glyceraldehyde 3 phosphate dehydrogenase gapdh Millipore Search Results


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  • 99
    Millipore glyceraldehyde 3 phosphate dehydrogenase gapdh
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA glyceraldehyde 3 phosphate dehydrogenase gapdh
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore human glyceraldehyde 3 phosphate dehydrogenase gapdh
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Human Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mouse glyceraldehyde 3 phosphate dehydrogenase
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Mouse Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore internal control glyceraldehyde 3 phosphate dehydrogenase gapdh
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Internal Control Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) <t>GAPDH</t> in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.
    Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore protein glyceraldehyde 3 phosphate dehydrogenase gapdh
    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 <t>GAPDH</t> (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
    Protein Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA glyceraldehyde 3 phosphate dehydrogenase
    Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related <t>glyceraldehyde-3-phosphate</t> dehydrogenase. The results are represented as means ± SEM ( n = 4). # P
    Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 94/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore glyceraldehyde 3 phosphate dehydrogenase gapdh expression
    Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related <t>glyceraldehyde-3-phosphate</t> dehydrogenase. The results are represented as means ± SEM ( n = 4). # P
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Expression, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc glyceraldehyde 3 phosphate dehydrogenase gapdh
    Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related <t>glyceraldehyde-3-phosphate</t> dehydrogenase. The results are represented as means ± SEM ( n = 4). # P
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1835 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rabbit muscle glyceraldehyde 3 phosphate dehydrogenase gapdh
    PIO interacts with and inhibits purified <t>GAPDH.</t> (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.
    Rabbit Muscle Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti acetyl glyceraldehyde 3 phosphate dehydrogenase gapdh
    PIO interacts with and inhibits purified <t>GAPDH.</t> (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.
    Anti Acetyl Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA glyceraldehyde 3 phosphate dehydrogenase gapdh abs16
    PIO interacts with and inhibits purified <t>GAPDH.</t> (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Abs16, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 95/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    PIO interacts with and inhibits purified <t>GAPDH.</t> (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.
    Monoclonal Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore cardiac glyceraldehyde 3 phosphate dehydrogenase gapdh
    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 <t>(GAPDH)</t> as an internal control.
    Cardiac Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti glyceraldehyde 3 phosphate dehydrogenase gapdh peroxidase
    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 <t>(GAPDH)</t> as an internal control.
    Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Peroxidase, supplied by Millipore, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mouse anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Expression of TMEM106B in different cell types. ( A ) Cell lysates from HEK293T, NSC-34 and BV-2 were loaded and blotted with anti-TMEM106B and anti-glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH)</t> antibodies. ( B ) TMEM106B forms homodimers. FLAG-tagged
    Mouse Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rabbit anti glyceraldehyde 3 phosphate dehydrogenase gapdh
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Rabbit Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore monoclonal anti glyceraldehyde 3 phosphate dehydrogenase gapdh 1 100 000 millipore
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Monoclonal Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh 1 100 000 Millipore, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore anti glyceraldehyde 3 phosphate dehydrogenase gapdh polyclonal antibodies
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Anti Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Polyclonal Antibodies, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore human polyclonal glyceraldehyde 3 phosphate dehydrogenase gapdh
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Human Polyclonal Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore recombinant glyceraldehyde 3 phosphate dehydrogenase gapdh
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Recombinant Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore primary mouse glyceraldehyde 3 phosphate dehydrogenase gapdh antibody
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Primary Mouse Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary mouse glyceraldehyde 3 phosphate dehydrogenase gapdh antibody/product/Millipore
    Average 86 stars, based on 7 article reviews
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    90
    Millipore mouse monoclonal glyceraldehyde 3 phosphate dehydrogenase gapdh 1 1000
    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase <t>(GAPDH).</t> ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p
    Mouse Monoclonal Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh 1 1000, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal glyceraldehyde 3 phosphate dehydrogenase gapdh 1 1000/product/Millipore
    Average 90 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    mouse monoclonal glyceraldehyde 3 phosphate dehydrogenase gapdh 1 1000 - by Bioz Stars, 2020-04
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    Image Search Results


    Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) GAPDH in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Identification and isolation of kidney-derived stem cells from transgenic rats with diphtheria toxin-induced kidney damage

    doi: 10.3892/etm.2016.3516

    Figure Lengend Snippet: Reverse transcription-polymerase chain reaction analysis. Expression profile of (A) Pax-2, (B) Oct-4 and (C) GAPDH in the isolated kidney-derived stem cells. The reactions were run in triplicate. PAX-2, paired box 2; Oct-4, octamer-binding transcription factor 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

    Article Snippet: PCR was performed on 1 µl cDNA using the following primers: Oct-4, forward 5′-CTGTAACCGGCGCCAGAA-3′ and reverse 5′-TGCATGGGAGAGCCCAGA-3′ (Sigma-Aldrich); Pax-2, the SABiosciences RT2 PCR primer set (LOC293992; Qiagen Sciences, LLC); and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), forward 5′-TGGAGAGGCCTGCCAAGTA-3′ and reverse 5′-AAGAGTGGGAGTTGCTGTTG-3′ (Sigma-Aldrich).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Isolation, Derivative Assay, Binding Assay

    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

    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

    non-structural protein 1 (NSP1) has a low expression level that increases late in infection in rhesus rotavirus (RRV)-infected cells. MA-104 cells were mock-infected or infected with RRV at a multiplicity of infection of 10 and the cells were processed using Western blot analysis with anti-NSP1, anti-NSP2 and anti- glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at different times post-infection (pi) (as indicated). The anti-NSP1 serum detected an intense cellular band in mock-infected cells (Lane 1, panel A), whereas a specific NSP1 band was detected only in RRV-infected cells (A). The results of one representative WB assay are shown (B), with two lengths of film exposure [low (NSP1 L-EXP) or high (NSP1 H-EXP)], as well as quantitative analyses of three independent experiments normalising the band intensities of NSP1 to the loading control GAPDH (C). Standard deviations are shown above each bar. hpi: hours pi.

    Journal: Memórias do Instituto Oswaldo Cruz

    Article Title: The shift from low to high non-structural protein 1 expression in rotavirus-infected MA-104 cells

    doi: 10.1590/0074-0276108042013005

    Figure Lengend Snippet: non-structural protein 1 (NSP1) has a low expression level that increases late in infection in rhesus rotavirus (RRV)-infected cells. MA-104 cells were mock-infected or infected with RRV at a multiplicity of infection of 10 and the cells were processed using Western blot analysis with anti-NSP1, anti-NSP2 and anti- glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at different times post-infection (pi) (as indicated). The anti-NSP1 serum detected an intense cellular band in mock-infected cells (Lane 1, panel A), whereas a specific NSP1 band was detected only in RRV-infected cells (A). The results of one representative WB assay are shown (B), with two lengths of film exposure [low (NSP1 L-EXP) or high (NSP1 H-EXP)], as well as quantitative analyses of three independent experiments normalising the band intensities of NSP1 to the loading control GAPDH (C). Standard deviations are shown above each bar. hpi: hours pi.

    Article Snippet: The membranes were blocked with 5% non-fat dry milk in Tris-buffered saline + 0.15% Tween 20 (TBS-Tween) at room temperature (RT) and further incubated for 2.5 h at RT with anti-NSP1 (1:500 or 1:5,000) or 1 h with anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (MAB374, Millipore, Billerica, MA) (1:10,000), both of which were diluted with 1% non-fat dry milk in TBS-Tween.

    Techniques: Expressing, Infection, Western Blot

    Western blotting of the mitochondrial respiratory complexes I–V in the cortex of control and curcumin-supplemented APOE3 mice. Relative intensities of bands were quantified by densitometry and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served

    Journal: Genes & Nutrition

    Article Title: Adenosine triphosphate concentrations are higher in the brain of APOE3- compared to APOE4-targeted replacement mice and can be modulated by curcumin

    doi: 10.1007/s12263-014-0397-3

    Figure Lengend Snippet: Western blotting of the mitochondrial respiratory complexes I–V in the cortex of control and curcumin-supplemented APOE3 mice. Relative intensities of bands were quantified by densitometry and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served

    Article Snippet: Proteins were then transferred to PVDF membranes (Amersham Biosciences) and incubated with the respective primary antibodies: MitoProfile Total OXPHOS (ab110411), complex IV (ab110259) (both from Abcam, Cambridge, UK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (MAB374) (Millipore, Billerica, USA), and secondary antibodies (Calbiochem, Germany) conjugated with horseradish peroxidase.

    Techniques: Western Blot, Mouse Assay

    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

    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

    IL-22 mRNA decay is accelerated by U0126. ( a – c ) Jurkat T cells were either kept as unstimulated control (Co) or stimulated with TPA (100 ng/ml)/A23187 (10 μM) for 4 h. ( a ) Thereafter, IL-22 mRNA expression was assessed. One representative of three independently performed experiments is shown in which IL-22 mRNA is determined by standard PCR and realtime PCR (normalized to GAPDH with fold-induction versus control), respectively. All cultures were adjusted to a final concentration of 0.11% DMSO (vehicle for TPA/A23187). ( b , c ) After the 4 h induction period, cells were washed twice with PBS. Then actinomycin (Act) D (0.5 μg/ml) and, where indicated, U0126 (10 μM) was added. All cultures were adjusted to a final concentration of 0.22% DMSO (vehicle for TPA/A23187, Act D, U0126). ( b ) IL-22 mRNA levels were determined by realtime PCR at the indicated time points. IL-22 mRNA was normalized to that of GAPDH. Data depicted (as % of IL-22 mRNA expression after TPA/A23187 at t 0 , the time point of Act D addition) are expressed as means ± SD (n = 3; *p = 0.0215). Statistical analysis on percent data, Student’s t-test. ( c ) After a total incubation time of 8 h (4 h induction period using TPA/A23187 followed by 4 h of incubation with Act D in presence or absence of U0126), cell viability was determined and is shown as percent of untreated control (n = 3).

    Journal: Scientific Reports

    Article Title: Tristetraprolin regulation of interleukin-22 production

    doi: 10.1038/srep15112

    Figure Lengend Snippet: IL-22 mRNA decay is accelerated by U0126. ( a – c ) Jurkat T cells were either kept as unstimulated control (Co) or stimulated with TPA (100 ng/ml)/A23187 (10 μM) for 4 h. ( a ) Thereafter, IL-22 mRNA expression was assessed. One representative of three independently performed experiments is shown in which IL-22 mRNA is determined by standard PCR and realtime PCR (normalized to GAPDH with fold-induction versus control), respectively. All cultures were adjusted to a final concentration of 0.11% DMSO (vehicle for TPA/A23187). ( b , c ) After the 4 h induction period, cells were washed twice with PBS. Then actinomycin (Act) D (0.5 μg/ml) and, where indicated, U0126 (10 μM) was added. All cultures were adjusted to a final concentration of 0.22% DMSO (vehicle for TPA/A23187, Act D, U0126). ( b ) IL-22 mRNA levels were determined by realtime PCR at the indicated time points. IL-22 mRNA was normalized to that of GAPDH. Data depicted (as % of IL-22 mRNA expression after TPA/A23187 at t 0 , the time point of Act D addition) are expressed as means ± SD (n = 3; *p = 0.0215). Statistical analysis on percent data, Student’s t-test. ( c ) After a total incubation time of 8 h (4 h induction period using TPA/A23187 followed by 4 h of incubation with Act D in presence or absence of U0126), cell viability was determined and is shown as percent of untreated control (n = 3).

    Article Snippet: Analysis of IL-22 and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mRNA by standard PCR Total RNA, isolated by Tri-Reagent (Sigma-Aldrich) was transcribed using random hexameric primers (Qiagen, Hilden, Germany) and Moloney virus reverse transcriptase (Life Techno-logies).

    Techniques: Expressing, Polymerase Chain Reaction, Concentration Assay, Activated Clotting Time Assay, Incubation

    Splenic CD3 + T cells from TTP −/− -mice display prolonged IL-22 mRNA half-life. ( a ) Splenic CD3 + T-cells were isolated from TTP −/− mice (n = 14) and wildtype (wt) littermates (n = 11). Cells of individual mice were either kept as unstimulated control (Co) or stimulated with αCD3 (15 μg/ml)/αCD28 (1.5 μg/ml). After 4 h, IL-22 mRNA was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH (means ± SEM; ***p

    Journal: Scientific Reports

    Article Title: Tristetraprolin regulation of interleukin-22 production

    doi: 10.1038/srep15112

    Figure Lengend Snippet: Splenic CD3 + T cells from TTP −/− -mice display prolonged IL-22 mRNA half-life. ( a ) Splenic CD3 + T-cells were isolated from TTP −/− mice (n = 14) and wildtype (wt) littermates (n = 11). Cells of individual mice were either kept as unstimulated control (Co) or stimulated with αCD3 (15 μg/ml)/αCD28 (1.5 μg/ml). After 4 h, IL-22 mRNA was determined by realtime PCR. IL-22 mRNA was normalized to that of GAPDH (means ± SEM; ***p

    Article Snippet: Analysis of IL-22 and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mRNA by standard PCR Total RNA, isolated by Tri-Reagent (Sigma-Aldrich) was transcribed using random hexameric primers (Qiagen, Hilden, Germany) and Moloney virus reverse transcriptase (Life Techno-logies).

    Techniques: Mouse Assay, Isolation, Polymerase Chain Reaction

    MAP kinase signaling is essential to IL-22 mRNA induction. ( a – c ) Where indicated, Jurkat T cells were pretreated with MAP kinase inhibitors for 1 h prior to stimulation with TPA (100 ng/ml)/A23187 (10 μM) for 4 h. PD98059 (n = 7), 50 μM; U0126 (n = 3), 10 μM, FR180204 (n = 11), 10 μM; SP600125 (n = 3), 10 μM; SB203580 (n = 5), 10 μM. In addition, cell were kept as unstimulated control. All cultures were adjusted to a final concentration of 0.21% DMSO (vehicle for TPA/A23187 plus inhibitor). ( a , c ) IL-22 and IL-6 mRNA were determined by realtime PCR. Target mRNA was normalized to GAPDH. Data are depicted as % of TPA/A23187-stimulation (means ± SD; * p

    Journal: Scientific Reports

    Article Title: Tristetraprolin regulation of interleukin-22 production

    doi: 10.1038/srep15112

    Figure Lengend Snippet: MAP kinase signaling is essential to IL-22 mRNA induction. ( a – c ) Where indicated, Jurkat T cells were pretreated with MAP kinase inhibitors for 1 h prior to stimulation with TPA (100 ng/ml)/A23187 (10 μM) for 4 h. PD98059 (n = 7), 50 μM; U0126 (n = 3), 10 μM, FR180204 (n = 11), 10 μM; SP600125 (n = 3), 10 μM; SB203580 (n = 5), 10 μM. In addition, cell were kept as unstimulated control. All cultures were adjusted to a final concentration of 0.21% DMSO (vehicle for TPA/A23187 plus inhibitor). ( a , c ) IL-22 and IL-6 mRNA were determined by realtime PCR. Target mRNA was normalized to GAPDH. Data are depicted as % of TPA/A23187-stimulation (means ± SD; * p

    Article Snippet: Analysis of IL-22 and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mRNA by standard PCR Total RNA, isolated by Tri-Reagent (Sigma-Aldrich) was transcribed using random hexameric primers (Qiagen, Hilden, Germany) and Moloney virus reverse transcriptase (Life Techno-logies).

    Techniques: Concentration Assay, 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

    ZEB1-AS1  downregulates  miR-200b , upregulates fascin-1 ( FSCN1 ), and promotes cell migration and invasion.  a  Luciferase assays were performed in T24, RT4, and 293 T cells co-transfected for 24 h with miR-negative control (NC) or  miR-200b  and a plasmid containing wild-type or mutant-type  ZEB1-AS1  3′untranslated region (UTR) upstream the luciferase gene. Firefly luciferase activity of each sample was normalized by Renilla luciferase activity. Data were analyzed by T-test.  b  RNA-binding protein immunoprecipitation (RIP) assays with anti-AGO2 antibodies were performed in T24 and RT4 cells transiently transfected with  miR-200b ;  ZEB1-AS1  levels were detected by quantitative PCR (qPCR); 10% input was used as positive control and RIP with anti-IgG antibodies served as negative control. Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) was used as the internal control. Data were analyzed by T-test.  c  Transfection efficiency of  ZEB1-AS1  knockdown (si1, si2) and overexpression (OE) in T24 and RT4 cells was detected by qPCR. Data were analyzed by T-test.  d  and  e  The levels of  miR-200b  ( d ) and  FSCN1  ( e ) were measured by qPCR after  ZEB1-AS1  knocked down or overexpressed in T24 and RT4 cells. Data were analyzed by T-test.  f  E-cadherin, N-cadherin, vimentin, and FSCN1 protein expression in T24 and RT4 cells in which  ZEB1-AS1  had been knocked down or overexpressed. Data were analyzed by T-test. ( g ) FSCN1 levels in T24 cells co-transfected with miR-NC or  miR-200b  and with an empty vector (EV) or a plasmid overexpressing  ZEB1-AS1 . Data were analyzed by T-test.  h  and  i  Transwell assays (without or with Matrigel) to detect cell migration ( h ) and invasion ( i ) of T24 and RT4 after  ZEB1-AS1  silencing or overexpression.  j  and  k  Transwell assays (without or with Matrigel) to detect cell migration ( j ) and invasion ( k ) of T24 and RT4 co-transfected with miR-NC or  miR-200b  and with an EV or a plasmid overexpressing  ZEB1-AS1 . Data were analyzed by T-test. Data are presented as the mean ± standard deviation (SD). * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Long non-coding RNA ZEB1-AS1 regulates miR-200b/FSCN1 signaling and enhances migration and invasion induced by TGF-β1 in bladder cancer cells

    doi: 10.1186/s13046-019-1102-6

    Figure Lengend Snippet: ZEB1-AS1 downregulates miR-200b , upregulates fascin-1 ( FSCN1 ), and promotes cell migration and invasion. a Luciferase assays were performed in T24, RT4, and 293 T cells co-transfected for 24 h with miR-negative control (NC) or miR-200b and a plasmid containing wild-type or mutant-type ZEB1-AS1 3′untranslated region (UTR) upstream the luciferase gene. Firefly luciferase activity of each sample was normalized by Renilla luciferase activity. Data were analyzed by T-test. b RNA-binding protein immunoprecipitation (RIP) assays with anti-AGO2 antibodies were performed in T24 and RT4 cells transiently transfected with miR-200b ; ZEB1-AS1 levels were detected by quantitative PCR (qPCR); 10% input was used as positive control and RIP with anti-IgG antibodies served as negative control. Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) was used as the internal control. Data were analyzed by T-test. c Transfection efficiency of ZEB1-AS1 knockdown (si1, si2) and overexpression (OE) in T24 and RT4 cells was detected by qPCR. Data were analyzed by T-test. d and e The levels of miR-200b ( d ) and FSCN1 ( e ) were measured by qPCR after ZEB1-AS1 knocked down or overexpressed in T24 and RT4 cells. Data were analyzed by T-test. f E-cadherin, N-cadherin, vimentin, and FSCN1 protein expression in T24 and RT4 cells in which ZEB1-AS1 had been knocked down or overexpressed. Data were analyzed by T-test. ( g ) FSCN1 levels in T24 cells co-transfected with miR-NC or miR-200b and with an empty vector (EV) or a plasmid overexpressing ZEB1-AS1 . Data were analyzed by T-test. h and i Transwell assays (without or with Matrigel) to detect cell migration ( h ) and invasion ( i ) of T24 and RT4 after ZEB1-AS1 silencing or overexpression. j and k Transwell assays (without or with Matrigel) to detect cell migration ( j ) and invasion ( k ) of T24 and RT4 co-transfected with miR-NC or miR-200b and with an EV or a plasmid overexpressing ZEB1-AS1 . Data were analyzed by T-test. Data are presented as the mean ± standard deviation (SD). * P

    Article Snippet: The antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH; loading control) was purchased from Sigma-Aldrich (St. Louis, MO, USA).

    Techniques: Migration, Luciferase, Transfection, Negative Control, Plasmid Preparation, Mutagenesis, Activity Assay, RNA Binding Assay, Immunoprecipitation, Real-time Polymerase Chain Reaction, Positive Control, Over Expression, Expressing, Standard Deviation

    Analysis and targeting of the plasminogen activation pathway results in decreased extracellular matrix (ECM) degradation by ductal carcinoma in situ (DCIS) structures formed in mammary architecture and microenvironment engineering (MAME) cultures. a Media conditioned from 8-day 3D cultures of myoepithelial cells (MEPs) alone, DCIS cells alone, and DCIS-MEP cocultures were analyzed by immunoblotting for plasminogen activator inhibitor 1 (PAI-1) and pro-urokinase plasminogen activator (pro-uPA). Immunoblotting for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in cell lysates was used as a loading control. b Representative images from a tissue microarray containing adjacent normal and DCIS specimens were stained for human urokinase plasminogen activator receptor (uPAR) with ATN-617 antibody (5 μg/ml) ( top rows ) and preimmune immunoglobulin G (IgG) (control). Scale bar = 100 μm ( middle rows ). In DCIS specimens imaged at a higher magnification, stromal cells ( arrows ) can be seen to exhibit strong staining for uPAR. Scale bar = 50 μm ( bottom rows ). All sections were counterstained with hematoxylin. c MCF10.DCIS (DCIS) cells were seeded in reconstituted basement membrane (rBM) overlay cultures containing dye-quenched collagen IV (DQ-collagen IV) in the absence (control) or presence of 250 nM human recombinant plasminogen activator inhibitor 1 (rPAI-1), preimmune IgG (IgG), or 10 μg/ml uPAR blocking antibody (ATN-617) and imaged live at day 4. Representative angled views of 3D reconstructions of DCIS structures illustrate nuclei ( blue ) and DQ-collagen IV degradation products (dDQ-IV, green ). One grid unit = 45 μm. d Intensity of dDQ-IV per cell was quantified from a minimum of three independent experiments using Volocity software ( n = 8–14). * p ≤ 0.05 and *** p ≤ 0.0005 as determined by unpaired t test, two-sided. MCF10.DCIS-lenti-RFP cells (DCIS, red ) were seeded into rBM overlay cultures containing DQ-collagen IV in the absence (control) or presence of rPAI-1 and imaged live at day 4. e Representative angled views of 3D reconstructions of DCIS ( red ) structures and associated dDQ-IV ( green ). One grid unit = 180 μm. f Volumes of DCIS structures ( red ) and dDQ-IV ( green ) in the absence (control) and presence of rPAI-1 were quantified from a minimum of three independent experiments using Volocity software ( n = 4). * p ≤ 0.05 and ** p ≤ 0.005 as determined by unpaired t test, two-sided. Data are presented as box-and-whisker plots where the box represents the interquartile range and whiskers represent minimum and maximum values. Images are representative of at least three independent experiments. Additional results are shown in Additional file 11 : Table S1, Additional file 12 : Table S2, and Additional file 13 : Table S3

    Journal: Breast Cancer Research : BCR

    Article Title: Pathomimetic avatars reveal divergent roles of microenvironment in invasive transition of ductal carcinoma in situ

    doi: 10.1186/s13058-017-0847-0

    Figure Lengend Snippet: Analysis and targeting of the plasminogen activation pathway results in decreased extracellular matrix (ECM) degradation by ductal carcinoma in situ (DCIS) structures formed in mammary architecture and microenvironment engineering (MAME) cultures. a Media conditioned from 8-day 3D cultures of myoepithelial cells (MEPs) alone, DCIS cells alone, and DCIS-MEP cocultures were analyzed by immunoblotting for plasminogen activator inhibitor 1 (PAI-1) and pro-urokinase plasminogen activator (pro-uPA). Immunoblotting for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in cell lysates was used as a loading control. b Representative images from a tissue microarray containing adjacent normal and DCIS specimens were stained for human urokinase plasminogen activator receptor (uPAR) with ATN-617 antibody (5 μg/ml) ( top rows ) and preimmune immunoglobulin G (IgG) (control). Scale bar = 100 μm ( middle rows ). In DCIS specimens imaged at a higher magnification, stromal cells ( arrows ) can be seen to exhibit strong staining for uPAR. Scale bar = 50 μm ( bottom rows ). All sections were counterstained with hematoxylin. c MCF10.DCIS (DCIS) cells were seeded in reconstituted basement membrane (rBM) overlay cultures containing dye-quenched collagen IV (DQ-collagen IV) in the absence (control) or presence of 250 nM human recombinant plasminogen activator inhibitor 1 (rPAI-1), preimmune IgG (IgG), or 10 μg/ml uPAR blocking antibody (ATN-617) and imaged live at day 4. Representative angled views of 3D reconstructions of DCIS structures illustrate nuclei ( blue ) and DQ-collagen IV degradation products (dDQ-IV, green ). One grid unit = 45 μm. d Intensity of dDQ-IV per cell was quantified from a minimum of three independent experiments using Volocity software ( n = 8–14). * p ≤ 0.05 and *** p ≤ 0.0005 as determined by unpaired t test, two-sided. MCF10.DCIS-lenti-RFP cells (DCIS, red ) were seeded into rBM overlay cultures containing DQ-collagen IV in the absence (control) or presence of rPAI-1 and imaged live at day 4. e Representative angled views of 3D reconstructions of DCIS ( red ) structures and associated dDQ-IV ( green ). One grid unit = 180 μm. f Volumes of DCIS structures ( red ) and dDQ-IV ( green ) in the absence (control) and presence of rPAI-1 were quantified from a minimum of three independent experiments using Volocity software ( n = 4). * p ≤ 0.05 and ** p ≤ 0.005 as determined by unpaired t test, two-sided. Data are presented as box-and-whisker plots where the box represents the interquartile range and whiskers represent minimum and maximum values. Images are representative of at least three independent experiments. Additional results are shown in Additional file 11 : Table S1, Additional file 12 : Table S2, and Additional file 13 : Table S3

    Article Snippet: Human affinity-purified IL-6 neutralizing antibody (nAb; AF-206-NA) was purchased from R & D Systems (Minneapolis, MN, USA), and anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH) was obtained from EMD Millipore (Billerica, MA, USA).

    Techniques: Activation Assay, In Situ, Microarray, Staining, Recombinant, Blocking Assay, Software, Whisker Assay

    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

    VP4 is concentrated along the nuclear envelope. (A) Cos7 cells were transiently transfected with either empty vector (−) or VP4 (+) in the presence of the tetracycline repressor. Tetracycline was added for the indicated times. Whole-cell lysates were immunoblotted with antisera against myc to detect VP4 or the loading control GAPDH. (B) Confocal immunofluorescence microscopy localization of VP4 in Cos7 cells was determined after 6 h of induction. Samples were labeled with myc (VP4-myc), ERp57, emerin, or lamin antisera and stained with either Hoechst or DAPI (DAPI, 4′,6-diamidino-2-phenylindole) as indicated. A fraction of the VP4-expressing cells displayed profound nuclear envelope disruption, as displayed in the lamin lower panel. Bars, 10 μm.

    Journal: Journal of Virology

    Article Title: The Simian Virus 40 Late Viral Protein VP4 Disrupts the Nuclear Envelope for Viral Release

    doi: 10.1128/JVI.07047-11

    Figure Lengend Snippet: VP4 is concentrated along the nuclear envelope. (A) Cos7 cells were transiently transfected with either empty vector (−) or VP4 (+) in the presence of the tetracycline repressor. Tetracycline was added for the indicated times. Whole-cell lysates were immunoblotted with antisera against myc to detect VP4 or the loading control GAPDH. (B) Confocal immunofluorescence microscopy localization of VP4 in Cos7 cells was determined after 6 h of induction. Samples were labeled with myc (VP4-myc), ERp57, emerin, or lamin antisera and stained with either Hoechst or DAPI (DAPI, 4′,6-diamidino-2-phenylindole) as indicated. A fraction of the VP4-expressing cells displayed profound nuclear envelope disruption, as displayed in the lamin lower panel. Bars, 10 μm.

    Article Snippet: The following antibodies were obtained as indicated: myc, lamin A/C, and emerin (Cell Signaling, Danvers, MA); GAPDH (glyceraldehyde-3-phosphate dehydrogenase; Millipore, Billerica, MA); VP1 and VP2/3 (Abcam, Cambridge, MA; VP2/3 was also obtained from A. Oppenheim, Jerusalem, Israel); calnexin (Enzo Life Sciences Inc., Ann Arbor, MI); large T antigen (LT; Merck KGaA, Darmstadt, Germany); and glutathione S -transferase (GST; Abmart, Arlington, MA).

    Techniques: Transfection, Plasmid Preparation, Immunofluorescence, Microscopy, Labeling, Staining, Expressing

    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

    Expression of Nrdp1 and cell-cycle markers in proliferating HCC cells. ( A ) HepG2 cells were synchronized by serum starvation for 72 h. Upon serum release, cell lysates were prepared and analyzed by Western blotting using antibodies directed against Nrdp1, PCNA, and Cyclin D1. GAPDH was used as a control for protein loading and integrity. ( B ) Bar chart shows the ratio of Nrdp1, PCNA, and Cyclin D1 to GAPDH for each time point as measured by densitometry. S, serum starvation; R, serum release. ( C ) Cells were synchronized at G1 after serum starvation for 72 h, then allowed to progress through the cell cycle by adding medium containing 10% FBS for the indicated times (R4 h, R8 h, R12 h, and R24 h). Data are shown as mean ± SD for three experiments. Abbreviations: Nrdp1, neuregulin receptor degradation protein-1; HCC, hepatocellular carcinoma; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PCNA, proliferating cell nuclear antigen; FBS, fetal bovine serum.

    Journal: OncoTargets and therapy

    Article Title: Reduced expression of Nrdp1 predicts a poor prognosis in human hepatocellular carcinoma

    doi: 10.2147/OTT.S160638

    Figure Lengend Snippet: Expression of Nrdp1 and cell-cycle markers in proliferating HCC cells. ( A ) HepG2 cells were synchronized by serum starvation for 72 h. Upon serum release, cell lysates were prepared and analyzed by Western blotting using antibodies directed against Nrdp1, PCNA, and Cyclin D1. GAPDH was used as a control for protein loading and integrity. ( B ) Bar chart shows the ratio of Nrdp1, PCNA, and Cyclin D1 to GAPDH for each time point as measured by densitometry. S, serum starvation; R, serum release. ( C ) Cells were synchronized at G1 after serum starvation for 72 h, then allowed to progress through the cell cycle by adding medium containing 10% FBS for the indicated times (R4 h, R8 h, R12 h, and R24 h). Data are shown as mean ± SD for three experiments. Abbreviations: Nrdp1, neuregulin receptor degradation protein-1; HCC, hepatocellular carcinoma; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PCNA, proliferating cell nuclear antigen; FBS, fetal bovine serum.

    Article Snippet: Membranes were washed, blocked, and incubated with anti-Nrdp1 antibody (1:1,000; Santa Cruz Biotechnology, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (1:1,000; Sigma).

    Techniques: Expressing, Western Blot

    Knockdown of Nrdp1 suppresses cell proliferation and altered cell-cycle progression in L02 cells. ( A ) Western blotting shows that the expression of Nrdp1, Cyclin D1, and PCNA decline in L02 cells treated with siRNA targeting Nrdp1, compared with a negative control and mock siRNA treatments. ( B ) Bar chart shows the ratio of Nrdp1, Cyclin D1, and PCNA to GAPDH as measured by densitometry. ( C ) The CCK-8 assay was used to measure cell proliferation. L02 cells treated with siNrdp1-1 and siNrdp1-3 exhibit significantly enhanced proliferation. Absorbance was used to examine proliferation at each indicated time (0, 1, 2, and 3 days). ( D ) Cell-cycle analysis shows that knockdown of Nrdp1 by siNrdp1-1 and siNrdp1-3 delays the G1–S transition and arrests cells in the G1 phase of L02 cells, as shown by flow cytometry. Data are shown as mean ± SD for three experiments. Abbreviations: Nrdp1, neuregulin receptor degradation protein-1; CON, negative control; Mock, mock siRNAtreatments; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PCNA, proliferating cell nuclear antigen.

    Journal: OncoTargets and therapy

    Article Title: Reduced expression of Nrdp1 predicts a poor prognosis in human hepatocellular carcinoma

    doi: 10.2147/OTT.S160638

    Figure Lengend Snippet: Knockdown of Nrdp1 suppresses cell proliferation and altered cell-cycle progression in L02 cells. ( A ) Western blotting shows that the expression of Nrdp1, Cyclin D1, and PCNA decline in L02 cells treated with siRNA targeting Nrdp1, compared with a negative control and mock siRNA treatments. ( B ) Bar chart shows the ratio of Nrdp1, Cyclin D1, and PCNA to GAPDH as measured by densitometry. ( C ) The CCK-8 assay was used to measure cell proliferation. L02 cells treated with siNrdp1-1 and siNrdp1-3 exhibit significantly enhanced proliferation. Absorbance was used to examine proliferation at each indicated time (0, 1, 2, and 3 days). ( D ) Cell-cycle analysis shows that knockdown of Nrdp1 by siNrdp1-1 and siNrdp1-3 delays the G1–S transition and arrests cells in the G1 phase of L02 cells, as shown by flow cytometry. Data are shown as mean ± SD for three experiments. Abbreviations: Nrdp1, neuregulin receptor degradation protein-1; CON, negative control; Mock, mock siRNAtreatments; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PCNA, proliferating cell nuclear antigen.

    Article Snippet: Membranes were washed, blocked, and incubated with anti-Nrdp1 antibody (1:1,000; Santa Cruz Biotechnology, CA, USA) and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (1:1,000; Sigma).

    Techniques: Western Blot, Expressing, Negative Control, CCK-8 Assay, Cell Cycle Assay, Flow Cytometry, Cytometry

    Methylation of CpG islands of ER stress gene promoters in the livers of mice at different ages. Grp94, glucose-regulated protein 94; Gapdh, glyceraldehyde-3-phosphate dehydrogenase; Chop, DNA damage-inducible transcript 3, also known as C/EBP homologous protein; Atf6, activating transcription factor 6; PDI, protein disulfite isomerase; MW, wild type of middle age; OW, wild type of older age; MK, BiP knockout of middle age; OK, BiP knockout of older age without liver tumors; OKL, normal liver portion from tumor bearing livers of older BiP knockouts; OKT, tumor portion from tumor bearing livers of older BiP knockouts; * p

    Journal: Frontiers in Genetics

    Article Title: Altered methylation and expression of ER-associated degradation factors in long-term alcohol and constitutive ER stress-induced murine hepatic tumors

    doi: 10.3389/fgene.2013.00224

    Figure Lengend Snippet: Methylation of CpG islands of ER stress gene promoters in the livers of mice at different ages. Grp94, glucose-regulated protein 94; Gapdh, glyceraldehyde-3-phosphate dehydrogenase; Chop, DNA damage-inducible transcript 3, also known as C/EBP homologous protein; Atf6, activating transcription factor 6; PDI, protein disulfite isomerase; MW, wild type of middle age; OW, wild type of older age; MK, BiP knockout of middle age; OK, BiP knockout of older age without liver tumors; OKL, normal liver portion from tumor bearing livers of older BiP knockouts; OKT, tumor portion from tumor bearing livers of older BiP knockouts; * p

    Article Snippet: Primary antibodies against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were from Millipore (Billerica, MA, USA).

    Techniques: Methylation, Mouse Assay, Knock-Out

    Prostaglandin E 2 (PGE 2 ) increases α7 nicotinic acetylcholine receptor (nAChR) protein expression and induces cell growth partly through α7 nAChR-dependent cholinergic signaling. (a) Cellular protein was isolated from H1792 cells that were cultured with increased concentrations of dmPGE 2 as indicated for 24 hours followed by Western blot analysis with antibodies against α7 nAChR protein. (b) Cellular protein was isolated from H1792 cells that were cultured with dmPGE 2 (1 μM) for the indicated time, followed by Western blot analysis with antibodies against α7 nAChR protein. (c) Cellular protein was isolated from several non-small cell lung cancer (NSCLC) cell lines (A549, H1792, H1838, and H358) that were cultured with dmPGE 2 (0.1, 1 μM) for up to 24 hours, followed by Western blot analysis with antibodies against α7 nAChR protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as an internal control for normalization purposes. (d) Total ribonucleic acid (RNA) was isolated from several NSCLC cell lines (A549, H1792, H1838, and H358) treated with dmPGE 2 (0.1, 1 μM) for up to 24 hours and real time reverse transcription polymerase chain reaction was performed for evaluating α7 nAChR messenger RNA expression. GAPDH served as an internal control for normalization purposes. (e) H1838 cells were transfected with control or α7 nAChR small interfering RNA (100 nM) for 24 hours before exposure of the cells to dmPGE 2 (0.1 μM) for up to three days. The viable cells were then determined by Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega). (f) H1792 and H1838 cells were cultured with dmPGE 2 (0.1 μM) in the presence or absence of AChE (50 U/mL) for up to five days. The viable cells were then detected using the Cell Titer-Glo Luminescent Cell Viability Assay kit. All data were depicted as mean ± standard deviation. * indicates significant difference compared to the untreated cell group. ** indicates significant difference of combination treatment compared to dmPGE 2 alone ( P

    Journal: Thoracic Cancer

    Article Title: Novel link between prostaglandin E2 (PGE2) and cholinergic signaling in lung cancer: The role of c-Jun in PGE2-induced α7 nicotinic acetylcholine receptor expression and tumor cell proliferation

    doi: 10.1111/1759-7714.12219

    Figure Lengend Snippet: Prostaglandin E 2 (PGE 2 ) increases α7 nicotinic acetylcholine receptor (nAChR) protein expression and induces cell growth partly through α7 nAChR-dependent cholinergic signaling. (a) Cellular protein was isolated from H1792 cells that were cultured with increased concentrations of dmPGE 2 as indicated for 24 hours followed by Western blot analysis with antibodies against α7 nAChR protein. (b) Cellular protein was isolated from H1792 cells that were cultured with dmPGE 2 (1 μM) for the indicated time, followed by Western blot analysis with antibodies against α7 nAChR protein. (c) Cellular protein was isolated from several non-small cell lung cancer (NSCLC) cell lines (A549, H1792, H1838, and H358) that were cultured with dmPGE 2 (0.1, 1 μM) for up to 24 hours, followed by Western blot analysis with antibodies against α7 nAChR protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as an internal control for normalization purposes. (d) Total ribonucleic acid (RNA) was isolated from several NSCLC cell lines (A549, H1792, H1838, and H358) treated with dmPGE 2 (0.1, 1 μM) for up to 24 hours and real time reverse transcription polymerase chain reaction was performed for evaluating α7 nAChR messenger RNA expression. GAPDH served as an internal control for normalization purposes. (e) H1838 cells were transfected with control or α7 nAChR small interfering RNA (100 nM) for 24 hours before exposure of the cells to dmPGE 2 (0.1 μM) for up to three days. The viable cells were then determined by Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega). (f) H1792 and H1838 cells were cultured with dmPGE 2 (0.1 μM) in the presence or absence of AChE (50 U/mL) for up to five days. The viable cells were then detected using the Cell Titer-Glo Luminescent Cell Viability Assay kit. All data were depicted as mean ± standard deviation. * indicates significant difference compared to the untreated cell group. ** indicates significant difference of combination treatment compared to dmPGE 2 alone ( P

    Article Snippet: Acetylcholinesterase (AChE), antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and other chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA) unless otherwise indicated.

    Techniques: Expressing, Isolation, Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, RNA Expression, Transfection, Small Interfering RNA, Cell Viability Assay, Standard Deviation

    The role of c-Jun in mediating the effect of prostaglandin E 2 (PGE 2 ) on α7 nicotinic acetylcholine receptor (nAChR) expression and cell growth. (a) H1792 cells were transfected with control or c-Jun small interfering ribonucleic acid (siRNA) (100 nM) for 24 hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours. Western blot analysis was then performed to examine for α7 nAChR and c-Jun protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as loading control. (b) H1792 cells were transfected with control or c-Jun siRNAs (100 nM) together with a wild type Chrna7 promoter reporter construct ligated to a luciferase reporter gene and an internal control for 24 hours. Cells were then exposed to dmPGE 2 (1 μM) for an additional 24 hours. A Dual Luciferase Reporter kit (Promega) determined luciferase activity. The insert in the upper panel represents Western blot results for c-Jun protein. GAPDH served as internal control for normalization purposes. (c) H1838 cells were transfected with control or c-Jun siRNAs (100 nM) for 24 hours. Cells were then exposed to dmPGE 2 (0.1, 1 μM) for an additional 72 hours. A Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega) determined cell numbers. The insert in the upper panel represents Western blot results for c-Jun protein. GAPDH served as internal control for normalization purposes. The bars below represent the mean ± standard deviation of at least three independent experiments for each condition. * indicates significant increase of activity compared to controls. ** indicates significance of combination treatment compared to dmPGE 2 alone ( P

    Journal: Thoracic Cancer

    Article Title: Novel link between prostaglandin E2 (PGE2) and cholinergic signaling in lung cancer: The role of c-Jun in PGE2-induced α7 nicotinic acetylcholine receptor expression and tumor cell proliferation

    doi: 10.1111/1759-7714.12219

    Figure Lengend Snippet: The role of c-Jun in mediating the effect of prostaglandin E 2 (PGE 2 ) on α7 nicotinic acetylcholine receptor (nAChR) expression and cell growth. (a) H1792 cells were transfected with control or c-Jun small interfering ribonucleic acid (siRNA) (100 nM) for 24 hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours. Western blot analysis was then performed to examine for α7 nAChR and c-Jun protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as loading control. (b) H1792 cells were transfected with control or c-Jun siRNAs (100 nM) together with a wild type Chrna7 promoter reporter construct ligated to a luciferase reporter gene and an internal control for 24 hours. Cells were then exposed to dmPGE 2 (1 μM) for an additional 24 hours. A Dual Luciferase Reporter kit (Promega) determined luciferase activity. The insert in the upper panel represents Western blot results for c-Jun protein. GAPDH served as internal control for normalization purposes. (c) H1838 cells were transfected with control or c-Jun siRNAs (100 nM) for 24 hours. Cells were then exposed to dmPGE 2 (0.1, 1 μM) for an additional 72 hours. A Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega) determined cell numbers. The insert in the upper panel represents Western blot results for c-Jun protein. GAPDH served as internal control for normalization purposes. The bars below represent the mean ± standard deviation of at least three independent experiments for each condition. * indicates significant increase of activity compared to controls. ** indicates significance of combination treatment compared to dmPGE 2 alone ( P

    Article Snippet: Acetylcholinesterase (AChE), antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and other chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA) unless otherwise indicated.

    Techniques: Expressing, Transfection, Western Blot, Construct, Luciferase, Activity Assay, Cell Viability Assay, Standard Deviation

    Prostaglandin E 2 (PGE 2 ) induces c-Jun protein expression via phosphatidylinositol 3-kinase (PI3-K), protein kinase A (PKA), and c-Jun N-terminal kinase (JNK) signals. (a) Cellular proteins were isolated from H1792 cells treated with increased concentrations of dmPGE 2 for 24 hours. Afterwards, Western Blot analyses were performed using a polyclonal antibody against c-Jun protein. (b–d) Cellular protein was isolated from H1792 cells treated with H-89 (10 μM), PD98059 (10 μM), Wortmannin (1 μM), LY294002 (10 μM), or JNK inhibitor II (20 μM) for two hours before exposure of the cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western blot analysis for c-Jun protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as internal control for loading purposes. Control , indicates untreated control cells.

    Journal: Thoracic Cancer

    Article Title: Novel link between prostaglandin E2 (PGE2) and cholinergic signaling in lung cancer: The role of c-Jun in PGE2-induced α7 nicotinic acetylcholine receptor expression and tumor cell proliferation

    doi: 10.1111/1759-7714.12219

    Figure Lengend Snippet: Prostaglandin E 2 (PGE 2 ) induces c-Jun protein expression via phosphatidylinositol 3-kinase (PI3-K), protein kinase A (PKA), and c-Jun N-terminal kinase (JNK) signals. (a) Cellular proteins were isolated from H1792 cells treated with increased concentrations of dmPGE 2 for 24 hours. Afterwards, Western Blot analyses were performed using a polyclonal antibody against c-Jun protein. (b–d) Cellular protein was isolated from H1792 cells treated with H-89 (10 μM), PD98059 (10 μM), Wortmannin (1 μM), LY294002 (10 μM), or JNK inhibitor II (20 μM) for two hours before exposure of the cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western blot analysis for c-Jun protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as internal control for loading purposes. Control , indicates untreated control cells.

    Article Snippet: Acetylcholinesterase (AChE), antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and other chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA) unless otherwise indicated.

    Techniques: Expressing, Isolation, Western Blot

    Prostaglandin E 2 (PGE 2 ) increases α7 nicotinic acetylcholine receptor (nAChR) gene expression via EP4 and phosphatidylinositol 3-kinase (PI3-K), protein kinase A (PKA), and c-Jun N-terminal kinase (JNK) signals. (a) Cellular protein was isolated from H1792 cells cultured for up to two hours in the presence or absence of AH23848 (5 μM) before exposure of cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western blot analysis for α7 nAChR protein. (b) Cellular protein was isolated from H1792 cells cultured for 24 hours in the presence or absence of the control or EP4 small interfering ribonucleic acid (100 nM) before exposure of cells to dmPGE 2 (1 μM) for an additional 24 hours, and then subjected to Western blot analysis for EP4 and α7 nAChR protein. (c) H1792 and H1838 cells were cultured with dmPGE 2 (0.1 μM) in the presence or absence of AH23848 (5 μM) for up to five days. The viable cells were then detected using a Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega). (d) Cellular protein was isolated from H1838 and H1792 cells treated with dmPGE 2 (1 μM) for the indicated time followed by Western blot analysis with antibodies against phospho-SAPK/JNK and SAPK/JNK proteins. (e) Cellular protein was isolated from H1792 cells treated with H-89 (10 μM) or PD98059 (10 μM) for two hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. (f) Cellular protein was isolated from H1792 cells treated with JNK inhibitor II (20 μM) for two hours before exposure of the cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. (g) Cellular protein was isolated from H1792 cells treated with Wortmannin (1 μM) or LY294002 (10 μM) for two hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as internal control for loading purposes. Control , indicates untreated control cells. * indicates significant difference compared to the untreated cell group. ** indicates significant difference of combination treatment as compared to the dmPGE 2 alone ( P

    Journal: Thoracic Cancer

    Article Title: Novel link between prostaglandin E2 (PGE2) and cholinergic signaling in lung cancer: The role of c-Jun in PGE2-induced α7 nicotinic acetylcholine receptor expression and tumor cell proliferation

    doi: 10.1111/1759-7714.12219

    Figure Lengend Snippet: Prostaglandin E 2 (PGE 2 ) increases α7 nicotinic acetylcholine receptor (nAChR) gene expression via EP4 and phosphatidylinositol 3-kinase (PI3-K), protein kinase A (PKA), and c-Jun N-terminal kinase (JNK) signals. (a) Cellular protein was isolated from H1792 cells cultured for up to two hours in the presence or absence of AH23848 (5 μM) before exposure of cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western blot analysis for α7 nAChR protein. (b) Cellular protein was isolated from H1792 cells cultured for 24 hours in the presence or absence of the control or EP4 small interfering ribonucleic acid (100 nM) before exposure of cells to dmPGE 2 (1 μM) for an additional 24 hours, and then subjected to Western blot analysis for EP4 and α7 nAChR protein. (c) H1792 and H1838 cells were cultured with dmPGE 2 (0.1 μM) in the presence or absence of AH23848 (5 μM) for up to five days. The viable cells were then detected using a Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega). (d) Cellular protein was isolated from H1838 and H1792 cells treated with dmPGE 2 (1 μM) for the indicated time followed by Western blot analysis with antibodies against phospho-SAPK/JNK and SAPK/JNK proteins. (e) Cellular protein was isolated from H1792 cells treated with H-89 (10 μM) or PD98059 (10 μM) for two hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. (f) Cellular protein was isolated from H1792 cells treated with JNK inhibitor II (20 μM) for two hours before exposure of the cells to dmPGE 2 (0.1, 1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. (g) Cellular protein was isolated from H1792 cells treated with Wortmannin (1 μM) or LY294002 (10 μM) for two hours before exposure of the cells to dmPGE 2 (1 μM) for an additional 48 hours, then subjected to Western Blot analysis for α7 nAChR protein. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) served as internal control for loading purposes. Control , indicates untreated control cells. * indicates significant difference compared to the untreated cell group. ** indicates significant difference of combination treatment as compared to the dmPGE 2 alone ( P

    Article Snippet: Acetylcholinesterase (AChE), antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and other chemicals were purchased from Sigma Aldrich (St. Louis, MO, USA) unless otherwise indicated.

    Techniques: Expressing, Isolation, Cell Culture, Western Blot, Cell Viability Assay

    Expression of Zp in rodent tissues ( A , B ) and human cell lines ( C ). ( A ) Expression of Zp mRNA in E18 mouse placenta, liver, heart, kidney, E7 placenta, E18 embryo, and enterocytes. GAPDH expression was used as a loading control. ( B ) Immunoblot with Zp-specific IgG of rat placenta, lactating mouse mammary tissue, mouse E18 embryo, and ( C ). mouse serum and enterocytes, and human BeWo, MCF7, T47D, and MCF10AT cell line extracts. ( D ) Immunoblot with Cp- and Heph-specific IgG, respectively, of mouse serum and enterocytes; 50 μ g total protein/lane.

    Journal: The Journal of Nutrition

    Article Title: Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2 3

    doi: 10.3945/jn.109.117531

    Figure Lengend Snippet: Expression of Zp in rodent tissues ( A , B ) and human cell lines ( C ). ( A ) Expression of Zp mRNA in E18 mouse placenta, liver, heart, kidney, E7 placenta, E18 embryo, and enterocytes. GAPDH expression was used as a loading control. ( B ) Immunoblot with Zp-specific IgG of rat placenta, lactating mouse mammary tissue, mouse E18 embryo, and ( C ). mouse serum and enterocytes, and human BeWo, MCF7, T47D, and MCF10AT cell line extracts. ( D ) Immunoblot with Cp- and Heph-specific IgG, respectively, of mouse serum and enterocytes; 50 μ g total protein/lane.

    Article Snippet: Cp-specific IgG was from Accurate Chemical and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) IgG was from Chemicon.

    Techniques: Expressing

    pPD oxidase activity in BeWo cells after siRNA knockdown of Zp. ( A ) Immunoblot with Zp- and GAPDH-specific IgG of extracts (100 μ g total protein/lane) of nontransfected BeWo cells (Ctrl) and BeWo cells transfected with set 1, set 2, and set 3 siRNA targeting Zp. Purified human Cp was used as a control (20 μ g/lane). ( B ) In-gel pPD activity assay of the same samples (100 μ g total protein/lane).

    Journal: The Journal of Nutrition

    Article Title: Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2 3

    doi: 10.3945/jn.109.117531

    Figure Lengend Snippet: pPD oxidase activity in BeWo cells after siRNA knockdown of Zp. ( A ) Immunoblot with Zp- and GAPDH-specific IgG of extracts (100 μ g total protein/lane) of nontransfected BeWo cells (Ctrl) and BeWo cells transfected with set 1, set 2, and set 3 siRNA targeting Zp. Purified human Cp was used as a control (20 μ g/lane). ( B ) In-gel pPD activity assay of the same samples (100 μ g total protein/lane).

    Article Snippet: Cp-specific IgG was from Accurate Chemical and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) IgG was from Chemicon.

    Techniques: Activity Assay, Transfection, Purification

    Regulation of Zp by copper in BeWo cells. ( A ) BeWo cells were made copper-deficient by incubation with increasing concentrations of BCS, and cell extracts (100 μ g total protein/lane) were then immunoblotted with Zp-specific and GAPDH-specific IgG followed by development with chemiluminescence and measurement of band intensity by densitometry. The ratio of each Zp band density to the density of the 0 μ mol/L BCS control were multiplied by 100, averaged, and then plotted versus BCS concentration ( r 2 = 0.926; P ≤ 0.0005). Values are means ± SD of results from 3 independent experiments. ( B ) SOD1 activity as measured in the same cell lysates as in A and plotted versus BCS concentration ( r 2 = 0.982; P ≤ 0.0001). Values are means ± SD, n = 2 independent experiments.

    Journal: The Journal of Nutrition

    Article Title: Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2Identification of Zyklopen, a New Member of the Vertebrate Multicopper Ferroxidase Family, and Characterization in Rodents and Human Cells 1 2 3

    doi: 10.3945/jn.109.117531

    Figure Lengend Snippet: Regulation of Zp by copper in BeWo cells. ( A ) BeWo cells were made copper-deficient by incubation with increasing concentrations of BCS, and cell extracts (100 μ g total protein/lane) were then immunoblotted with Zp-specific and GAPDH-specific IgG followed by development with chemiluminescence and measurement of band intensity by densitometry. The ratio of each Zp band density to the density of the 0 μ mol/L BCS control were multiplied by 100, averaged, and then plotted versus BCS concentration ( r 2 = 0.926; P ≤ 0.0005). Values are means ± SD of results from 3 independent experiments. ( B ) SOD1 activity as measured in the same cell lysates as in A and plotted versus BCS concentration ( r 2 = 0.982; P ≤ 0.0001). Values are means ± SD, n = 2 independent experiments.

    Article Snippet: Cp-specific IgG was from Accurate Chemical and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) IgG was from Chemicon.

    Techniques: Incubation, Concentration Assay, Activity Assay

    Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related glyceraldehyde-3-phosphate dehydrogenase. The results are represented as means ± SEM ( n = 4). # P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Scutellarin Exerts Hypoglycemic and Renal Protective Effects in db/db Mice via the Nrf2/HO-1 Signaling Pathway

    doi: 10.1155/2019/1354345

    Figure Lengend Snippet: Four weeks of SCU treatment increased the expression levels of Nrf2, HO-1, SOD1, SOD2, and CAT in the kidney of db/db mice. The data on quantified protein expressions were normalized by related glyceraldehyde-3-phosphate dehydrogenase. The results are represented as means ± SEM ( n = 4). # P

    Article Snippet: Then, the bands were incubated overnight at 4°C in a corresponding primary antibody solution containing Nrf2 (ab137550), HO-1 (ab13248), SOD1 (ab13498), SOD2 (ab13533), and CAT (ab16731) (1 : 2000; Abcam, UK) or glyceraldehyde 3-phosphate dehydrogenase (GAPDH, ABS16, 1 : 2000, Merck Millipore, USA) and then incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (bs-0295G, 1 : 2000, Beijing Biosynthesis Biotechnology Co. Ltd., China) for 4 hours at 4°C.

    Techniques: Expressing, Mouse Assay

    PIO interacts with and inhibits purified GAPDH. (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.

    Journal: EBioMedicine

    Article Title: Paradoxical Inhibition of Glycolysis by Pioglitazone Opposes the Mitochondriopathy Caused by AIF Deficiency

    doi: 10.1016/j.ebiom.2017.02.013

    Figure Lengend Snippet: PIO interacts with and inhibits purified GAPDH. (A) Western blot analysis of purified GAPDH in the absence (left panel) or presence (right panel) of added dithiothreitol (DTT). Noticeably, no higher aggregation forms were detected in the stacking gel. (B) Quantities of different aggregation forms (tetra-, tri-, di-, and monomeric) in the absence of DTT (open bars: initial GAPDH suspension, GAPDH plus DMSO, GAPDH plus DMSO-solubilized PIO, or in the presence of DTT (green filled bars) added before or after PIO). (C) GAPDH (2.1 mIU/ml) fluorescence in the absence (blue dots) or presence (green dots) of 12.5 μM PIO against UV (280 nm) irradiation. The presence of 1.25‰ DMSO had per se a partial protecting effect against UV (grey dots). No changes in fluorescence were observed when the enzyme was kept away from UV irradiation (black dots). Noticeably initial fluorescence in the presence or absence of PIO, or DMSO was not different ruling out potential interference of these compounds with fluorescence assay conditions (excitation: 280 nm, 10 nm bandpass; emission: 330 nm, 10 nm bandpass). (D) Initial activity of rabbit skeletal muscle GAPDH (0.05 mIU/ml) measured in the forward direction (NADH accumulation) ( Velick, 1955 ) and of increasing amounts of PIO. (E) Activity measured in the backward direction (NADH oxidation) in the absence (blue open bar) or presence (green open bar) of 200 μM PIO, and in the presence of 1.7 mM cysteine and absence (blue filled bar) or presence (green filled bar) of 200 μM PIO. (F) Lineweaver-Burk plots of forward GAPDH activity (0.075 mIU/ml) in the absence (black line) or presence (green line) of 62.5 μM PIO. GAP, d -glyceraldehyde 3-phosphate.

    Article Snippet: 2.10 Purified GAPDH Studies The activity of purified rabbit skeletal muscle NAD+ -dependent GAPDH (EC 1.2.1.12; G2267 Sigma) was spectrophotometrically measured (Cary 50 spectrophotometer; 340 minus 380 nm) in the forward direction (NADH accumulation) in 30 mM pyrophosphate buffer (pH 8.4), 0.3 mM NAD+ , using 10 mM GAP as a substrate ( ).

    Techniques: Purification, Western Blot, Fluorescence, Irradiation, Activity Assay

    Quantification of ( A ) FAK and ( B ) phospho-FAK expression evaluated by Western blot (WB), with normalization to glyceraldehyde 3-phosphate dehydrogenase (GAPDH ) expression, in nine normal lungs, 30 non-small-cell lung cancer (NSCLC), and 10 small-cell lung cancer (SCLC) tissue lysates. Each dot represents one sample. Data presented as the mean ± S.D. Significance determined by the Kruskal-Wallis test. ( C ) Illustration of a representative WB of FAK and phospho-FAK (Y397) expression in normal lung, NSCLC, and SCLC tissue lysates. All the WB are represented in Supplementary Figure S1 .

    Journal: Cancers

    Article Title: Increased Expression and Activation of FAK in Small-Cell Lung Cancer Compared to Non-Small-Cell Lung Cancer

    doi: 10.3390/cancers11101526

    Figure Lengend Snippet: Quantification of ( A ) FAK and ( B ) phospho-FAK expression evaluated by Western blot (WB), with normalization to glyceraldehyde 3-phosphate dehydrogenase (GAPDH ) expression, in nine normal lungs, 30 non-small-cell lung cancer (NSCLC), and 10 small-cell lung cancer (SCLC) tissue lysates. Each dot represents one sample. Data presented as the mean ± S.D. Significance determined by the Kruskal-Wallis test. ( C ) Illustration of a representative WB of FAK and phospho-FAK (Y397) expression in normal lung, NSCLC, and SCLC tissue lysates. All the WB are represented in Supplementary Figure S1 .

    Article Snippet: After blocking 1 h with 5% W/V BSA (Sigma, Saint-Louis, MO, USA) in TBS with 0.1% Tween 20 (Sigma), the membrane was incubated overnight at 4 °C with phospho-FAK Y397 rabbit antibody (1/1000 Cell Signaling Technology, Danvers, MA, USA) or total FAK mouse antibody (1/250, Santa Cruz Biotechnology, Dallas, TX, USA) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) rabbit antibody (1/5000, Sigma).

    Techniques: Expressing, Western Blot

    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

    Expression of TMEM106B in different cell types. ( A ) Cell lysates from HEK293T, NSC-34 and BV-2 were loaded and blotted with anti-TMEM106B and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies. ( B ) TMEM106B forms homodimers. FLAG-tagged

    Journal: Human Molecular Genetics

    Article Title: The frontotemporal lobar degeneration risk factor, TMEM106B, regulates lysosomal morphology and function

    doi: 10.1093/hmg/dds475

    Figure Lengend Snippet: Expression of TMEM106B in different cell types. ( A ) Cell lysates from HEK293T, NSC-34 and BV-2 were loaded and blotted with anti-TMEM106B and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies. ( B ) TMEM106B forms homodimers. FLAG-tagged

    Article Snippet: The following antibodies were used in the study: sheep anti-mouse PGRN antibodies, goat anti-human PGRN antibodies and goat anti-mouse sortilin from R & D systems; mouse anti-myc (9E10) and anti-FLAG (M2) antibodies from Sigma-Aldrich; mouse anti-LAMP1 and EEA1 antibodies from BD biosciences; rabbit anti-EGFR, rabbit anti-phospho-Erk1/2 and rabbit anti-cleaved caspase 3 antibodies from Cell Signaling; rat anti-mouse LAMP1 (1D4B) antibodies from BioLegend; rabbit anti-TDP 43 antibodies from Proteintech; and mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies from Millipore.

    Techniques: Expressing

    Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p

    Journal: Scientific Reports

    Article Title: Critical role of sigma-1 receptors in central neuropathic pain-related behaviours after mild spinal cord injury in mice

    doi: 10.1038/s41598-018-22217-9

    Figure Lengend Snippet: Spinal Tyr1472 and Ser1303 phosphorylation of N-methyl-D-aspartate (NMDA) receptor NR2B subunit at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. ( A ) Quantification and representative immunoblots of total NR2B, pS1303NR2B and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( B ) Quantification and representative immunoblots of total NR2B, pY1472NR2B and GAPDH. a–b: groups not sharing a letter are significantly different, p

    Article Snippet: To ensure equal protein loading, rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (1:40,000, Sigma-Aldrich Quimica S.A.) was used as a loading control.

    Techniques: Knock-Out, Mouse Assay, Western Blot

    Spinal ERK1/2 phosphorylation (pERK) expression at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. Quantification and representative immunoblots of total ERK, pERK and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Protein expressions were normalized to GAPDH and data is presented as a percentage respect to WT naïve or KO naïve mice (mean ± standard error of the mean; n = 5–6). a–b: groups not sharing a letter are significantly different, p

    Journal: Scientific Reports

    Article Title: Critical role of sigma-1 receptors in central neuropathic pain-related behaviours after mild spinal cord injury in mice

    doi: 10.1038/s41598-018-22217-9

    Figure Lengend Snippet: Spinal ERK1/2 phosphorylation (pERK) expression at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor (σ1R) knockout (KO) mice. Quantification and representative immunoblots of total ERK, pERK and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Protein expressions were normalized to GAPDH and data is presented as a percentage respect to WT naïve or KO naïve mice (mean ± standard error of the mean; n = 5–6). a–b: groups not sharing a letter are significantly different, p

    Article Snippet: To ensure equal protein loading, rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (1:40,000, Sigma-Aldrich Quimica S.A.) was used as a loading control.

    Techniques: Expressing, Knock-Out, Mouse Assay, Western Blot

    Spinal inflammatory cytokines (tumour necrosis factor [TNF]-α and interleukin[IL]-1β) expression at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor knockout (KO). ( A ) Quantification and representative immunoblots of TNF-α and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). a–b: groups not sharing a letter are significantly different, p

    Journal: Scientific Reports

    Article Title: Critical role of sigma-1 receptors in central neuropathic pain-related behaviours after mild spinal cord injury in mice

    doi: 10.1038/s41598-018-22217-9

    Figure Lengend Snippet: Spinal inflammatory cytokines (tumour necrosis factor [TNF]-α and interleukin[IL]-1β) expression at day 28 after spinal cord injury (SCI) in wild type (WT) and sigma-1 receptor knockout (KO). ( A ) Quantification and representative immunoblots of TNF-α and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). a–b: groups not sharing a letter are significantly different, p

    Article Snippet: To ensure equal protein loading, rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (1:40,000, Sigma-Aldrich Quimica S.A.) was used as a loading control.

    Techniques: Expressing, Knock-Out, Western Blot

    AICAR-dependent growth suppression and apoptosis in renal cell carcinoma cells. (A) 786-0 cells were treated for 4 days with solvent control or with the indicated concentrations of AICAR. Cell proliferation was assessed by MTT. Data are expressed as % control-treated cells and represent means ± SE of three independent experiments. Paired t-test analysis for the growth of 786-0 cells treated with 250 µmol/L AICAR, 500 µmol/L AICAR and 1,000 µmol/L AICAR versus control treated cells showed a 2-tailed p values of 0.003, 0.0001 and 0.004, respectively. (B) Top, 786-0 cells were treated with solvent control or AICAR (2 mmol/L) for 48 hours, as indicated. Equal amounts of total cell lysates were resolved by SDS-PAGE and immunoblotted with an antibody against the phosphorylated/activated form of AMPKα at Thr 172. Middle, equal protein aliquots from the same experiment were resolved on a separate gel and probed with an anti-AMPKα antibody. Bottom, the top blot was subsequently reprobed with an antibody to detect anti-glyceradledhyde-3-phosphate dehydrogenase (anti-GAPDH). (C) Top, 786-0 cells were treated with solvent control or the indicated concentrations of AICAR for 48 hours. Equal amounts of total cell lysates were resolved by SDS-PAGE and immunoblotted with an antibody against the phosphorylated/activated form of AMPKα at Thr 172. Middle, bottom, the blot was subsequently stripped and reprobed with an antibody to detect anti-AMPKα⊠, middle, and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH), bottom. (D) Left, 786-0 cells were treated with solvent control (DMSO), AICAR (500 µmol/L), fluvastatin (3 µmol/L), or the indicated combinations for 96 hours. Data are expressed as % apoptotic cells and represent means ± SE of four independent experiments. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with AICAR versus fluvastatin and AICAR showed a 2-tailed p value = 0.013. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with fluvastatin versus fluvastatin and AICAR treated cells showed a 2-tailed p value = 0.0005. Middle, 786-0 cells were treated with solvent control (DMSO), AICAR (500 µmol/L), simvastatin (3 µmol/L), or the indicated combinations for 96 hours. Data are expressed as % apoptotic cells and represent means ± SE of 3 independent experiments. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with AICAR versus simvastatin and AICAR showed a 2-tailed p value = 0.003. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with simvastatin versus simvastatin and AICAR showed a 2-tailed p value = 0.003. Right, ACHN cells were treated with solvent control (DMSO), AICAR (500 µmol/L), fluvastatin (3 µmol/L), or the indicated combinations for ninety-six hours. Data are expressed as % apoptotic cells and represent means ± SE of four independent experiments. Paired t-test analysis for the induction of apoptosis of ACHN cells treated with AICAR versus fluvastatin and AICAR showed a 2-tailed p value = 0.008. Paired t-test analysis for the induction of apoptosis of ACHN cells treated with fluvastatin versus fluvastatin and AICAR treated cells showed a 2-tailed p value = 0.003.

    Journal: Cancer Biology & Therapy

    Article Title: AMPK as a therapeutic target in renal cell carcinoma

    doi: 10.4161/cbt.10.11.13629

    Figure Lengend Snippet: AICAR-dependent growth suppression and apoptosis in renal cell carcinoma cells. (A) 786-0 cells were treated for 4 days with solvent control or with the indicated concentrations of AICAR. Cell proliferation was assessed by MTT. Data are expressed as % control-treated cells and represent means ± SE of three independent experiments. Paired t-test analysis for the growth of 786-0 cells treated with 250 µmol/L AICAR, 500 µmol/L AICAR and 1,000 µmol/L AICAR versus control treated cells showed a 2-tailed p values of 0.003, 0.0001 and 0.004, respectively. (B) Top, 786-0 cells were treated with solvent control or AICAR (2 mmol/L) for 48 hours, as indicated. Equal amounts of total cell lysates were resolved by SDS-PAGE and immunoblotted with an antibody against the phosphorylated/activated form of AMPKα at Thr 172. Middle, equal protein aliquots from the same experiment were resolved on a separate gel and probed with an anti-AMPKα antibody. Bottom, the top blot was subsequently reprobed with an antibody to detect anti-glyceradledhyde-3-phosphate dehydrogenase (anti-GAPDH). (C) Top, 786-0 cells were treated with solvent control or the indicated concentrations of AICAR for 48 hours. Equal amounts of total cell lysates were resolved by SDS-PAGE and immunoblotted with an antibody against the phosphorylated/activated form of AMPKα at Thr 172. Middle, bottom, the blot was subsequently stripped and reprobed with an antibody to detect anti-AMPKα⊠, middle, and anti-glyceraldehyde-3-phosphate dehydrogenase (anti-GAPDH), bottom. (D) Left, 786-0 cells were treated with solvent control (DMSO), AICAR (500 µmol/L), fluvastatin (3 µmol/L), or the indicated combinations for 96 hours. Data are expressed as % apoptotic cells and represent means ± SE of four independent experiments. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with AICAR versus fluvastatin and AICAR showed a 2-tailed p value = 0.013. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with fluvastatin versus fluvastatin and AICAR treated cells showed a 2-tailed p value = 0.0005. Middle, 786-0 cells were treated with solvent control (DMSO), AICAR (500 µmol/L), simvastatin (3 µmol/L), or the indicated combinations for 96 hours. Data are expressed as % apoptotic cells and represent means ± SE of 3 independent experiments. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with AICAR versus simvastatin and AICAR showed a 2-tailed p value = 0.003. Paired t-test analysis for the induction of apoptosis of 786-0 cells treated with simvastatin versus simvastatin and AICAR showed a 2-tailed p value = 0.003. Right, ACHN cells were treated with solvent control (DMSO), AICAR (500 µmol/L), fluvastatin (3 µmol/L), or the indicated combinations for ninety-six hours. Data are expressed as % apoptotic cells and represent means ± SE of four independent experiments. Paired t-test analysis for the induction of apoptosis of ACHN cells treated with AICAR versus fluvastatin and AICAR showed a 2-tailed p value = 0.008. Paired t-test analysis for the induction of apoptosis of ACHN cells treated with fluvastatin versus fluvastatin and AICAR treated cells showed a 2-tailed p value = 0.003.

    Article Snippet: An anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) monoclonal antibody was obtained from Millipore.

    Techniques: MTT Assay, SDS Page