RACK1 promotor and U6 (control) vectors in BV2 cells which were co-transfected with scramble control or miR-155 mimic ( n = 3). b Quantitative RT-PCR was performed to measure relative mRNA expression of RACK1 ( n = 3), and ( c ) Western blot analysis was performed to measure the protein expressions of RACK1 in cells which were transfected with scramble, miR-155 mimic, inhibitor control, and miR-155 inhibitor. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; RT-PCR: reverse transcription polymerase chain reaction. * P

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: RACK1 was a directly target of miR-155. a Relative luciferase activity of recombinant RACK1 promotor and U6 (control) vectors in BV2 cells which were co-transfected with scramble control or miR-155 mimic ( n = 3). b Quantitative RT-PCR was performed to measure relative mRNA expression of RACK1 ( n = 3), and ( c ) Western blot analysis was performed to measure the protein expressions of RACK1 in cells which were transfected with scramble, miR-155 mimic, inhibitor control, and miR-155 inhibitor. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; RT-PCR: reverse transcription polymerase chain reaction. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: Luciferase, Activity Assay, Recombinant, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

    RACK1 knockdown promoted LPS-induced cell injury. BV2 cells were treated with LPS, LPS+ inhibitor control, LPS + miR-155 inhibitor, and LPS+ miR-155 inhibitor + si-RACK1. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α were measured by quantitative RT-PCR ( n = 3). e - h Concentrations of IL-1β, IL-6, IL-8, and TNF-α were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: RACK1 knockdown promoted LPS-induced cell injury. BV2 cells were treated with LPS, LPS+ inhibitor control, LPS + miR-155 inhibitor, and LPS+ miR-155 inhibitor + si-RACK1. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α were measured by quantitative RT-PCR ( n = 3). e - h Concentrations of IL-1β, IL-6, IL-8, and TNF-α were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: CCK-8 Assay, Flow Cytometry, Cytometry, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Counting, Reverse Transcription Polymerase Chain Reaction

    Down-regulation of miR-155 protected BV2 cells from LPS-induced inflammatory injury via deactivation of MAPK/NF-κB and mTOR pathways. Western blot analysis was used to measure the expressions of RACK1, MAPK protein (p38), NF-κB proteins (p65 and lκBα), and mTOR proteins (mTOR and p70S6K) in BV2 cells which were treated with LPS, LPS + scramble, LPS + miR-155 mimic, LPS+ inhibitor control, and LPS + miR-155 inhibitor. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; LPS: lipopolysaccharide; MAPK: mitogen activated protein kinase; NF-κB: nuclear factor kappa B; mTOR: mammalian target of rapamycin

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: Down-regulation of miR-155 protected BV2 cells from LPS-induced inflammatory injury via deactivation of MAPK/NF-κB and mTOR pathways. Western blot analysis was used to measure the expressions of RACK1, MAPK protein (p38), NF-κB proteins (p65 and lκBα), and mTOR proteins (mTOR and p70S6K) in BV2 cells which were treated with LPS, LPS + scramble, LPS + miR-155 mimic, LPS+ inhibitor control, and LPS + miR-155 inhibitor. GAPDH: glyceraldehyde-3-phosphate dehydrogenase; LPS: lipopolysaccharide; MAPK: mitogen activated protein kinase; NF-κB: nuclear factor kappa B; mTOR: mammalian target of rapamycin

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: Western Blot

    LPS damaged mouse microglia BV2 cells in a time-dependent manner. BV2 cells were pre-treated with 10 μg/ml LPS for 0, 2, 4 and 5 h. Cells without LPS treatment were used as control. a Cell viability was measured by CCK-8 assay. b Cell apoptosis was measured by flow cytometry. c - f Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by quantitative RT-PCR. n = 3. CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: LPS damaged mouse microglia BV2 cells in a time-dependent manner. BV2 cells were pre-treated with 10 μg/ml LPS for 0, 2, 4 and 5 h. Cells without LPS treatment were used as control. a Cell viability was measured by CCK-8 assay. b Cell apoptosis was measured by flow cytometry. c - f Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by quantitative RT-PCR. n = 3. CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: CCK-8 Assay, Flow Cytometry, Cytometry, Quantitative RT-PCR, Cell Counting, Reverse Transcription Polymerase Chain Reaction

    LPS damaged mouse microglia BV2 cells in a dose-dependent manner. BV2 cells were pre-treated with different concentrations of LPS (0, 1, 5, 10, and 20 μg/ml) for 5 h. Cells without LPS treatment were used as control. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by quantitative RT-PCR ( n = 3). ( e - h ) Concentrations of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: LPS damaged mouse microglia BV2 cells in a dose-dependent manner. BV2 cells were pre-treated with different concentrations of LPS (0, 1, 5, 10, and 20 μg/ml) for 5 h. Cells without LPS treatment were used as control. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by quantitative RT-PCR ( n = 3). ( e - h ) Concentrations of IL-1β, IL-6, IL-8, and TNF-α in LPS-treated cells and control cells were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: CCK-8 Assay, Flow Cytometry, Cytometry, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Counting, Reverse Transcription Polymerase Chain Reaction

    Down-regulation of miR-155 inhibited LPS-induced cell injury. BV2 cells were administrated with LPS, LPS + scramble, LPS + miR-155 mimic, LPS+ inhibitor control, and LPS + miR-155 inhibitor. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α were measured by quantitative RT-PCR ( n = 3). e-h Concentrations of IL-1β, IL-6, IL-8, and TNF-α were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. ns, no significance, * P

    Journal: Journal of Inflammation (London, England)

    Article Title: Knockdown of miR-155 protects microglia against LPS-induced inflammatory injury via targeting RACK1: a novel research for intracranial infection

    doi: 10.1186/s12950-017-0162-7

    Figure Lengend Snippet: Down-regulation of miR-155 inhibited LPS-induced cell injury. BV2 cells were administrated with LPS, LPS + scramble, LPS + miR-155 mimic, LPS+ inhibitor control, and LPS + miR-155 inhibitor. a Cell viability was measured by CCK-8 assay ( n = 3). b Cell apoptosis was measured by flow cytometry ( n = 3). c Expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax, caspase-3, and caspase-9) proteins were measured by western blot analysis. d Relative mRNA expressions of IL-1β, IL-6, IL-8, and TNF-α were measured by quantitative RT-PCR ( n = 3). e-h Concentrations of IL-1β, IL-6, IL-8, and TNF-α were measured by ELISA ( n = 3). CCK-8: Cell Counting Kit-8; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; ELISA: enzyme-linked immunosorbent assay; IL: interleukin; LPS: lipopolysaccharide; RT-PCR: reverse transcription polymerase chain reaction; TNF-α: tumor necrosis factor alpha. ns, no significance, * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was purchased from Sigma.

    Techniques: CCK-8 Assay, Flow Cytometry, Cytometry, Expressing, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Counting, Reverse Transcription Polymerase Chain Reaction

    Na v 1.1 levels are reduced in Scn1a RX/+ mice, and this reduction is not prevented by tau ablation. Levels of Na v 1.1 and total sodium channels (pan Na v ) in the parietal cortex of 8-month-old mice were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels were used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype). The average Na v 1.1 to pan Na v ratio in Scn1a +/+ / Tau +/+ mice was arbitrarily defined as 1.0. ***p

    Journal: Annals of Neurology

    Article Title: Tau Reduction Prevents Disease in a Mouse Model of Dravet Syndrome

    doi: 10.1002/ana.24230

    Figure Lengend Snippet: Na v 1.1 levels are reduced in Scn1a RX/+ mice, and this reduction is not prevented by tau ablation. Levels of Na v 1.1 and total sodium channels (pan Na v ) in the parietal cortex of 8-month-old mice were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) levels were used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype). The average Na v 1.1 to pan Na v ratio in Scn1a +/+ / Tau +/+ mice was arbitrarily defined as 1.0. ***p

    Article Snippet: After blocking for 1 hour in 5% bovine serum albumin diluted in Tris-buffered saline (BSA-TBS), membranes were incubated overnight at 4°C in anti-Nav 1.1 (1:1,000; Alomone Labs, Jerusalem, Israel), anti–pan-sodium channel (Pan Nav, 1:1,000; Sigma), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:10,000; Millipore, Billerica, MA), anti-tau clone Tau-5 (1:3,000; Life Technologies), anti-tau clone EP2456Y (1:1,000; Millipore), anti–phospho-tau Ser 396/404 clone PHF-1 (1:3,000, a gift from Dr P. Davies), anti–phospho-tau Thr231 clone CP9 (1:25, a gift from Dr P. Davies), or anti–phospho-PHF-tau pSer202+Thr205 clone AT8 (1:80; Thermo Scientific, Waltham, MA).

    Techniques: Mouse Assay, Western Blot

    Cortical levels of total and phosphorylated tau are not altered in Scn1a RX/+ mice. Levels of phospho-tau (PHF-1, Ser396/Ser404; AT8, Ser202/Thr205; CP9, Thr231) and total tau (Tau-5, EP2456Y) in the parietal cortex of 8-month-old mice of the indicated genotypes were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype) revealed no statistically significant differences between Scn1a RX/+ /Tau +/+ and Scn1a +/+ / Tau +/+ mice (Student t test). Average phospho-tau to EP2456Y ratios (PHF-1, AT8, CP9) or average total tau levels (Tau-5, EP2456Y) in Scn1a +/+ / Tau +/+ mice were arbitrarily defined as 1.0. Values represent mean ± standard error of the mean.

    Journal: Annals of Neurology

    Article Title: Tau Reduction Prevents Disease in a Mouse Model of Dravet Syndrome

    doi: 10.1002/ana.24230

    Figure Lengend Snippet: Cortical levels of total and phosphorylated tau are not altered in Scn1a RX/+ mice. Levels of phospho-tau (PHF-1, Ser396/Ser404; AT8, Ser202/Thr205; CP9, Thr231) and total tau (Tau-5, EP2456Y) in the parietal cortex of 8-month-old mice of the indicated genotypes were determined by Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. (A) Representative Western blot. (B) Quantification of Western blot signals (n = 5–7 mice per genotype) revealed no statistically significant differences between Scn1a RX/+ /Tau +/+ and Scn1a +/+ / Tau +/+ mice (Student t test). Average phospho-tau to EP2456Y ratios (PHF-1, AT8, CP9) or average total tau levels (Tau-5, EP2456Y) in Scn1a +/+ / Tau +/+ mice were arbitrarily defined as 1.0. Values represent mean ± standard error of the mean.

    Article Snippet: After blocking for 1 hour in 5% bovine serum albumin diluted in Tris-buffered saline (BSA-TBS), membranes were incubated overnight at 4°C in anti-Nav 1.1 (1:1,000; Alomone Labs, Jerusalem, Israel), anti–pan-sodium channel (Pan Nav, 1:1,000; Sigma), anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:10,000; Millipore, Billerica, MA), anti-tau clone Tau-5 (1:3,000; Life Technologies), anti-tau clone EP2456Y (1:1,000; Millipore), anti–phospho-tau Ser 396/404 clone PHF-1 (1:3,000, a gift from Dr P. Davies), anti–phospho-tau Thr231 clone CP9 (1:25, a gift from Dr P. Davies), or anti–phospho-PHF-tau pSer202+Thr205 clone AT8 (1:80; Thermo Scientific, Waltham, MA).

    Techniques: Mouse Assay, Western Blot

    Effect of aptamers on 3D pol expression in BHK-21 cells transfected with pGFP-PAC replicon. Cell extracts were prepared at 4 h post-transfection. (a) Aliquots were analysed by SDS-PAGE and immunoblotting for the presence of FMDV 3D pol and cellular glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Cells transfected with a Δ3D pol replicon (pGFP-PAC-Δ3D), mock-transfected or transfected with cRNA were included as controls. (b) Densitometry was conducted using ImageJ imaging analysis software on triplicate experiments for 3D pol expression, normalized to GAPDH. Data show mean values with sd ( n = 3) and statistical analysis performed using two-tailed paired t -test (* = P

    Journal: The Journal of General Virology

    Article Title: Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers

    doi: 10.1099/vir.0.067751-0

    Figure Lengend Snippet: Effect of aptamers on 3D pol expression in BHK-21 cells transfected with pGFP-PAC replicon. Cell extracts were prepared at 4 h post-transfection. (a) Aliquots were analysed by SDS-PAGE and immunoblotting for the presence of FMDV 3D pol and cellular glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Cells transfected with a Δ3D pol replicon (pGFP-PAC-Δ3D), mock-transfected or transfected with cRNA were included as controls. (b) Densitometry was conducted using ImageJ imaging analysis software on triplicate experiments for 3D pol expression, normalized to GAPDH. Data show mean values with sd ( n = 3) and statistical analysis performed using two-tailed paired t -test (* = P

    Article Snippet: Cellular glyceraldehyde 3-phosphate dehydrogenase (GAPDH), employed as an internal control protein, was detected with an mAb (GAPDH-71.1, Sigma-Aldrich).

    Techniques: Expressing, Transfection, SDS Page, Imaging, Software, Two Tailed Test

    Heat shock protein 70 (HSP70) expression and function manipulation affects Th17 gene expression. ( a ) Western blot analysis of HSP70 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) proteins in the EL4 TCR + cells that were transfected with HSP70 siRNA or control in relation to the parallel molecular weight (MW) marker separation blot. Representative results from two independent experiments are shown. Gene expression analysis of the EL4 TCR + cells following anti-CD3 stimulation that has been transfected with HSP70 siRNA ( b ) or treated with pifithrin-μ (PFTμ) (10 μM) or with a control (dimethyl sulfoxide; DMSO) ( c ). ( d ) HSP70 gene expression changes of the EL4 TCR + cells following anti-CD3 stimulation or/and PFTμ (10 μM) treatment. Data represent mean +/− SD. * p

    Journal: International Journal of Molecular Sciences

    Article Title: The Heat Shock Protein HSP70 Promotes Th17 Genes’ Expression via Specific Regulation of microRNA

    doi: 10.3390/ijms21082823

    Figure Lengend Snippet: Heat shock protein 70 (HSP70) expression and function manipulation affects Th17 gene expression. ( a ) Western blot analysis of HSP70 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) proteins in the EL4 TCR + cells that were transfected with HSP70 siRNA or control in relation to the parallel molecular weight (MW) marker separation blot. Representative results from two independent experiments are shown. Gene expression analysis of the EL4 TCR + cells following anti-CD3 stimulation that has been transfected with HSP70 siRNA ( b ) or treated with pifithrin-μ (PFTμ) (10 μM) or with a control (dimethyl sulfoxide; DMSO) ( c ). ( d ) HSP70 gene expression changes of the EL4 TCR + cells following anti-CD3 stimulation or/and PFTμ (10 μM) treatment. Data represent mean +/− SD. * p

    Article Snippet: Immunoblotting was performed with mouse primary antibodies to HSP70 or AGO2 (sc-24 and B-3 respectively, both from Santa Cruz Biotechnology, Dallas, TX, USA) and glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) (MAB374; Merck Millipore, Burlington, MA, USA), and visualized by G: BOX Chemi (Syngene, Cambridge, UK).

    Techniques: Expressing, Western Blot, Transfection, Molecular Weight, Marker

    Validation of sortilin antibodies using sortilin knockout (−/−) and wildtype (+/+) mouse brains, with a characterization of the normal expression pattern of sortilin in rodent cerebrum. Panel (A) is a schematic drawing of the human sortilin protein, with the extracellular, transmembrane and intracellular domains of varying lengths of amino acid (a.a.) residues as marked. The immunogenic synthetic peptide sequences for the extracellular and intracellular C-terminal domain antibodies are also provided. Panels (B,C) show representative immunoblot and immunolabeling results obtained with the goat antibody (Gt Ab) against the extracellular domain in sortilin +/+ and −/− brains. Panels (D,E) show the results obtained with the rabbit antibody (Rt Ab) against the intracellular C-terminal. Both antibodies label a ~100 kDa band in sortilin +/+, but not in −/−, lysates (B,D) . Several non-specific bands are visible in the immunoblot with the goat antibody, equally present in the sortilin +/+ and −/− lysates, by extending the time of film exposure (B) . With overexposure, two light bands at ~40 and ~15 kDa (pointed by arrow) are also noticeable in the immunoblot of sortilin +/+ lysates with the rabbit antibody (D) . Light microscopic images (C,E) show neuronal profiles in the subiculum (Sub) to CA1 transitional region labeled by both antibodies in sortilin +/+, but not in −/−, brain sections. Confocal immunofluorescent images show completely colocalized labeling by the two antibodies in cortical pyramidal-like neurons (F–I) , CA3 pyramidal neurons and granule cells of the dentate gyrus (DG) (J–M) in C57BL mouse brain, with granular elements seen intracellularly (I,M) . Western blot applications: 12% SDS-PAGE gel and 13 μg equal amount protein loading. Additional abbreviations: CC, corpus callosum; s.p., stratum pyramidale; GCL, granule cell layer; PC, Parietal cortex; I-III, cortical layers; GAPHD, glyceraldehyde-3-phosphate dehydrogenase. Scale bar = 200 μm in (C) applying to (E) ; 100 μm in (F) applying to (G,H,J–L) , equivalent to 25 μm for (I,M) .

    Journal: Frontiers in Neuroanatomy

    Article Title: Sortilin Fragments Deposit at Senile Plaques in Human Cerebrum

    doi: 10.3389/fnana.2017.00045

    Figure Lengend Snippet: Validation of sortilin antibodies using sortilin knockout (−/−) and wildtype (+/+) mouse brains, with a characterization of the normal expression pattern of sortilin in rodent cerebrum. Panel (A) is a schematic drawing of the human sortilin protein, with the extracellular, transmembrane and intracellular domains of varying lengths of amino acid (a.a.) residues as marked. The immunogenic synthetic peptide sequences for the extracellular and intracellular C-terminal domain antibodies are also provided. Panels (B,C) show representative immunoblot and immunolabeling results obtained with the goat antibody (Gt Ab) against the extracellular domain in sortilin +/+ and −/− brains. Panels (D,E) show the results obtained with the rabbit antibody (Rt Ab) against the intracellular C-terminal. Both antibodies label a ~100 kDa band in sortilin +/+, but not in −/−, lysates (B,D) . Several non-specific bands are visible in the immunoblot with the goat antibody, equally present in the sortilin +/+ and −/− lysates, by extending the time of film exposure (B) . With overexposure, two light bands at ~40 and ~15 kDa (pointed by arrow) are also noticeable in the immunoblot of sortilin +/+ lysates with the rabbit antibody (D) . Light microscopic images (C,E) show neuronal profiles in the subiculum (Sub) to CA1 transitional region labeled by both antibodies in sortilin +/+, but not in −/−, brain sections. Confocal immunofluorescent images show completely colocalized labeling by the two antibodies in cortical pyramidal-like neurons (F–I) , CA3 pyramidal neurons and granule cells of the dentate gyrus (DG) (J–M) in C57BL mouse brain, with granular elements seen intracellularly (I,M) . Western blot applications: 12% SDS-PAGE gel and 13 μg equal amount protein loading. Additional abbreviations: CC, corpus callosum; s.p., stratum pyramidale; GCL, granule cell layer; PC, Parietal cortex; I-III, cortical layers; GAPHD, glyceraldehyde-3-phosphate dehydrogenase. Scale bar = 200 μm in (C) applying to (E) ; 100 μm in (F) applying to (G,H,J–L) , equivalent to 25 μm for (I,M) .

    Article Snippet: Separated protein products were electrotransferred to Trans-Blot pure nitrocellulose membranes, which were immunoblotted with the aforementioned antibodies (rabbit and goat anti-sortilin diluted at 1:2000, 6E10 at 1:4000, BACE1 at 1:2000 and phosphorylated tau at 1:2000; Cai et al., ), and that for loading controls including β-tubulin-III (1:5000, Millipore), β-actin (1:5000, Millipore) or glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:5000, Millipore).

    Techniques: Knock-Out, Expressing, Immunolabeling, Labeling, Western Blot, SDS Page