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  • 86
    Thermo Fisher qt01658692 gapdh
    Qt01658692 Gapdh, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore gapdh
    Western blot of vascular markers <t>VEGFR2,</t> VE-cadherin, vWF, and DLL4 of SPEC and FOS during in vitro assembly. <t>GAPDH</t> house-keeping protein functioned as the loading control. Anti-Dll4 blotting corresponding to endothelial tip cell phenotype was tracked across day 1, 2, and 3 of SPEC and FOS growth. Relative optical density was calculated by normalizing to FOS day 3 controls. Ratio of relative optical density for DLL4 to the relative optical density of GAPDH is displayed ( n = 3). Dll4 peaks at D2 and decreases at D3 are similarly reflected by VE-cadherin western blot. This suggests a period of quiescence after 2 days incubation of the SPECs. VEGFR2 and vWF, markers of endothelial cells, remain stable after reaching a peak at D2, suggesting limited endothelial cell proliferation at D3. ns, no statistical significance.
    Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17016 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore g3p dehydrogenase
    A model illustrating glycerol-triggered modulation of root development. The diagram shows the different root patterns in the absence (left) or the presence (right) of glycerol. The middle section shows a condensed schematic of plant glycerol metabolism and three important genes in this study (red). Glycerol is phosphorylated to <t>G3P</t> by GLI1 and can also be generated by GPDHc1 via the reduction of DHAP. G3P is oxidized to DHAP by FAD-GPDH or dephosphorylate to glycerol by GPP. Exogenous glycerol treatment can cause modifications of multiple pathways, including increased G3P and reactive oxygen species (ROS) levels, reduced the phosphate level and expression of PIN1 and PIN7 . It also affected polar auxin transport and the root meristem activity, thus resulting in modified root growth and development. Abbreviations: GLI1, glycerol kinase; GPDHc1, cytosolic glycerol-3-phosphate dehydrogenase; FAD-GPDH, flavin adenine dinucleotide-dependent glycerol-3-phosphate dehydrogenase; GPP, glycerol-3-phosphatase; G3P, glycerol-3-phosphate; DHAP, dihydroxyacetone phosphate; ATP, adenosine triphosphate; FAD, flavin adenine dinucleotide; NADH, the reduced form of nicotinamide adenine dinucleotide. For the sake of clarity, some co-substrates and/or co-products have been omitted in some reactions.
    G3p Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher gapdh
    Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and <t>c-fos</t> (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene <t>GAPDH.</t> Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p
    Gapdh, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 14561 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    AstraZeneca g3p
    Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and <t>c-fos</t> (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene <t>GAPDH.</t> Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p
    G3p, supplied by AstraZeneca, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    BioVision g3p detection assay buffer
    Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and <t>c-fos</t> (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene <t>GAPDH.</t> Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p
    G3p Detection Assay Buffer, supplied by BioVision, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    American Radiolabeled Chemicals Inc g3p
    Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and <t>c-fos</t> (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene <t>GAPDH.</t> Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p
    G3p, supplied by American Radiolabeled Chemicals Inc, used in various techniques. Bioz Stars score: 89/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore anti gapdh antibody
    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) <t>FGF2/GAPDH</t> immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.
    Anti Gapdh Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6371 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    American Radiolabeled Chemicals Inc h g3p
    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) <t>FGF2/GAPDH</t> immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.
    H G3p, supplied by American Radiolabeled Chemicals Inc, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Western blot of vascular markers VEGFR2, VE-cadherin, vWF, and DLL4 of SPEC and FOS during in vitro assembly. GAPDH house-keeping protein functioned as the loading control. Anti-Dll4 blotting corresponding to endothelial tip cell phenotype was tracked across day 1, 2, and 3 of SPEC and FOS growth. Relative optical density was calculated by normalizing to FOS day 3 controls. Ratio of relative optical density for DLL4 to the relative optical density of GAPDH is displayed ( n = 3). Dll4 peaks at D2 and decreases at D3 are similarly reflected by VE-cadherin western blot. This suggests a period of quiescence after 2 days incubation of the SPECs. VEGFR2 and vWF, markers of endothelial cells, remain stable after reaching a peak at D2, suggesting limited endothelial cell proliferation at D3. ns, no statistical significance.

    Journal: BioResearch Open Access

    Article Title: Vascular Tissue Engineering Using Scaffold-Free Prevascular Endothelial–Fibroblast Constructs

    doi: 10.1089/biores.2018.0039

    Figure Lengend Snippet: Western blot of vascular markers VEGFR2, VE-cadherin, vWF, and DLL4 of SPEC and FOS during in vitro assembly. GAPDH house-keeping protein functioned as the loading control. Anti-Dll4 blotting corresponding to endothelial tip cell phenotype was tracked across day 1, 2, and 3 of SPEC and FOS growth. Relative optical density was calculated by normalizing to FOS day 3 controls. Ratio of relative optical density for DLL4 to the relative optical density of GAPDH is displayed ( n = 3). Dll4 peaks at D2 and decreases at D3 are similarly reflected by VE-cadherin western blot. This suggests a period of quiescence after 2 days incubation of the SPECs. VEGFR2 and vWF, markers of endothelial cells, remain stable after reaching a peak at D2, suggesting limited endothelial cell proliferation at D3. ns, no statistical significance.

    Article Snippet: Following protein separation and overnight transfer onto PVDF membranes, western blots were performed using antibodies toward GAPDH (loading control) (Calbiochem CB1001; 1:1000), VEGFR2 (Abcam; ab39256, 1:900), VE-Cadherin (ThermoFisher Scientific 36-1900; 1:250), vWF (Abcam; ab6994, 1:500), and DLL4 (Abcam; ab7280, 1:1000).

    Techniques: Western Blot, In Vitro, Incubation

    Re-examining the existence and function of ‘mitochondrial STAT3.’ (A) Purification of mitochondria by sonication. (B) Purification of mitochondria by trypsinization. (C) Purification of mitochondria by washing with high concentration of salt combined with trypsinization. GRP78 was used as the ER marker; ATP5A, NDUFA9, NDUFA13, and VDAC were used as the mitochondrial marker; GAPDH was used as the cytosolic marker. (D) Sucrose density centrifuge of digitonin-solubilized mitochondria followed by Western blot analysis of STAT3 and mitochondrial complexes protein. (E) Co-immunoprecipitation experiment in digitonin-solubilized crude mitochondria. (F) ChIP-qPCR detection of STAT3-binding on mitochondrial DNA in mouse embryonic stem cells. (G) Serum reintroduction experiment in Neuro2A cells. (H) Quantification of Western blot results (G) in three independent experiments. * p

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: STAT3 Localizes in Mitochondria-Associated ER Membranes Instead of in Mitochondria

    doi: 10.3389/fcell.2020.00274

    Figure Lengend Snippet: Re-examining the existence and function of ‘mitochondrial STAT3.’ (A) Purification of mitochondria by sonication. (B) Purification of mitochondria by trypsinization. (C) Purification of mitochondria by washing with high concentration of salt combined with trypsinization. GRP78 was used as the ER marker; ATP5A, NDUFA9, NDUFA13, and VDAC were used as the mitochondrial marker; GAPDH was used as the cytosolic marker. (D) Sucrose density centrifuge of digitonin-solubilized mitochondria followed by Western blot analysis of STAT3 and mitochondrial complexes protein. (E) Co-immunoprecipitation experiment in digitonin-solubilized crude mitochondria. (F) ChIP-qPCR detection of STAT3-binding on mitochondrial DNA in mouse embryonic stem cells. (G) Serum reintroduction experiment in Neuro2A cells. (H) Quantification of Western blot results (G) in three independent experiments. * p

    Article Snippet: Samples were then transferred to PVDF membrane (Thermo) and immunoblotted using anti-NDUFA9 (Invitrogen), anti-NDUFS3 (Invitrogen), anti-NDUFA13 (Invitrogen), anti-ATP5A (Invitrogen), anti-SDHA (CST), anti-VDAC (CST), anti-HSP60 (CST), anti-PHB1 (CST), anti-PDH (CST), anti-GAPDH (Sigma), anti-GRP78 (SCBT), anti-ABCA1 (SCBT), anti-FLOT1 (CST), anti-STAT3 (CST), anti-STAT1 (SCBT), anti-AMPK (CST), anti-ERK1/2 (CST), anti-p38 (CST), anti-SYP (Abcam), anti-ACSL4 (Abcam), all diluted in 5% BSA:TBST at 1:1000, followed by appropriate HRP-conjugated secondary antibodies (Thermo) incubation (diluted in 5% non-fat milk:TBST at 1:10,000), and developed using the SuperSignalTM West Femto Maximum Sensitivity Substrate (Thermo).

    Techniques: Purification, Sonication, Concentration Assay, Marker, Western Blot, Immunoprecipitation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay

    A model illustrating glycerol-triggered modulation of root development. The diagram shows the different root patterns in the absence (left) or the presence (right) of glycerol. The middle section shows a condensed schematic of plant glycerol metabolism and three important genes in this study (red). Glycerol is phosphorylated to G3P by GLI1 and can also be generated by GPDHc1 via the reduction of DHAP. G3P is oxidized to DHAP by FAD-GPDH or dephosphorylate to glycerol by GPP. Exogenous glycerol treatment can cause modifications of multiple pathways, including increased G3P and reactive oxygen species (ROS) levels, reduced the phosphate level and expression of PIN1 and PIN7 . It also affected polar auxin transport and the root meristem activity, thus resulting in modified root growth and development. Abbreviations: GLI1, glycerol kinase; GPDHc1, cytosolic glycerol-3-phosphate dehydrogenase; FAD-GPDH, flavin adenine dinucleotide-dependent glycerol-3-phosphate dehydrogenase; GPP, glycerol-3-phosphatase; G3P, glycerol-3-phosphate; DHAP, dihydroxyacetone phosphate; ATP, adenosine triphosphate; FAD, flavin adenine dinucleotide; NADH, the reduced form of nicotinamide adenine dinucleotide. For the sake of clarity, some co-substrates and/or co-products have been omitted in some reactions.

    Journal: PLoS ONE

    Article Title: Glycerol Affects Root Development through Regulation of Multiple Pathways in Arabidopsis

    doi: 10.1371/journal.pone.0086269

    Figure Lengend Snippet: A model illustrating glycerol-triggered modulation of root development. The diagram shows the different root patterns in the absence (left) or the presence (right) of glycerol. The middle section shows a condensed schematic of plant glycerol metabolism and three important genes in this study (red). Glycerol is phosphorylated to G3P by GLI1 and can also be generated by GPDHc1 via the reduction of DHAP. G3P is oxidized to DHAP by FAD-GPDH or dephosphorylate to glycerol by GPP. Exogenous glycerol treatment can cause modifications of multiple pathways, including increased G3P and reactive oxygen species (ROS) levels, reduced the phosphate level and expression of PIN1 and PIN7 . It also affected polar auxin transport and the root meristem activity, thus resulting in modified root growth and development. Abbreviations: GLI1, glycerol kinase; GPDHc1, cytosolic glycerol-3-phosphate dehydrogenase; FAD-GPDH, flavin adenine dinucleotide-dependent glycerol-3-phosphate dehydrogenase; GPP, glycerol-3-phosphatase; G3P, glycerol-3-phosphate; DHAP, dihydroxyacetone phosphate; ATP, adenosine triphosphate; FAD, flavin adenine dinucleotide; NADH, the reduced form of nicotinamide adenine dinucleotide. For the sake of clarity, some co-substrates and/or co-products have been omitted in some reactions.

    Article Snippet: The DHAP content was assayed in reaction buffer (50 mM HEPES-NaOH, 1 mM MgCl2 and 10 µM NADH) containing 0.75 units of G3P dehydrogenase (Sigma) .

    Techniques: Generated, Expressing, Activity Assay, Modification

    G3P levels in seedlings treated with glycerol (nmol g −1 FW). (A) Wild-type seedlings were grown on agar plates containing 0.5×Murashige and Skoog (MS) medium plus 1% (w/v) sucrose in the absence or presence of 1 mM glycerol from 1–5 days post-germination (dpg). The G3P levels of the seedlings were examined. The data are presented as the mean ± SE (n = 3–4). The asterisks indicate significant differences between the means as determined by Student’s t-test (control versus 1 mM glycerol: *, p

    Journal: PLoS ONE

    Article Title: Glycerol Affects Root Development through Regulation of Multiple Pathways in Arabidopsis

    doi: 10.1371/journal.pone.0086269

    Figure Lengend Snippet: G3P levels in seedlings treated with glycerol (nmol g −1 FW). (A) Wild-type seedlings were grown on agar plates containing 0.5×Murashige and Skoog (MS) medium plus 1% (w/v) sucrose in the absence or presence of 1 mM glycerol from 1–5 days post-germination (dpg). The G3P levels of the seedlings were examined. The data are presented as the mean ± SE (n = 3–4). The asterisks indicate significant differences between the means as determined by Student’s t-test (control versus 1 mM glycerol: *, p

    Article Snippet: The DHAP content was assayed in reaction buffer (50 mM HEPES-NaOH, 1 mM MgCl2 and 10 µM NADH) containing 0.75 units of G3P dehydrogenase (Sigma) .

    Techniques: Mass Spectrometry

    Measurement of G3P binding to GlpT at various temperatures using intrinsic tryptophan fluorescence quenching. G3P was titrated until fluorescence quenching was saturated (about 15 μ M). The dissociation constant of G3P binding was found to be about 0.7 μ M, and substrate binding did not show temperature dependence. Data points represent the mean ± SE of three separate measurements.

    Journal: Biochemistry

    Article Title: Kinetic Evidence Is Consistent with the Rocker-Switch Mechanism of Membrane Transport by GlpT †

    doi: 10.1021/bi701383g

    Figure Lengend Snippet: Measurement of G3P binding to GlpT at various temperatures using intrinsic tryptophan fluorescence quenching. G3P was titrated until fluorescence quenching was saturated (about 15 μ M). The dissociation constant of G3P binding was found to be about 0.7 μ M, and substrate binding did not show temperature dependence. Data points represent the mean ± SE of three separate measurements.

    Article Snippet: For kinetics experiments, after equilibration of the whole cells to the desired temperature, substrate transport was initiated by the addition of [14 C]G3P (Sigma, St. Louis, MO), ranging from 6.25 to 400 μ M in a final reaction volume of 110 μ L (104.5 μ L of cells and 5.5 μ L of [14 C]G3P).

    Techniques: Binding Assay, Fluorescence

    Crystal structure of GlpT and the rocker-switch mechanism. (A) Ribbon diagram of GlpT viewed parallel to the membrane. The molecule consists of 12 transmembrane α-helices. The N-terminal domain is colored green, and the C-terminal domain is colored pink. (B) Schematic diagram of the single-binding site, alternating the access mechanism with a rocker-switch type of movement for the GlpT-mediated G3P–P i exchange reaction. The diagram describes the proposed conformational changes that the transporter undergoes during the reaction cycle. C o represents the protein in the outward-facing conformation, and C i represents the inward-facing one. The G3P substrate is represented by a small disk and triangle, and P i is represented by a small disk. (C) Schematic free-energy diagram illustrating the energy levels of the different conformations of GlpT that occur during the transport reaction cycle under physiological conditions. In the absence of substrate binding, the energy barrier (represented by a dotted line) prevents the conformational interconversion between the C o and C i states of the transporter. Substrate binding lowers the energy barrier sufficiently to allow Brownian motion (kT) to drive the conformational interconversion. S denotes the substrate.

    Journal: Biochemistry

    Article Title: Kinetic Evidence Is Consistent with the Rocker-Switch Mechanism of Membrane Transport by GlpT †

    doi: 10.1021/bi701383g

    Figure Lengend Snippet: Crystal structure of GlpT and the rocker-switch mechanism. (A) Ribbon diagram of GlpT viewed parallel to the membrane. The molecule consists of 12 transmembrane α-helices. The N-terminal domain is colored green, and the C-terminal domain is colored pink. (B) Schematic diagram of the single-binding site, alternating the access mechanism with a rocker-switch type of movement for the GlpT-mediated G3P–P i exchange reaction. The diagram describes the proposed conformational changes that the transporter undergoes during the reaction cycle. C o represents the protein in the outward-facing conformation, and C i represents the inward-facing one. The G3P substrate is represented by a small disk and triangle, and P i is represented by a small disk. (C) Schematic free-energy diagram illustrating the energy levels of the different conformations of GlpT that occur during the transport reaction cycle under physiological conditions. In the absence of substrate binding, the energy barrier (represented by a dotted line) prevents the conformational interconversion between the C o and C i states of the transporter. Substrate binding lowers the energy barrier sufficiently to allow Brownian motion (kT) to drive the conformational interconversion. S denotes the substrate.

    Article Snippet: For kinetics experiments, after equilibration of the whole cells to the desired temperature, substrate transport was initiated by the addition of [14 C]G3P (Sigma, St. Louis, MO), ranging from 6.25 to 400 μ M in a final reaction volume of 110 μ L (104.5 μ L of cells and 5.5 μ L of [14 C]G3P).

    Techniques: Binding Assay

    ( A ) MET transcript levels, as measured by qRT–PCR, after ectopic expression of miR-34c mimic in PC3. Mean of quadruplicate is shown. HPRT and PGK1 were used as endogenous controls. ( B ) Western blot on the protein levels of MET after ectopically expressing miR-34c in PC3, PNT2, and DU145 cells. Both the 170 kDa precursor and the 140 kDa mature form of Met are detected. GAPDH and α -actinin were used as loading controls.

    Journal: British Journal of Cancer

    Article Title: The tumour suppressor miR-34c targets MET in prostate cancer cells

    doi: 10.1038/bjc.2013.449

    Figure Lengend Snippet: ( A ) MET transcript levels, as measured by qRT–PCR, after ectopic expression of miR-34c mimic in PC3. Mean of quadruplicate is shown. HPRT and PGK1 were used as endogenous controls. ( B ) Western blot on the protein levels of MET after ectopically expressing miR-34c in PC3, PNT2, and DU145 cells. Both the 170 kDa precursor and the 140 kDa mature form of Met are detected. GAPDH and α -actinin were used as loading controls.

    Article Snippet: The membrane was incubated with antibodies directed against MET (sc-10, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), GAPDH 1:20000 (a-GAPDH, MAB374, Chemicon, CA, USA), and α -actinin (sc-17829, Santa Cruz Biotechnology).

    Techniques: Quantitative RT-PCR, Expressing, Western Blot

    rAAV2/1-mediated delivery of miR-Atx3-148 to the cerebellum of SCA3/MJD84.2 mice leads to gene silencing of human mutant Ataxin-3 . ( a ) The U6-miRNA expression cassette was cloned into a recombinant AAV expression vector upstream of a CMV-driven hrGFP expression cassette. rAAV2/1 virus was delivered into the deep cerebellar nuclei (DCN) of SCA3/MJD84.2 transgenic mice. Purkinje neurons in the cerebellar cortex, which project their axons to the DCN, are also transduced by DCN-delivered rAAV2/1 via retrograde transport. ( b – e ) Cerebellar coronal sections were obtained from injected mice 8 weeks post-surgery. Confocal microscopy analysis revealed strong hrGFP expression throughout neurons in the DCN ( b , c ) and in cerebellar Purkinje neurons ( d , e ).Scale bar in ( b ) and ( d ) = 200 µm. Scale bar in ( c ) and ( e ) = 50 µm. ( f ) Mutant human ATXN3 mRNA levels were suppressed following delivery of the rAAV2/1 miR-Atx3-148 virus. In contrast, the steady state levels of mouse Atxn3 and Calbindin-1 mRNAs remained unchanged in the presence of rAAV-miR-Atx3-148 virus. ( g ) Cerebellar mutant Ataxin-3 protein expression was confirmed by western blot using the 1H9 antibody. This antibody recognizes human mutant Ataxin-3 (top band) and endogenous mouse ataxin-3 (lower band). Four control-injected (lanes 1–4) and four rAAV-miR-148-injected (lanes 5–8) SCA3/84.2 mice were compared. As a group, SCA3/84.2 mice injected with rAAV-miR-148 showed a significant reduction (~34%) in mutant Ataxin-3 levels when compared to control injected mice. Gapdh levels were used to normalize for protein loading.* P

    Journal: Molecular Therapy

    Article Title: Silencing Mutant ATXN3 Expression Resolves Molecular Phenotypes in SCA3 Transgenic Mice

    doi: 10.1038/mt.2013.152

    Figure Lengend Snippet: rAAV2/1-mediated delivery of miR-Atx3-148 to the cerebellum of SCA3/MJD84.2 mice leads to gene silencing of human mutant Ataxin-3 . ( a ) The U6-miRNA expression cassette was cloned into a recombinant AAV expression vector upstream of a CMV-driven hrGFP expression cassette. rAAV2/1 virus was delivered into the deep cerebellar nuclei (DCN) of SCA3/MJD84.2 transgenic mice. Purkinje neurons in the cerebellar cortex, which project their axons to the DCN, are also transduced by DCN-delivered rAAV2/1 via retrograde transport. ( b – e ) Cerebellar coronal sections were obtained from injected mice 8 weeks post-surgery. Confocal microscopy analysis revealed strong hrGFP expression throughout neurons in the DCN ( b , c ) and in cerebellar Purkinje neurons ( d , e ).Scale bar in ( b ) and ( d ) = 200 µm. Scale bar in ( c ) and ( e ) = 50 µm. ( f ) Mutant human ATXN3 mRNA levels were suppressed following delivery of the rAAV2/1 miR-Atx3-148 virus. In contrast, the steady state levels of mouse Atxn3 and Calbindin-1 mRNAs remained unchanged in the presence of rAAV-miR-Atx3-148 virus. ( g ) Cerebellar mutant Ataxin-3 protein expression was confirmed by western blot using the 1H9 antibody. This antibody recognizes human mutant Ataxin-3 (top band) and endogenous mouse ataxin-3 (lower band). Four control-injected (lanes 1–4) and four rAAV-miR-148-injected (lanes 5–8) SCA3/84.2 mice were compared. As a group, SCA3/84.2 mice injected with rAAV-miR-148 showed a significant reduction (~34%) in mutant Ataxin-3 levels when compared to control injected mice. Gapdh levels were used to normalize for protein loading.* P

    Article Snippet: Ataxin-3 levels were detected by western blotting using the 1H9 antibody as mentioned above and normalized for Gapdh levels assessed using a monoclonal anti-GAPDH antibody (1:3,000 dilution; Millipore).

    Techniques: Mouse Assay, Mutagenesis, Expressing, Clone Assay, Recombinant, Plasmid Preparation, Transgenic Assay, Injection, Confocal Microscopy, Western Blot

    Lab-on-a-disc scWestern analysis of 14 single cells from a sparse, 24-cell biospecimens. (A) A box-and-whisker plot of cell-size distribution of U251-GFP cells settled in microwells. (B) A false-color overlay of fluorescence micrographs from GFP (green), β-TUB (blue), GADPH (red), STAT3 (black) proteins with fluorescence intensity profile plots. (C) Box-and-whisker plots of STAT3, GFP, β-TUB, and GAPDH distributions are obtained from an area-under-the-curve analysis. Box ends represent 25th and 75th percentiles; IQR is the difference between the 25th and 75th percentiles; median value is the red line at box middle; whiskers spread to 95% confidence limits; and red dots indicate outliers.

    Journal: Lab on a chip

    Article Title: High-selectivity cytology via lab-on-a-disc western blotting of individual cells

    doi: 10.1039/c6lc01333c

    Figure Lengend Snippet: Lab-on-a-disc scWestern analysis of 14 single cells from a sparse, 24-cell biospecimens. (A) A box-and-whisker plot of cell-size distribution of U251-GFP cells settled in microwells. (B) A false-color overlay of fluorescence micrographs from GFP (green), β-TUB (blue), GADPH (red), STAT3 (black) proteins with fluorescence intensity profile plots. (C) Box-and-whisker plots of STAT3, GFP, β-TUB, and GAPDH distributions are obtained from an area-under-the-curve analysis. Box ends represent 25th and 75th percentiles; IQR is the difference between the 25th and 75th percentiles; median value is the red line at box middle; whiskers spread to 95% confidence limits; and red dots indicate outliers.

    Article Snippet: Primary GAPDH, β-Tubulin, turboGFP, STAT3, and H3K79me2 antibodies were purchased from Sigma (SAB2500450), Abcam (ab6046), Pierce (PA5–22688), Cell Signalling (9139), and Abcam (ab3594), respectively.

    Techniques: Whisker Assay, Fluorescence

    Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and c-fos (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene GAPDH. Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p

    Journal: Journal of Cardiothoracic Surgery

    Article Title: Multikinase inhibitor sorafenib prevents pressure overload-induced left ventricular hypertrophy in rats by blocking the c-Raf/ERK1/2 signaling pathway

    doi: 10.1186/1749-8090-9-81

    Figure Lengend Snippet: Sorafenib regulates the increased expression of nuclear transcription factors resulting from AB. mRNA of c-myc (A) and c-fos (B) and GATA4 (C) are measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR), and normalized to the reference gene GAPDH. Increased expression of pro-proliferative transcription factors c-myc (A) and c-fos (B) and pro-hypertrophic factor GATA4 (C) in AB as compared to sham-operated animals (p

    Article Snippet: Predeveloped primer probes were used for ANP (Rn00561661_m1), GATA4 (Rn01530459_m1), c-myc (Rn00561507_m1) and c-fos (Rn02396759_m1) as well as the reference genes 18S rRNA and GAPDH (all from Applied Biosystems).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Seven active chalcones up-regulate the GCLC and HO-1 mRNA levels in PC12 cells. Cells were pretreated with 10 µM chalcones or vehicle control for 6 h. The mRNA levels of antioxidant genes GCLC ( A ) and HO-1 ( B ) were measured by qRT-PCR. The mRNA values for each gene were normalized to those of internal control GAPDH (glyceraldehyde phosphate dehydrogenase) mRNA, and expressed as a ratio to DMSO. Each bar represents the mean ± SD of three separate experiments. Statistical significance relative to DMSO group is indicated as follows: * p

    Journal: International Journal of Molecular Sciences

    Article Title: Synthetic Chalcones with Potent Antioxidant Ability on H2O2-Induced Apoptosis in PC12 Cells

    doi: 10.3390/ijms151018525

    Figure Lengend Snippet: Seven active chalcones up-regulate the GCLC and HO-1 mRNA levels in PC12 cells. Cells were pretreated with 10 µM chalcones or vehicle control for 6 h. The mRNA levels of antioxidant genes GCLC ( A ) and HO-1 ( B ) were measured by qRT-PCR. The mRNA values for each gene were normalized to those of internal control GAPDH (glyceraldehyde phosphate dehydrogenase) mRNA, and expressed as a ratio to DMSO. Each bar represents the mean ± SD of three separate experiments. Statistical significance relative to DMSO group is indicated as follows: * p

    Article Snippet: Reverse transcription and amplification of target genes GCLC and HO-1 , as well as the internal control GAPDH , were performed according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA).

    Techniques: Quantitative RT-PCR

    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques:

    ( A ) Mean (±SEM) percent CS-elicited freezing in Experiment 2. Rats were in one of three groups: High fear ( n = 8), Low fear ( n = 9), CS only ( n = 5). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 2. Data are presented as a percentage of mean data from group CS only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 2. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent CS-elicited freezing in Experiment 2. Rats were in one of three groups: High fear ( n = 8), Low fear ( n = 9), CS only ( n = 5). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 2. Data are presented as a percentage of mean data from group CS only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 2. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques:

    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 3. Rats were in one of three groups: High fear ( n = 13), Low fear ( n = 13), MK801 ( n = 12). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 3. Data are presented as a percentage of mean data from group MK801. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 3. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 3. Rats were in one of three groups: High fear ( n = 13), Low fear ( n = 13), MK801 ( n = 12). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 3. Data are presented as a percentage of mean data from group MK801. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 3. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques: