g3p dehydrogenase  (Millipore)


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    Name:
    GAPDH
    Description:
    GAPDH Glyceraldehyde 3 phosphate dehydrogenase catalyzes the conversion of glyceraldehyde 3 phosphate into D glycerate 1 3 bisphosphate as part of the glycolysis pathway GAPDH has also been found to function in additional cellular process such as transcription apoptosis oxidative stress and ER to Golgi transport
    Catalog Number:
    g5262
    Price:
    None
    Applications:
    GAPDH protein is suitable for use as a molecular weight marker and protein standard for molecular biology applications, including western blotting and mass spectometry.
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    Structured Review

    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.
    GAPDH Glyceraldehyde 3 phosphate dehydrogenase catalyzes the conversion of glyceraldehyde 3 phosphate into D glycerate 1 3 bisphosphate as part of the glycolysis pathway GAPDH has also been found to function in additional cellular process such as transcription apoptosis oxidative stress and ER to Golgi transport
    https://www.bioz.com/result/g3p dehydrogenase/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    g3p dehydrogenase - by Bioz Stars, 2020-09
    95/100 stars

    Images

    1) Product Images from "Glycerol Affects Root Development through Regulation of Multiple Pathways in Arabidopsis"

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

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0086269

    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.
    Figure Legend 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.

    Techniques Used: 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
    Figure Legend 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

    Techniques Used: Mass Spectrometry

    Related Articles

    Co-Immunoprecipitation Assay:

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB
    Article Snippet: .. Antibodies The primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit). .. Proximity ligation assay (PLA) To create our PLA probes a rat anti-D1R antibody (Sigma, D2944) was conjugated with a PLUS oligonucleotide (Duolink® In Situ Probemaker PLUS DUO92009, Sigma-Olink) and a rabbit anti-D2R (Millipore, AB5084P) antibody with a MINUS oligonucleotide (Duolink® In Situ Probemaker MINUS DUO92010, Sigma-Olink) following manufacturer's instructions.

    Proximity Ligation Assay:

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB
    Article Snippet: .. Antibodies The primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit). .. Proximity ligation assay (PLA) To create our PLA probes a rat anti-D1R antibody (Sigma, D2944) was conjugated with a PLUS oligonucleotide (Duolink® In Situ Probemaker PLUS DUO92009, Sigma-Olink) and a rabbit anti-D2R (Millipore, AB5084P) antibody with a MINUS oligonucleotide (Duolink® In Situ Probemaker MINUS DUO92010, Sigma-Olink) following manufacturer's instructions.

    Immunohistochemistry:

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB
    Article Snippet: .. Antibodies The primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit). .. Proximity ligation assay (PLA) To create our PLA probes a rat anti-D1R antibody (Sigma, D2944) was conjugated with a PLUS oligonucleotide (Duolink® In Situ Probemaker PLUS DUO92009, Sigma-Olink) and a rabbit anti-D2R (Millipore, AB5084P) antibody with a MINUS oligonucleotide (Duolink® In Situ Probemaker MINUS DUO92010, Sigma-Olink) following manufacturer's instructions.

    Immunodetection:

    Article Title: Lactate and Choline Metabolites Detected In Vitro by Nuclear Magnetic Resonance Spectroscopy Are Potential Metabolic Biomarkers for PI3K Inhibition in Pediatric Glioblastoma
    Article Snippet: .. Immunodetection was performed using antibodies against pAKT (Ser473), total AKT, pRPS6 (Ser240/244), total RPS6, HK2 (Cell Signaling), CHKA (Sigma), GLUT1, LDHA (Santa Cruz Biotechnology) and GAPDH (Chemicon). .. Blots were revealed with peroxidase-conjugated secondary anti-rabbit or anti-mouse antibodies (Cell Signaling) followed by ECL chemiluminescence solution (Amersham Biosciences).

    Incubation:

    Article Title: Nuclear relocalisation of cytoplasmic poly(A)-binding proteins PABP1 and PABP4 in response to UV irradiation reveals mRNA-dependent export of metazoan PABPs
    Article Snippet: .. Membranes were subject to western blotting with primary antibodies, PABP1 (1:10,000), PABP4 (1:5000), GAPDH (1:1000), PARP (1:1000), α-tubulin (1:10,000), G3BP (1:2000) or RNAPII CTD pS5 (1:2000), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit (Sigma #A0545-1; 1:100,000) or goat anti-mouse (Pierce #31444; 1:20,000) secondary a ntibodies and developed using an enhanced chemiluminescence reagent (Amersham). .. [35 S]methionine labelling Cells cultured in six-well plates were labelled for 15 minutes with 50 μCi/ml [35 S]methionine (MP Biomedicals) in methionine-free minimal essential medium (MP Biomedicals).

    Western Blot:

    Article Title: Nuclear relocalisation of cytoplasmic poly(A)-binding proteins PABP1 and PABP4 in response to UV irradiation reveals mRNA-dependent export of metazoan PABPs
    Article Snippet: .. Membranes were subject to western blotting with primary antibodies, PABP1 (1:10,000), PABP4 (1:5000), GAPDH (1:1000), PARP (1:1000), α-tubulin (1:10,000), G3BP (1:2000) or RNAPII CTD pS5 (1:2000), followed by incubation with horseradish peroxidase-conjugated goat anti-rabbit (Sigma #A0545-1; 1:100,000) or goat anti-mouse (Pierce #31444; 1:20,000) secondary a ntibodies and developed using an enhanced chemiluminescence reagent (Amersham). .. [35 S]methionine labelling Cells cultured in six-well plates were labelled for 15 minutes with 50 μCi/ml [35 S]methionine (MP Biomedicals) in methionine-free minimal essential medium (MP Biomedicals).

    Article Title: Intracellular fatty acids suppress ?-adrenergic induction of PKA-targeted gene expression in white adipocytes
    Article Snippet: .. Western blot was performed using antibodies against ATGL , HSL (provided by Dr. A. Chaudhry) , and GAPDH (Millipore) as described ( ). .. Glycerol and fatty acids were quantified as stated above for animal studies. cAMP response element (CRE) reporter assays were performed using a cAMP-responsive reporter plasmid (pADneo2 C6-BGL) provided by Dr. Adolf Himmler ( ).

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  • 99
    Millipore anti gapdh
    Neuronal specificity and efficacy of RNA interference with <t>SUMO2/3</t> and its effects on survival of mouse primary cortical neurons. At days in vitro (DIV) 3, primary cortical neurons were transduced with lentiviral particles expressing EGFP as a reporter and either SUMO2/3 or control microRNA driven by the neuron-specific synapsin promoter. ( A ) Verification of transduction efficacy and neuronal specificity of cultures transduced with lentiviral particles driven by the synapsin promoter. Immunohistochemistry was performed after paraformaldehyde (PFA) fixation on DIV 12 with antibodies against EGFP, microtubuli-associated protein 2 (MAP2), and nuclear counterstain with DAPI. The multiplicity of infection (MOI) was ∼50. Scale bar=100 μ m. ( B ) Verification of knockdown efficiency of SUMO2/3 versus control microRNAs (1=LacZ, 2=non-targeting scrambled) with and without OGD (45 minutes and 3 hours reoxygenation) shown by a representative western blot analysis. Neuronal cultures were analyzed on DIV 12 and subjected to SDS-PAGE. Membranes were probed with antibodies against SUMO2/3, EGFP, and <t>GAPDH.</t> EGFP expression corresponded to an equal MOI of lentiviral particles and concomitant microRNA expression. GAPDH served as a housekeeping protein and equal loading control. ( C ) SUMO2/3 microRNA does not influence baseline survival over time up to DIV 12. In brief, microscopic pictures of EGFP fluorescence (indicative for microRNA delivery) were taken at DIV 6, 9, and 12 as described in the ‘Materials and methods' section. EGFP-expressing neurons were counted and ratios were calculated for DIV 9/6 (indicated in blue) and DIV 12/9 (red) to evaluate cell survival over time. We assumed an effect size > 0.15 and performed a prospective power analysis with α =0.05 and β =0.20. There was no significant difference between groups in a two-way repeated-measures ANOVA followed by Tukey's post hoc analysis. ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; EGFP, enhanced green fluorescent protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MAP2, microtubule-associated protein 2; OGD, oxygen–glucose deprivation; SUMO2/3, small ubiquitin-like modifier-2/3.
    Anti Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1300 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh/product/Millipore
    Average 99 stars, based on 1300 article reviews
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    anti gapdh - by Bioz Stars, 2020-09
    99/100 stars
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    99
    Millipore anti glyceraldehyde 3 phosphate dehydrogenase
    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. <t>Glyceraldehyde-3-phosphate</t> 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
    Anti Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti glyceraldehyde 3 phosphate dehydrogenase/product/Millipore
    Average 99 stars, based on 352 article reviews
    Price from $9.99 to $1999.99
    anti glyceraldehyde 3 phosphate dehydrogenase - by Bioz Stars, 2020-09
    99/100 stars
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    Neuronal specificity and efficacy of RNA interference with SUMO2/3 and its effects on survival of mouse primary cortical neurons. At days in vitro (DIV) 3, primary cortical neurons were transduced with lentiviral particles expressing EGFP as a reporter and either SUMO2/3 or control microRNA driven by the neuron-specific synapsin promoter. ( A ) Verification of transduction efficacy and neuronal specificity of cultures transduced with lentiviral particles driven by the synapsin promoter. Immunohistochemistry was performed after paraformaldehyde (PFA) fixation on DIV 12 with antibodies against EGFP, microtubuli-associated protein 2 (MAP2), and nuclear counterstain with DAPI. The multiplicity of infection (MOI) was ∼50. Scale bar=100 μ m. ( B ) Verification of knockdown efficiency of SUMO2/3 versus control microRNAs (1=LacZ, 2=non-targeting scrambled) with and without OGD (45 minutes and 3 hours reoxygenation) shown by a representative western blot analysis. Neuronal cultures were analyzed on DIV 12 and subjected to SDS-PAGE. Membranes were probed with antibodies against SUMO2/3, EGFP, and GAPDH. EGFP expression corresponded to an equal MOI of lentiviral particles and concomitant microRNA expression. GAPDH served as a housekeeping protein and equal loading control. ( C ) SUMO2/3 microRNA does not influence baseline survival over time up to DIV 12. In brief, microscopic pictures of EGFP fluorescence (indicative for microRNA delivery) were taken at DIV 6, 9, and 12 as described in the ‘Materials and methods' section. EGFP-expressing neurons were counted and ratios were calculated for DIV 9/6 (indicated in blue) and DIV 12/9 (red) to evaluate cell survival over time. We assumed an effect size > 0.15 and performed a prospective power analysis with α =0.05 and β =0.20. There was no significant difference between groups in a two-way repeated-measures ANOVA followed by Tukey's post hoc analysis. ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; EGFP, enhanced green fluorescent protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MAP2, microtubule-associated protein 2; OGD, oxygen–glucose deprivation; SUMO2/3, small ubiquitin-like modifier-2/3.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: SUMO2/3 conjugation is an endogenous neuroprotective mechanism

    doi: 10.1038/jcbfm.2011.112

    Figure Lengend Snippet: Neuronal specificity and efficacy of RNA interference with SUMO2/3 and its effects on survival of mouse primary cortical neurons. At days in vitro (DIV) 3, primary cortical neurons were transduced with lentiviral particles expressing EGFP as a reporter and either SUMO2/3 or control microRNA driven by the neuron-specific synapsin promoter. ( A ) Verification of transduction efficacy and neuronal specificity of cultures transduced with lentiviral particles driven by the synapsin promoter. Immunohistochemistry was performed after paraformaldehyde (PFA) fixation on DIV 12 with antibodies against EGFP, microtubuli-associated protein 2 (MAP2), and nuclear counterstain with DAPI. The multiplicity of infection (MOI) was ∼50. Scale bar=100 μ m. ( B ) Verification of knockdown efficiency of SUMO2/3 versus control microRNAs (1=LacZ, 2=non-targeting scrambled) with and without OGD (45 minutes and 3 hours reoxygenation) shown by a representative western blot analysis. Neuronal cultures were analyzed on DIV 12 and subjected to SDS-PAGE. Membranes were probed with antibodies against SUMO2/3, EGFP, and GAPDH. EGFP expression corresponded to an equal MOI of lentiviral particles and concomitant microRNA expression. GAPDH served as a housekeeping protein and equal loading control. ( C ) SUMO2/3 microRNA does not influence baseline survival over time up to DIV 12. In brief, microscopic pictures of EGFP fluorescence (indicative for microRNA delivery) were taken at DIV 6, 9, and 12 as described in the ‘Materials and methods' section. EGFP-expressing neurons were counted and ratios were calculated for DIV 9/6 (indicated in blue) and DIV 12/9 (red) to evaluate cell survival over time. We assumed an effect size > 0.15 and performed a prospective power analysis with α =0.05 and β =0.20. There was no significant difference between groups in a two-way repeated-measures ANOVA followed by Tukey's post hoc analysis. ANOVA, analysis of variance; DAPI, 4′,6-diamidino-2-phenylindole; EGFP, enhanced green fluorescent protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MAP2, microtubule-associated protein 2; OGD, oxygen–glucose deprivation; SUMO2/3, small ubiquitin-like modifier-2/3.

    Article Snippet: Immunoblots Neurons were harvested as described previously ( ) and probed with a SUMO2/3-specific antibody (Invitrogen; 1:1,000), anti-GAPDH (anti-glyceraldehyde 3-phosphate dehydrogenase, Millipore; 1:75,000), and anti-EGFP (Santa Cruz Biotechnology, 1:1,000).

    Techniques: In Vitro, Transduction, Expressing, Immunohistochemistry, Infection, Western Blot, SDS Page, Fluorescence

    Inhibition of Nox2-activity reduces oxidative stress and Src kinase-mediated impaired autophagy ( a ) Nox2-specific ROS production was assessed using the Nox2 redox biosensor p47-roGFP redox biosensor Cat: catalase, PEG-Cat: pegylated catalse. ( b ) Measurement of intracellular glutathione redox potential with Grx1-roGFP2. ( c ) Analysis of Rac1 and ( d ) Src. ( e ) Immunoblot of precipitated p47 phox probed with an anti-phosphoserine or anti-p47 phox antibody. ( f ) Nox2-specific intracellular ROS production was measured using p47-roGFP redox biosensor. ( g ) Extracellular ROS production was assessed using Amplex-red dye. ( h ) Plasma membrane calcium influx was measured by analyzing the Fura-2 fluorescence quench rate upon addition of extracellular Mn 2+ . ( i ) Intracellular RNS generation was measured using DAF-FM. Bars represent average ±SEM from n=15 individual fibers for each condition in ( a , b , f , g , j and i ). Markers of autophagy were analyzed in isolated fibers (incubated with or without PP2) from FDBs. ( k ) Autophagosome formation was analyzed using fluorescence microscopy (scale bar=100 μm) and illustrated LC3 localization and autophagosome formation. ( l ) Confocal microscopy detected p62-LC3 localization in single fibers from FDBs (scale bar=140 μm and 50 μm for white box areas). All immunoblots were performed with isolated proteins from FDBs and probed with antibodies as indicated. GAPDH was detected as a loading control. Representative images are shown. Bars represent average ±SEM from n=3 independent biological experiments. Statistical differences between groups were determined using ANOVA with Tukey’s post-hoc test. *p

    Journal: Nature communications

    Article Title: Src-dependent impairment of autophagy by oxidative stress in a mouse model of Duchenne muscular dystrophy

    doi: 10.1038/ncomms5425

    Figure Lengend Snippet: Inhibition of Nox2-activity reduces oxidative stress and Src kinase-mediated impaired autophagy ( a ) Nox2-specific ROS production was assessed using the Nox2 redox biosensor p47-roGFP redox biosensor Cat: catalase, PEG-Cat: pegylated catalse. ( b ) Measurement of intracellular glutathione redox potential with Grx1-roGFP2. ( c ) Analysis of Rac1 and ( d ) Src. ( e ) Immunoblot of precipitated p47 phox probed with an anti-phosphoserine or anti-p47 phox antibody. ( f ) Nox2-specific intracellular ROS production was measured using p47-roGFP redox biosensor. ( g ) Extracellular ROS production was assessed using Amplex-red dye. ( h ) Plasma membrane calcium influx was measured by analyzing the Fura-2 fluorescence quench rate upon addition of extracellular Mn 2+ . ( i ) Intracellular RNS generation was measured using DAF-FM. Bars represent average ±SEM from n=15 individual fibers for each condition in ( a , b , f , g , j and i ). Markers of autophagy were analyzed in isolated fibers (incubated with or without PP2) from FDBs. ( k ) Autophagosome formation was analyzed using fluorescence microscopy (scale bar=100 μm) and illustrated LC3 localization and autophagosome formation. ( l ) Confocal microscopy detected p62-LC3 localization in single fibers from FDBs (scale bar=140 μm and 50 μm for white box areas). All immunoblots were performed with isolated proteins from FDBs and probed with antibodies as indicated. GAPDH was detected as a loading control. Representative images are shown. Bars represent average ±SEM from n=3 independent biological experiments. Statistical differences between groups were determined using ANOVA with Tukey’s post-hoc test. *p

    Article Snippet: Anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase), anti-active Rac1, anti-p47phox and anti-P-serine were purchased from Millipore.

    Techniques: Inhibition, Activity Assay, Fluorescence, Isolation, Incubation, Microscopy, Confocal Microscopy, Western Blot

    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

    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