anti gluk1  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92

    Structured Review

    Millipore anti gluk1
    The expression of KARs in the cultured cortical neurons from Fmr1 WT and Fmr1 KO mice. a , b , The abundance of <t>GluK1</t> or GluK2/3 in the homogenate of the cultured cortical neurons from Fmr1 WT mice and Fmr1 KO mice showed no change. c , d , Surface expression levels of GluK2/3 in insular cortex neurons obtained from Fmr1 WT and Fmr1 KO mice were detected by western blot analysis. The surface expression levels of GluK2/3 were not altered between Fmr1 WT and Fmr1 KO mice (n = 3 independent experiments). Actin was used as negative control for surface biotinylation
    Anti Gluk1, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 807 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gluk1/product/Millipore
    Average 92 stars, based on 807 article reviews
    Price from $9.99 to $1999.99
    anti gluk1 - by Bioz Stars, 2020-11
    92/100 stars

    Images

    1) Product Images from "Reduced synaptic function of Kainate receptors in the insular cortex of Fmr1 Knock-out mice"

    Article Title: Reduced synaptic function of Kainate receptors in the insular cortex of Fmr1 Knock-out mice

    Journal: Molecular Brain

    doi: 10.1186/s13041-018-0396-1

    The expression of KARs in the cultured cortical neurons from Fmr1 WT and Fmr1 KO mice. a , b , The abundance of GluK1 or GluK2/3 in the homogenate of the cultured cortical neurons from Fmr1 WT mice and Fmr1 KO mice showed no change. c , d , Surface expression levels of GluK2/3 in insular cortex neurons obtained from Fmr1 WT and Fmr1 KO mice were detected by western blot analysis. The surface expression levels of GluK2/3 were not altered between Fmr1 WT and Fmr1 KO mice (n = 3 independent experiments). Actin was used as negative control for surface biotinylation
    Figure Legend Snippet: The expression of KARs in the cultured cortical neurons from Fmr1 WT and Fmr1 KO mice. a , b , The abundance of GluK1 or GluK2/3 in the homogenate of the cultured cortical neurons from Fmr1 WT mice and Fmr1 KO mice showed no change. c , d , Surface expression levels of GluK2/3 in insular cortex neurons obtained from Fmr1 WT and Fmr1 KO mice were detected by western blot analysis. The surface expression levels of GluK2/3 were not altered between Fmr1 WT and Fmr1 KO mice (n = 3 independent experiments). Actin was used as negative control for surface biotinylation

    Techniques Used: Expressing, Cell Culture, Mouse Assay, Western Blot, Negative Control

    Distribution of KARs in the insular cortex from Fmr1 WT mice. Presynaptic marker synaptophysin, postsynaptic marker PSD95, NMDAR subunits GluN2B and GluN2A, KAR subunits GluK1 and GluK2/3 or synaptic protein Rab5 were analyzed by Western blot in the homogenates (H1, 10 μg), postnuclear supernatant (S1, 5 μg), nuclei and large debris pellet (P1, 10 μg), cytosomes (S2, 5 μg), crude synaptosomal membrane (P2, 5 μg), non-PSD (5 μg) or PSD (10 μg) fractions of the insular cortex in Fmr1 WT mice. This experiment was repeated three times
    Figure Legend Snippet: Distribution of KARs in the insular cortex from Fmr1 WT mice. Presynaptic marker synaptophysin, postsynaptic marker PSD95, NMDAR subunits GluN2B and GluN2A, KAR subunits GluK1 and GluK2/3 or synaptic protein Rab5 were analyzed by Western blot in the homogenates (H1, 10 μg), postnuclear supernatant (S1, 5 μg), nuclei and large debris pellet (P1, 10 μg), cytosomes (S2, 5 μg), crude synaptosomal membrane (P2, 5 μg), non-PSD (5 μg) or PSD (10 μg) fractions of the insular cortex in Fmr1 WT mice. This experiment was repeated three times

    Techniques Used: Mouse Assay, Marker, Western Blot

    The abundance of KARs in the synaptosome is decreased in Fmr1 KO mice. a , b , Total expression levels of GluK1 and GluK2/3 in the homogenates fraction of the insular cortex conducted from Fmr1 WT and Fmr1 KO mice were detected by Western blot. The expression levels of GluK1 and GluK2/3 in the homogenates were not altered between Fmr1 WT and Fmr1 KO mice ( n = 3 mice for each group). c , d , Expression levels of GluK1 and GluK2/3 in the synaptosome of the insular cortex obtained from Fmr1 WT and Fmr1 KO mice were detected by western blot anaylsis. The expression levels of GluK1 and GluK2/3 in the homogenates was significantly reduced in Fmr1 KO mice compare to Fmr1 WT mice ( n = 5 mice for each group). * P
    Figure Legend Snippet: The abundance of KARs in the synaptosome is decreased in Fmr1 KO mice. a , b , Total expression levels of GluK1 and GluK2/3 in the homogenates fraction of the insular cortex conducted from Fmr1 WT and Fmr1 KO mice were detected by Western blot. The expression levels of GluK1 and GluK2/3 in the homogenates were not altered between Fmr1 WT and Fmr1 KO mice ( n = 3 mice for each group). c , d , Expression levels of GluK1 and GluK2/3 in the synaptosome of the insular cortex obtained from Fmr1 WT and Fmr1 KO mice were detected by western blot anaylsis. The expression levels of GluK1 and GluK2/3 in the homogenates was significantly reduced in Fmr1 KO mice compare to Fmr1 WT mice ( n = 5 mice for each group). * P

    Techniques Used: Mouse Assay, Expressing, Western Blot

    2) Product Images from "Splice Variants of Perlucin from Haliotis laevigata Modulate the Crystallisation of CaCO3"

    Article Title: Splice Variants of Perlucin from Haliotis laevigata Modulate the Crystallisation of CaCO3

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0097126

    Amino acid sequences of Perlucin from H. laevigata . Perlucin splice variants (Perlucin-R0, Perlucin-R5, Perlucin-R8) are indicated as grey bars above the sequence. The following characteristics of the proteins are marked: Amino acid exchanges in Perlucin-R0 (M89I, V129D, R149L), signal peptide, C-type lectin domain [60] , repeat units (light blue bars). Peptides identified from 2D electrophoresis spots ( Figure 3 ) by MALDI-ToF MS (black bars) or ESI-MS (red bar). *Predicted glycosylation and phosphorylation (NetPhos [50] ) sites.
    Figure Legend Snippet: Amino acid sequences of Perlucin from H. laevigata . Perlucin splice variants (Perlucin-R0, Perlucin-R5, Perlucin-R8) are indicated as grey bars above the sequence. The following characteristics of the proteins are marked: Amino acid exchanges in Perlucin-R0 (M89I, V129D, R149L), signal peptide, C-type lectin domain [60] , repeat units (light blue bars). Peptides identified from 2D electrophoresis spots ( Figure 3 ) by MALDI-ToF MS (black bars) or ESI-MS (red bar). *Predicted glycosylation and phosphorylation (NetPhos [50] ) sites.

    Techniques Used: Sequencing, Two-Dimensional Gel Electrophoresis, Mass Spectrometry

    Electrophoretic analysis of recombinant and native Perlucin preparations. A) Western blot of cell culture supernatants derived from COS-7 cells ectopically over expressing the indicated Strep-tagged recombinant Perlucins, which were detected using a polyclonal anti-Strep-tag antibody as described in the Methods section. B) SDS-PAGE of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie brilliant blue showed one distinct band at approx. 25 kDa, one at 20 kDa, and one at approx. 15 kDa. C) 2D electrophoresis of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie Brilliant Blue. The indicated spots were cut out and analysed by MALDI-ToF MS as described in the Methods section. Spot 7 was used as control.
    Figure Legend Snippet: Electrophoretic analysis of recombinant and native Perlucin preparations. A) Western blot of cell culture supernatants derived from COS-7 cells ectopically over expressing the indicated Strep-tagged recombinant Perlucins, which were detected using a polyclonal anti-Strep-tag antibody as described in the Methods section. B) SDS-PAGE of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie brilliant blue showed one distinct band at approx. 25 kDa, one at 20 kDa, and one at approx. 15 kDa. C) 2D electrophoresis of native Perlucin purified from abalone shell of H. laevigata , stained with Coomassie Brilliant Blue. The indicated spots were cut out and analysed by MALDI-ToF MS as described in the Methods section. Spot 7 was used as control.

    Techniques Used: Recombinant, Western Blot, Cell Culture, Derivative Assay, Expressing, Strep-tag, SDS Page, Purification, Staining, Two-Dimensional Gel Electrophoresis, Mass Spectrometry

    3) Product Images from "Localization of tamoxifen in human breast cancer tumors by MALDI mass spectrometry imaging"

    Article Title: Localization of tamoxifen in human breast cancer tumors by MALDI mass spectrometry imaging

    Journal: Clinical and Translational Medicine

    doi: 10.1186/s40169-016-0090-9

    Ionization characteristics of tamoxifen (0.1 mg/mL in water) as measured with 3.5 mg/mL CHCA on a stainless steel MALDI target plate. a A full mass spectrum of tamoxifen obtained at 60,000 resolution using the Orbitrap mass analyzer and b a tandem mass spectrum of tamoxifen isolating the m/z 372.23 and CID fragmented in the linear ion trap mass analyzer
    Figure Legend Snippet: Ionization characteristics of tamoxifen (0.1 mg/mL in water) as measured with 3.5 mg/mL CHCA on a stainless steel MALDI target plate. a A full mass spectrum of tamoxifen obtained at 60,000 resolution using the Orbitrap mass analyzer and b a tandem mass spectrum of tamoxifen isolating the m/z 372.23 and CID fragmented in the linear ion trap mass analyzer

    Techniques Used:

    4) Product Images from "Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108"

    Article Title: Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2009.93

    Polysaccharide labeling with fluoresceinamine. CDAP-activated YCP reacted with fluoresceinamine for 18 h at room temperature. The mixtures were fractionated on a Sephacryl S-400 column, concentration (μg/mL) of fluoresceinamine (▴) and YCP (□) in each fraction were determined. n =3.
    Figure Legend Snippet: Polysaccharide labeling with fluoresceinamine. CDAP-activated YCP reacted with fluoresceinamine for 18 h at room temperature. The mixtures were fractionated on a Sephacryl S-400 column, concentration (μg/mL) of fluoresceinamine (▴) and YCP (□) in each fraction were determined. n =3.

    Techniques Used: Labeling, Concentration Assay

    5) Product Images from "Akt/PKB Controls the Activity-Dependent Bulk Endocytosis of Synaptic Vesicles"

    Article Title: Akt/PKB Controls the Activity-Dependent Bulk Endocytosis of Synaptic Vesicles

    Journal: Traffic (Copenhagen, Denmark)

    doi: 10.1111/j.1600-0854.2012.01365.x

    Effect of acute Akt inhibition on the uptake of large dextrans Cultures were incubated either in the absence or presence of Akt antagonists (Akti1/2, 500 n m , 10-NCP, 500 n m ) for 10 min. Cultures were then stimulated (80 Hz 10 seconds) in the presence of 50 μ m tetramethylrhodamine-dextran followed by immediate washout. A–C) Panels show representative fields of view of dextran loading in either control (Ctrl, A), or cultures treated with either Akti1/2 (B) or 10-NCP (C). Scale bar represents 20 µm. (D) Quantification of the extent of dextran uptake (puncta number per field) expressed as a percentage of control neurons ± SEM ( n = 3 independent experiments for all). One-way anova performed, all not significant.
    Figure Legend Snippet: Effect of acute Akt inhibition on the uptake of large dextrans Cultures were incubated either in the absence or presence of Akt antagonists (Akti1/2, 500 n m , 10-NCP, 500 n m ) for 10 min. Cultures were then stimulated (80 Hz 10 seconds) in the presence of 50 μ m tetramethylrhodamine-dextran followed by immediate washout. A–C) Panels show representative fields of view of dextran loading in either control (Ctrl, A), or cultures treated with either Akti1/2 (B) or 10-NCP (C). Scale bar represents 20 µm. (D) Quantification of the extent of dextran uptake (puncta number per field) expressed as a percentage of control neurons ± SEM ( n = 3 independent experiments for all). One-way anova performed, all not significant.

    Techniques Used: Inhibition, Incubation

    Akt phosphorylates GSK3 to retard dynamin I dephosphorylation during high intensity stimulation Cultures were incubated either in the absence or presence of Akt antagonists (Akti1/2–500 n m , 10-NCP–500 nM) for 10 min. Cultures were then either stimulated (80 Hz 10 seconds) or rested (basal) for 10 seconds in the absence and presence of antagonists and then immediately lysed. Representative blots display the phosphorylation of either (A) Akt Ser473, (B) GSK3 Ser9/21 or (C) dynamin I (DynI) Ser774 in the absence (− Drug) or presence (+ Drug) of either Akti1/2 or 10-NCP. The extent of phosphorylation/dephosphorylation of either Akt (A), GSK3 (B) or DynI (C) in the absence of inhibitor (Ctrl, clear bars), the presence of Akti1/2 (filled bars) or 10-NCP (hatched bars) is displayed. Data were corrected against protein levels (Syp) and expressed as the extent of stimulus-evoked phosphorylation over basal ± SEM ( n = 8 for PAkt control, n = 3 for PAkt Akti1/2, n = 3 for PAkt 10-NCP; n = 8 for PGSK3 control, n = 5 for PGSK3 Akti1/2, n = 5 for PGSK3 10-NCP; n = 17 for PDynI control, n = 13 for PDynI Akti1/2, n = 6 for PDynI 10-NCP). Student's t -test: *p
    Figure Legend Snippet: Akt phosphorylates GSK3 to retard dynamin I dephosphorylation during high intensity stimulation Cultures were incubated either in the absence or presence of Akt antagonists (Akti1/2–500 n m , 10-NCP–500 nM) for 10 min. Cultures were then either stimulated (80 Hz 10 seconds) or rested (basal) for 10 seconds in the absence and presence of antagonists and then immediately lysed. Representative blots display the phosphorylation of either (A) Akt Ser473, (B) GSK3 Ser9/21 or (C) dynamin I (DynI) Ser774 in the absence (− Drug) or presence (+ Drug) of either Akti1/2 or 10-NCP. The extent of phosphorylation/dephosphorylation of either Akt (A), GSK3 (B) or DynI (C) in the absence of inhibitor (Ctrl, clear bars), the presence of Akti1/2 (filled bars) or 10-NCP (hatched bars) is displayed. Data were corrected against protein levels (Syp) and expressed as the extent of stimulus-evoked phosphorylation over basal ± SEM ( n = 8 for PAkt control, n = 3 for PAkt Akti1/2, n = 3 for PAkt 10-NCP; n = 8 for PGSK3 control, n = 5 for PGSK3 Akti1/2, n = 5 for PGSK3 10-NCP; n = 17 for PDynI control, n = 13 for PDynI Akti1/2, n = 6 for PDynI 10-NCP). Student's t -test: *p

    Techniques Used: De-Phosphorylation Assay, Incubation

    6) Product Images from "Charge Derivatized Amino Acids Facilitate Model Studies on Protein Side-Chain Modifications by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry"

    Article Title: Charge Derivatized Amino Acids Facilitate Model Studies on Protein Side-Chain Modifications by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

    Journal: Rapid communications in mass spectrometry : RCM

    doi: 10.1002/rcm.4116

    MALDI-TOF mass spectra of reaction mixture of PLys and octenal (A) or heptenal (B) at various times.
    Figure Legend Snippet: MALDI-TOF mass spectra of reaction mixture of PLys and octenal (A) or heptenal (B) at various times.

    Techniques Used:

    7) Product Images from "Inhibition of tau fibrillization by oleocanthal via reaction with the amino groups of tau"

    Article Title: Inhibition of tau fibrillization by oleocanthal via reaction with the amino groups of tau

    Journal: Journal of neurochemistry

    doi: 10.1111/j.1471-4159.2009.06224.x

    Oleocanthal inhibits K18PL fibrillization in the presence of lysine. 0.158 μM to 1 mM lysine was added to the fibrillization of 20 μM K18PL. The reaction was incubated for 6 hrs at 37 °C and the fibrillization was measured by ThT
    Figure Legend Snippet: Oleocanthal inhibits K18PL fibrillization in the presence of lysine. 0.158 μM to 1 mM lysine was added to the fibrillization of 20 μM K18PL. The reaction was incubated for 6 hrs at 37 °C and the fibrillization was measured by ThT

    Techniques Used: Incubation

    Structure-activity relationship studies of oleocanthal and analogues. (a) Structures of oleocanthal analogues synthesized. (b, c, d) Effect of oleocanthal analogues on K18 fibrillization as monitored by ThT fluorescence assay. 10 μM compound is
    Figure Legend Snippet: Structure-activity relationship studies of oleocanthal and analogues. (a) Structures of oleocanthal analogues synthesized. (b, c, d) Effect of oleocanthal analogues on K18 fibrillization as monitored by ThT fluorescence assay. 10 μM compound is

    Techniques Used: Activity Assay, Synthesized, Fluorescence

    Oleocanthal inhibits K18 fibrillization. (a) ThT fluorescence assay of the filament assembly of K18 in the presence of 0-100 μM oleocanthal. AFUs , arbitrary fluorescence units. Error bars represent S.D.; n=3. *, P
    Figure Legend Snippet: Oleocanthal inhibits K18 fibrillization. (a) ThT fluorescence assay of the filament assembly of K18 in the presence of 0-100 μM oleocanthal. AFUs , arbitrary fluorescence units. Error bars represent S.D.; n=3. *, P

    Techniques Used: Fluorescence

    Oleocanthal inhibits T40 fibrillization. (a) ThT fluorescence assay of the filament assembly of T40 in the presence of 0-100 μM oleocanthal. AFUs , arbitrary fluorescence units. Error bars represent S.D.; n=3. *, P
    Figure Legend Snippet: Oleocanthal inhibits T40 fibrillization. (a) ThT fluorescence assay of the filament assembly of T40 in the presence of 0-100 μM oleocanthal. AFUs , arbitrary fluorescence units. Error bars represent S.D.; n=3. *, P

    Techniques Used: Fluorescence

    8) Product Images from "Identification of Clostridium spp. derived from a sheep and cattle slaughterhouse by matrix-assisted laser desorption and ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rDNA sequencing"

    Article Title: Identification of Clostridium spp. derived from a sheep and cattle slaughterhouse by matrix-assisted laser desorption and ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rDNA sequencing

    Journal: Journal of Food Science and Technology

    doi: 10.1007/s13197-018-3255-2

    Comparing direct transfer and extended direct transfer sample preparation methods for identification of Clostridium species by MALDI-TOF MS at different days of anaerobic incubation (1, 3, and 5). Each isolate was tested three times for each method of sample preparation at each day
    Figure Legend Snippet: Comparing direct transfer and extended direct transfer sample preparation methods for identification of Clostridium species by MALDI-TOF MS at different days of anaerobic incubation (1, 3, and 5). Each isolate was tested three times for each method of sample preparation at each day

    Techniques Used: Sample Prep, Mass Spectrometry, Incubation

    Overview of Bruker MALDI-TOF MS identification of Clostridium isolates at the species level. MALDI-TOF MS identified 94% isolates while 6% of isolates were not identified by MALDI-TOF MS. These isolates were identified via 16S rDNA as C. estertheticum (n = 8), C. frigidicarnis (n = 5), and C. gasigenes (n = 3) species
    Figure Legend Snippet: Overview of Bruker MALDI-TOF MS identification of Clostridium isolates at the species level. MALDI-TOF MS identified 94% isolates while 6% of isolates were not identified by MALDI-TOF MS. These isolates were identified via 16S rDNA as C. estertheticum (n = 8), C. frigidicarnis (n = 5), and C. gasigenes (n = 3) species

    Techniques Used: Mass Spectrometry

    9) Product Images from "UV laser-induced cross-linking in peptides"

    Article Title: UV laser-induced cross-linking in peptides

    Journal: Rapid communications in mass spectrometry : RCM

    doi: 10.1002/rcm.6610

    Positive-ion MALDI-TOF mass spectra of a mixture of xenopsin (M x ), interleukin (M i ) and angiotensin I (M a ) not irradiated (panel A) and irradiated for 10 sec (panel B).
    Figure Legend Snippet: Positive-ion MALDI-TOF mass spectra of a mixture of xenopsin (M x ), interleukin (M i ) and angiotensin I (M a ) not irradiated (panel A) and irradiated for 10 sec (panel B).

    Techniques Used: Irradiation, Size-exclusion Chromatography

    10) Product Images from "Blockade of mesolimbic dopamine D3 receptors inhibits stress-induced reinstatement of cocaine-seeking in rats"

    Article Title: Blockade of mesolimbic dopamine D3 receptors inhibits stress-induced reinstatement of cocaine-seeking in rats

    Journal: Psychopharmacology

    doi: 10.1007/s00213-004-1858-y

    Effects of local microinjections of the potent and the highly selective DA D 3 receptor antagonist SB-277011A into the nucleus accumbens ( NAcc ) ( A ) or dorsal striatum ( B ) on footshock-induced reinstatement of cocaine-seeking behavior. Statistical analyses revealed a significant stress-triggered reinstatement effect, and a significant protective effect against stress-triggered reinstatement by the highly selective DA D 3 receptor antagonist SB-277011A when microinjected bilaterally (1.5 μg/0.5 ml per side) into the NAcc but not into the dorsal striatum. See Results for details of the statistical analyses and findings
    Figure Legend Snippet: Effects of local microinjections of the potent and the highly selective DA D 3 receptor antagonist SB-277011A into the nucleus accumbens ( NAcc ) ( A ) or dorsal striatum ( B ) on footshock-induced reinstatement of cocaine-seeking behavior. Statistical analyses revealed a significant stress-triggered reinstatement effect, and a significant protective effect against stress-triggered reinstatement by the highly selective DA D 3 receptor antagonist SB-277011A when microinjected bilaterally (1.5 μg/0.5 ml per side) into the NAcc but not into the dorsal striatum. See Results for details of the statistical analyses and findings

    Techniques Used:

    Effects of systemic administration of the potent and highly selective DA D 3 receptor antagonist SB-277011A on footshock-induced reinstatement of cocaine-seeking behavior. A Mean responding on the active lever during the last session of cocaine self-administration ( SA ), during the last session of extinction prior to testing with either vehicle or SB-277011A, and during the test session for footshock-induced reinstatement in the presence of vehicle or SB-277011A. Statistical analyses revealed a significant stress-triggered reinstatement effect, and a significant dose-dependent protective effect against stress-triggered reinstatement by the highly selective DA D 3 receptor antagonist SB-277011A. B Mean responding on the inactive lever. See Results for details of the statistical analyses and findings
    Figure Legend Snippet: Effects of systemic administration of the potent and highly selective DA D 3 receptor antagonist SB-277011A on footshock-induced reinstatement of cocaine-seeking behavior. A Mean responding on the active lever during the last session of cocaine self-administration ( SA ), during the last session of extinction prior to testing with either vehicle or SB-277011A, and during the test session for footshock-induced reinstatement in the presence of vehicle or SB-277011A. Statistical analyses revealed a significant stress-triggered reinstatement effect, and a significant dose-dependent protective effect against stress-triggered reinstatement by the highly selective DA D 3 receptor antagonist SB-277011A. B Mean responding on the inactive lever. See Results for details of the statistical analyses and findings

    Techniques Used:

    Diagrammatic sequence of experimental phases. Each animal received two reinstatement tests after extinction, once with vehicle and once with one dose of the potent and highly selective DA D 3 receptor antagonist SB-277011A in a counterbalanced manner
    Figure Legend Snippet: Diagrammatic sequence of experimental phases. Each animal received two reinstatement tests after extinction, once with vehicle and once with one dose of the potent and highly selective DA D 3 receptor antagonist SB-277011A in a counterbalanced manner

    Techniques Used: Sequencing

    11) Product Images from "Immunocontraceptive target repertoire defined by systematic identification of sperm membrane alloantigens in a single species"

    Article Title: Immunocontraceptive target repertoire defined by systematic identification of sperm membrane alloantigens in a single species

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0190891

    Fertility of gilts immunized with sperm membranes or lipid rafts. A) Detection of SMA with anti-TWM alloantisera from four different immunized gilts. Blots of TWM (triple-washed membranes, 50 μg protein/gel) resolved by 2-D gel electrophoresis were each probed with individual alloantisera (1/5000 dilution, sera collected 4 wk after boost), and immunoreactivity visualized with identical exposure times. Note the variability of the immune response evident as the high immunoreactivity of sera from two gilts (“High Ab,” #466 474), and lower immunoreactivity of two others (“Low Ab,” #465 473). B) Fertility of immunized gilts (TWM n = 4; lipid rafts n = 2) compared to gilts injected with adjuvant only (
    Figure Legend Snippet: Fertility of gilts immunized with sperm membranes or lipid rafts. A) Detection of SMA with anti-TWM alloantisera from four different immunized gilts. Blots of TWM (triple-washed membranes, 50 μg protein/gel) resolved by 2-D gel electrophoresis were each probed with individual alloantisera (1/5000 dilution, sera collected 4 wk after boost), and immunoreactivity visualized with identical exposure times. Note the variability of the immune response evident as the high immunoreactivity of sera from two gilts (“High Ab,” #466 474), and lower immunoreactivity of two others (“Low Ab,” #465 473). B) Fertility of immunized gilts (TWM n = 4; lipid rafts n = 2) compared to gilts injected with adjuvant only ("FCA", n = 4). Gilts were bred on second estrus after collection of alloantisera used for the blots in Panel A. Data are expressed as mean litter size ± SD, with results for TWM-immunized high responder ("High Ab" blots in panel A) and low responder animals shown separately. For reference, shown also is our swine farm’s average litter size for non-immunized sows ("None") over a preceding 7-month period (2528 offspring/247 litters = 10.3 ± 0.6). C) Effect of alloimmunization on fertility. We compared litter sizes of high responder gilts immunized with TWM or lipid rafts to litter sizes of gilts injected with Freund's Complete Adjuvant only ("FCA"). Shown are mean litter sizes and standard error of the means with n = 4 in each group. High immunity to SMA or SLRA produced a 3.8-fold decrease in litter size (from 10.5 to 3.3 piglets/litter, p = 0.0119).

    Techniques Used: Nucleic Acid Electrophoresis, Injection, Produced

    12) Product Images from "Prostaglandin F2α-F-prostanoid receptor regulates CXCL8 expression in endometrial adenocarcinoma cells via the calcium-calcineurin-NFAT pathway"

    Article Title: Prostaglandin F2α-F-prostanoid receptor regulates CXCL8 expression in endometrial adenocarcinoma cells via the calcium-calcineurin-NFAT pathway

    Journal: Biochimica et Biophysica Acta

    doi: 10.1016/j.bbamcr.2009.09.018

    PGF 2α -FP receptor-mediated CXCL8 is regulated via the calcium–calcineurin–NFAT pathway. (A) CXCL8 mRNA expression and (B) protein secretion in Ishikawa FPS cells treated for 8 h with vehicle, 100 nM PGF 2α , 100 nM PGF 2α in the absence/presence of AL8810 (50 μM), YM254890 (1 μM), U73122 (10 μM), 4C3MQ (1 μM), RO-318220 (1 μM ), EGTA (1.5 mM), Inca-6 (40 μM) or CsA (1 μM) as determined by quantitative RT-PCR analysis and ELISA respectively. (C) CXCL8 luciferase activity in Ishikawa FPS cells transiently transfected with the full length (− 1400/+ 44) or sequential 5′ deletions (− 162/+ 44, − 132/+ 44, − 66/+ 44, − 54/+ 44) of the CXCL8 promoter or the − 162/+ 44 construct containing site-directed mutations in the NFAT or AP1 binding sites. Cells were treated with vehicle or 100 nM PGF 2α for 8 h and expressed as fold above the − 54/+ 44 construct which contains only the TATA box. (D) FPS cells were transiently transfected with the full length − 1400/+ 44 CXCL8 promoter and treated with vehicle or 100 nM PGF 2α for 8 h in the absence/presence of AL8810, YM254890, EGTA, CsA, Inca-6 or SN-50. CXCL8 promoter activity was determined by luciferase reporter assay. (E) FPS cells were transiently transfected with pAP1-luc, pNFAT-luc or pNF NFκB -luc reporter cDNA constructs and treated for 2, 4, 8 or 24 h with vehicle or 100 nM PGF 2α . (F) CXCL8 luciferase activity in FPS cells transfected with the − 162/+ 44 CXCL8 luciferase reporter plasmid. Cells were treated with vehicle or 100 nM PGF 2α for 8 h in absence/presence of Inca-6, SN-50 or transretinoic acid (RA). (b is significantly different from a; P
    Figure Legend Snippet: PGF 2α -FP receptor-mediated CXCL8 is regulated via the calcium–calcineurin–NFAT pathway. (A) CXCL8 mRNA expression and (B) protein secretion in Ishikawa FPS cells treated for 8 h with vehicle, 100 nM PGF 2α , 100 nM PGF 2α in the absence/presence of AL8810 (50 μM), YM254890 (1 μM), U73122 (10 μM), 4C3MQ (1 μM), RO-318220 (1 μM ), EGTA (1.5 mM), Inca-6 (40 μM) or CsA (1 μM) as determined by quantitative RT-PCR analysis and ELISA respectively. (C) CXCL8 luciferase activity in Ishikawa FPS cells transiently transfected with the full length (− 1400/+ 44) or sequential 5′ deletions (− 162/+ 44, − 132/+ 44, − 66/+ 44, − 54/+ 44) of the CXCL8 promoter or the − 162/+ 44 construct containing site-directed mutations in the NFAT or AP1 binding sites. Cells were treated with vehicle or 100 nM PGF 2α for 8 h and expressed as fold above the − 54/+ 44 construct which contains only the TATA box. (D) FPS cells were transiently transfected with the full length − 1400/+ 44 CXCL8 promoter and treated with vehicle or 100 nM PGF 2α for 8 h in the absence/presence of AL8810, YM254890, EGTA, CsA, Inca-6 or SN-50. CXCL8 promoter activity was determined by luciferase reporter assay. (E) FPS cells were transiently transfected with pAP1-luc, pNFAT-luc or pNF NFκB -luc reporter cDNA constructs and treated for 2, 4, 8 or 24 h with vehicle or 100 nM PGF 2α . (F) CXCL8 luciferase activity in FPS cells transfected with the − 162/+ 44 CXCL8 luciferase reporter plasmid. Cells were treated with vehicle or 100 nM PGF 2α for 8 h in absence/presence of Inca-6, SN-50 or transretinoic acid (RA). (b is significantly different from a; P

    Techniques Used: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Luciferase, Activity Assay, Transfection, Construct, Binding Assay, Reporter Assay, Plasmid Preparation

    The AP1 protein c-Jun activates the AP1 and NFAT cis-acting enhancer elements to transactivate the CXCL8 promoter. FPS cells were transiently transfected with pNFAT-luc (A) or pAP1-luc (B) and treated for 8 h with vehicle, 100 nM PGF 2α , 100 nM PGF 2α in the absence/presence of AL8810, YM254890, U73122, 4C3MQ, RO-318220, EGTA, Inca-6 or CsA and NFAT cis-acting enhancer DNA activation was measured by luciferase reporter assay. Ishikawa FPS cells were transiently transfected with the pAP1-luc (C) or pNFAT-luc (D) reporter plasmid and either incubated with transretinoic acid or co-transfected with an empty vector or dn c-Jun cDNA construct. Ishikawa FPS cells were transfected with the − 162/+ 44 or − 1400/+ 44 (E) CXCL8 luciferase reporter cDNA and co-transfected with empty vector or dn c-Jun cDNA. Cells were treated with vehicle or 100 nM PGF 2α for 8 h. (b is significantly different from a and c is significantly different from a and b; P
    Figure Legend Snippet: The AP1 protein c-Jun activates the AP1 and NFAT cis-acting enhancer elements to transactivate the CXCL8 promoter. FPS cells were transiently transfected with pNFAT-luc (A) or pAP1-luc (B) and treated for 8 h with vehicle, 100 nM PGF 2α , 100 nM PGF 2α in the absence/presence of AL8810, YM254890, U73122, 4C3MQ, RO-318220, EGTA, Inca-6 or CsA and NFAT cis-acting enhancer DNA activation was measured by luciferase reporter assay. Ishikawa FPS cells were transiently transfected with the pAP1-luc (C) or pNFAT-luc (D) reporter plasmid and either incubated with transretinoic acid or co-transfected with an empty vector or dn c-Jun cDNA construct. Ishikawa FPS cells were transfected with the − 162/+ 44 or − 1400/+ 44 (E) CXCL8 luciferase reporter cDNA and co-transfected with empty vector or dn c-Jun cDNA. Cells were treated with vehicle or 100 nM PGF 2α for 8 h. (b is significantly different from a and c is significantly different from a and b; P

    Techniques Used: Transfection, Activation Assay, Luciferase, Reporter Assay, Plasmid Preparation, Incubation, Construct

    13) Product Images from "Characterization of Bacillus subtilis Colony Biofilms via Mass Spectrometry and Fluorescence Imaging"

    Article Title: Characterization of Bacillus subtilis Colony Biofilms via Mass Spectrometry and Fluorescence Imaging

    Journal: Journal of Proteome Research

    doi: 10.1021/acs.jproteome.6b00127

    Fluorescence imaging and MALDI MSI of B. subtilis biofilms of NCIB3610 and its mutants integrated with the P yqxM -CFP reporter. CFP images were acquired using a fluorescence stereoscope before MALDI analysis. Each individual column represents (left to right): m / z 1030 (surfactin-C13, [M + Na] + ); m / z 1044 (surfactin-C14, [M + Na] + ); m / z 1058 (surfactin-C15, [M + Na] + ); m / z 1485 (plipastatin-C16-Ala, [M + Na] + ); m / z 1499 (plipastatin-C17-Ala, [M + Na] + ); m / z 1513 (plipastatin-C16-Val, [M + Na] + ); m / z 1527 (plipastatin-C17-Val, [M + Na] + ); m / z 2782 (SKF, [M + H] + ), m / z 3422 (subtilosin, [M + Na] + ), m / z 4334 (SDP, [M + Na] + ), and an overlay of ion images (green: SKF; bright pink: SDP). The ion intensity is reflected by the intensity of colors. Each column of ions is displayed using the same intensity scale, optimized per each metabolite and normalized to the TIC. Scale bar = 2 mm for fluorescence images and 5 mm for optical and ion images.
    Figure Legend Snippet: Fluorescence imaging and MALDI MSI of B. subtilis biofilms of NCIB3610 and its mutants integrated with the P yqxM -CFP reporter. CFP images were acquired using a fluorescence stereoscope before MALDI analysis. Each individual column represents (left to right): m / z 1030 (surfactin-C13, [M + Na] + ); m / z 1044 (surfactin-C14, [M + Na] + ); m / z 1058 (surfactin-C15, [M + Na] + ); m / z 1485 (plipastatin-C16-Ala, [M + Na] + ); m / z 1499 (plipastatin-C17-Ala, [M + Na] + ); m / z 1513 (plipastatin-C16-Val, [M + Na] + ); m / z 1527 (plipastatin-C17-Val, [M + Na] + ); m / z 2782 (SKF, [M + H] + ), m / z 3422 (subtilosin, [M + Na] + ), m / z 4334 (SDP, [M + Na] + ), and an overlay of ion images (green: SKF; bright pink: SDP). The ion intensity is reflected by the intensity of colors. Each column of ions is displayed using the same intensity scale, optimized per each metabolite and normalized to the TIC. Scale bar = 2 mm for fluorescence images and 5 mm for optical and ion images.

    Techniques Used: Fluorescence, Imaging

    14) Product Images from "Proteolytic Cleavage of Human Chromogranin A Containing Naturally Occurring Catestatin Variants: Differential Processing at Catestatin Region by Plasmin"

    Article Title: Proteolytic Cleavage of Human Chromogranin A Containing Naturally Occurring Catestatin Variants: Differential Processing at Catestatin Region by Plasmin

    Journal:

    doi: 10.1210/en.2007-0838

    MALDI-TOF of recombinant WT and mutant human CgA proteins digested with plasmin. Purified protein (10 μM/reaction) was incubated for 15 min with: 2 μM plasmin (A, left panel ) and 1 μM plasmin (A, right panel ); and 0.2 μ
    Figure Legend Snippet: MALDI-TOF of recombinant WT and mutant human CgA proteins digested with plasmin. Purified protein (10 μM/reaction) was incubated for 15 min with: 2 μM plasmin (A, left panel ) and 1 μM plasmin (A, right panel ); and 0.2 μ

    Techniques Used: Recombinant, Mutagenesis, Purification, Incubation

    MALDI-TOF of synthetic catestatin isoform peptides subjected to digestion with plasmin. A, Synthetic peptides (human CgA 360–380 ), wild type (ARAYGFRGPGP 370 QLRRGWRPSS; m/z = 2372.2), or Pro370Leu (ARAYGFRGPGL 370 QLRRGWRPSS; m/z =
    Figure Legend Snippet: MALDI-TOF of synthetic catestatin isoform peptides subjected to digestion with plasmin. A, Synthetic peptides (human CgA 360–380 ), wild type (ARAYGFRGPGP 370 QLRRGWRPSS; m/z = 2372.2), or Pro370Leu (ARAYGFRGPGL 370 QLRRGWRPSS; m/z =

    Techniques Used:

    15) Product Images from "Nuclear Magnetic Resonance seq (NMRseq): A New Approach to Peptide Sequence Tags"

    Article Title: Nuclear Magnetic Resonance seq (NMRseq): A New Approach to Peptide Sequence Tags

    Journal: Toxins

    doi: 10.3390/toxins10110437

    Characterisation of Conus geographus venom peptides. ( A ) Semi-preparative reversed-phase high performance liquid chromatography (RP-HPLC) chromatogram of crude Conus geographus venom with the peaks corresponding to the two peptides conopressin G (Con-G; mass 1033.4459 Da) and G11.1 (mass 3202.0503 Da), respectively, highlighted in red (C 18 Phenomenex Jupiter 250 × 10 mm, 10 µm, 300 Å; 3 mL/min flow rate; 0–60% Solvent B in 120 min (Solvent A: 0.05% trifluoroacetic acid (TFA); Solvent B: 90% acetonitrile, 0.045% TFA), 60–90% Solvent B in 5 min, 90% Solvent B for 10 min, 90–0% Solvent B in 5 min; UV absorbance at 214 nm and 280 nm); ( B ) SCIEX TOF/TOF™ 5800 matrix-assisted laser desorption ionization (MALDI) MS spectrum of the conopressin G fraction (mass 1033.4459 Da) using α–cyano-4-hydroxycinnamic acid (CHCA) matrix with the isotope distribution shown expanded inset; ( C ) SCIEX TOF/TOF™ 5800 MALDI MS spectrum of the G11.1 fraction (mass 3202.0503 Da) using CHCA matrix with the isotope distribution shown expanded inset.
    Figure Legend Snippet: Characterisation of Conus geographus venom peptides. ( A ) Semi-preparative reversed-phase high performance liquid chromatography (RP-HPLC) chromatogram of crude Conus geographus venom with the peaks corresponding to the two peptides conopressin G (Con-G; mass 1033.4459 Da) and G11.1 (mass 3202.0503 Da), respectively, highlighted in red (C 18 Phenomenex Jupiter 250 × 10 mm, 10 µm, 300 Å; 3 mL/min flow rate; 0–60% Solvent B in 120 min (Solvent A: 0.05% trifluoroacetic acid (TFA); Solvent B: 90% acetonitrile, 0.045% TFA), 60–90% Solvent B in 5 min, 90% Solvent B for 10 min, 90–0% Solvent B in 5 min; UV absorbance at 214 nm and 280 nm); ( B ) SCIEX TOF/TOF™ 5800 matrix-assisted laser desorption ionization (MALDI) MS spectrum of the conopressin G fraction (mass 1033.4459 Da) using α–cyano-4-hydroxycinnamic acid (CHCA) matrix with the isotope distribution shown expanded inset; ( C ) SCIEX TOF/TOF™ 5800 MALDI MS spectrum of the G11.1 fraction (mass 3202.0503 Da) using CHCA matrix with the isotope distribution shown expanded inset.

    Techniques Used: High Performance Liquid Chromatography, Flow Cytometry, Mass Spectrometry

    16) Product Images from "Lactate, a putative survival factor for myeloma cells, is incorporated by myeloma cells through monocarboxylate transporters 1"

    Article Title: Lactate, a putative survival factor for myeloma cells, is incorporated by myeloma cells through monocarboxylate transporters 1

    Journal: Experimental Hematology & Oncology

    doi: 10.1186/s40164-015-0008-z

    Schematic model of lactate kinetics between myeloma cells and bone marrow stromal cells. Lactate is produced by both myeloma cells and bone marrow stromal cells, suggesting that it acts as an autocrine and paracrine energy source. MCT1 may play an important role as a gate for incoming lactate. Targeting of MCT1 may result in decreased incorporation of lactate, resulting in cell death. PDH; pyruvate dehydrogenase, PDHK; pyruvate dehydrogenase kinase, MCT1; monocarboxylate transporter1, DCA; Dichloroacetate CHC; α-cyano-4-hydroxycinnamic acid.
    Figure Legend Snippet: Schematic model of lactate kinetics between myeloma cells and bone marrow stromal cells. Lactate is produced by both myeloma cells and bone marrow stromal cells, suggesting that it acts as an autocrine and paracrine energy source. MCT1 may play an important role as a gate for incoming lactate. Targeting of MCT1 may result in decreased incorporation of lactate, resulting in cell death. PDH; pyruvate dehydrogenase, PDHK; pyruvate dehydrogenase kinase, MCT1; monocarboxylate transporter1, DCA; Dichloroacetate CHC; α-cyano-4-hydroxycinnamic acid.

    Techniques Used: Produced

    17) Product Images from "The adipocyte differentiation protein APMAP is an endogenous suppressor of Aβ production in the brain"

    Article Title: The adipocyte differentiation protein APMAP is an endogenous suppressor of Aβ production in the brain

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddu449

    APMAP controls the levels and the stability of APP-CTFs. ( A ) siRNA knockdown of APMAP in HeLa cells quantitatively and qualitatively impacts APP-CTFs. Reduced APMAP expression correlates with increased APP-CTFs and the formation of three forms of APP-CTFα (CTFα1, α2 and α3), as revealed on a Tris-Tricine urea gel (left panel). This phenotype is further amplified in the presence of 1 µ m GSI DAPT (right panel). Biological triplicates are shown. ( B ) MALDI-TOF mass spectrometric analysis of APP-CTFα1, α2 and α3 immunoprecipitated from DAPT-treated cells with siRNA-reduced APMAP expression. ( C ) Peptide sequences of N-terminal truncated APP-CTFα1 (C80), -CTFα2 (C74) and -CTFα3 (C71).
    Figure Legend Snippet: APMAP controls the levels and the stability of APP-CTFs. ( A ) siRNA knockdown of APMAP in HeLa cells quantitatively and qualitatively impacts APP-CTFs. Reduced APMAP expression correlates with increased APP-CTFs and the formation of three forms of APP-CTFα (CTFα1, α2 and α3), as revealed on a Tris-Tricine urea gel (left panel). This phenotype is further amplified in the presence of 1 µ m GSI DAPT (right panel). Biological triplicates are shown. ( B ) MALDI-TOF mass spectrometric analysis of APP-CTFα1, α2 and α3 immunoprecipitated from DAPT-treated cells with siRNA-reduced APMAP expression. ( C ) Peptide sequences of N-terminal truncated APP-CTFα1 (C80), -CTFα2 (C74) and -CTFα3 (C71).

    Techniques Used: Expressing, Amplification, Immunoprecipitation

    18) Product Images from "Substantia Nigra D1 Receptors and Stimulation of Striatal Cholinergic Interneurons by Dopamine: A Proposed Circuit Mechanism"

    Article Title: Substantia Nigra D1 Receptors and Stimulation of Striatal Cholinergic Interneurons by Dopamine: A Proposed Circuit Mechanism

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.17-21-08498.1997

    Left ). •, Sites at which injection of the D 1  agonist SKF 38393 (0.5 μg/0.5 μl) was effective in increasing striatal ACh efflux. ▴, Injection sites of the aCSF vehicle (0.5 μl). ♦, Site at which injection of SKF 38393 (0.5 μg/0.5 μl) had no significant effect on striatal ACh efflux.  Right , Effect of intranigral injection of the D 1  agonist SKF 38393 (0.5 μg/0.5 μl; circles ) or the aCSF vehicle (0.5 μl; triangles ) on the output of striatal ACh. Injection of SKF 38393 or aCSF took place between minutes 5 and 10 of the 15 min dialysis sample ( arrow ). Data are femtomoles of ACh/20 μl sample of dialysate (mean ± SEM). * p
    Figure Legend Snippet: Left ). •, Sites at which injection of the D 1 agonist SKF 38393 (0.5 μg/0.5 μl) was effective in increasing striatal ACh efflux. ▴, Injection sites of the aCSF vehicle (0.5 μl). ♦, Site at which injection of SKF 38393 (0.5 μg/0.5 μl) had no significant effect on striatal ACh efflux. Right , Effect of intranigral injection of the D 1 agonist SKF 38393 (0.5 μg/0.5 μl; circles ) or the aCSF vehicle (0.5 μl; triangles ) on the output of striatal ACh. Injection of SKF 38393 or aCSF took place between minutes 5 and 10 of the 15 min dialysis sample ( arrow ). Data are femtomoles of ACh/20 μl sample of dialysate (mean ± SEM). * p

    Techniques Used: Injection

    19) Product Images from "Benchtop Micromolding of Polystyrene by Soft Lithography"

    Article Title: Benchtop Micromolding of Polystyrene by Soft Lithography

    Journal: Lab on a chip

    doi: 10.1039/c1lc20281b

    PS microfluidic chip. (A) Schematic of thermal bonding to form an enclosed microfluidic channel. (B) A prototype of a PS microfluidic chip. Rhodamine B (10 mM) was used for visualization of the microchannel (200 µm width, 50 µm depth).
    Figure Legend Snippet: PS microfluidic chip. (A) Schematic of thermal bonding to form an enclosed microfluidic channel. (B) A prototype of a PS microfluidic chip. Rhodamine B (10 mM) was used for visualization of the microchannel (200 µm width, 50 µm depth).

    Techniques Used: Chromatin Immunoprecipitation

    Rhodamine B adsorption/absorption tests. Fluorescence images of PS (A and B) and PDMS (C and D) devices are shown. The channels on these devices are filled with 100 µM rhodamine B in water for 15 min, and then rinsed with water. Fluorescent profiles
    Figure Legend Snippet: Rhodamine B adsorption/absorption tests. Fluorescence images of PS (A and B) and PDMS (C and D) devices are shown. The channels on these devices are filled with 100 µM rhodamine B in water for 15 min, and then rinsed with water. Fluorescent profiles

    Techniques Used: Adsorption, Fluorescence

    20) Product Images from "Evidence for the Effectiveness of Remdesivir (GS-5734), a Nucleoside-Analog Antiviral Drug in the Inhibition of IK(M) or IK(DR) and in the Stimulation of IMEP"

    Article Title: Evidence for the Effectiveness of Remdesivir (GS-5734), a Nucleoside-Analog Antiviral Drug in the Inhibition of IK(M) or IK(DR) and in the Stimulation of IMEP

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2020.01091

    The stimulatory effect of RDV on membrane electroporation-induced current ( I MEP ) identified in GH 3 cells. In this separate set of experiments, we bathed cells in Ca 2+ -free, Tyrode’s solution and the recording pipette was filled with K + -containing solution. (A) Representative I MEP traces obtained in the control ( 1 ) and during cell exposure to 3 μM RDV ( 2 ) or 10 μM RDV ( 3 ). The voltage-clamp protocol applied is denoted in the upper part, arrowhead is the zero current level and the calibration mark at the right lower part applies all current traces. Noticeably, the addition of RDV causes a measurable increase in the amplitude of I MEP elicited by large membrane hyperpolarization from −80 to −200 mV with a duration of 300 ms. (B) Concentration-dependent stimulation of I MEP produced by RDV in GH 3 cells (mean ± SEM; n=8 for each point). Current amplitude was measured at the end of hyperpolarizing pulse from −80 to −200 mV with a duration of 300 ms, and the vertical dashed line is placed at the IC 50 value required for RDV-stimulated I MEP . (C) Summary bar graph showing effect of RDV, RDV plus ivabradine (IVA) and RDV plus LaCl 3 on I MEP amplitude (mean ± SEM; n=8 for each bar). Current amplitude was taken at the end of hyperpolarizing voltage pulse from −80 to −200 mV with a duration of 300 ms. * Significantly different from control ( P
    Figure Legend Snippet: The stimulatory effect of RDV on membrane electroporation-induced current ( I MEP ) identified in GH 3 cells. In this separate set of experiments, we bathed cells in Ca 2+ -free, Tyrode’s solution and the recording pipette was filled with K + -containing solution. (A) Representative I MEP traces obtained in the control ( 1 ) and during cell exposure to 3 μM RDV ( 2 ) or 10 μM RDV ( 3 ). The voltage-clamp protocol applied is denoted in the upper part, arrowhead is the zero current level and the calibration mark at the right lower part applies all current traces. Noticeably, the addition of RDV causes a measurable increase in the amplitude of I MEP elicited by large membrane hyperpolarization from −80 to −200 mV with a duration of 300 ms. (B) Concentration-dependent stimulation of I MEP produced by RDV in GH 3 cells (mean ± SEM; n=8 for each point). Current amplitude was measured at the end of hyperpolarizing pulse from −80 to −200 mV with a duration of 300 ms, and the vertical dashed line is placed at the IC 50 value required for RDV-stimulated I MEP . (C) Summary bar graph showing effect of RDV, RDV plus ivabradine (IVA) and RDV plus LaCl 3 on I MEP amplitude (mean ± SEM; n=8 for each bar). Current amplitude was taken at the end of hyperpolarizing voltage pulse from −80 to −200 mV with a duration of 300 ms. * Significantly different from control ( P

    Techniques Used: Electroporation, Transferring, Concentration Assay, Produced

    21) Product Images from "Synchronization through nonreciprocal connections in a hybrid hippocampus microcircuit"

    Article Title: Synchronization through nonreciprocal connections in a hybrid hippocampus microcircuit

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2013.00120

    NMDA block alone is sufficient to disrupt synchrony in Circuit 5. (A) Top , Example traces from the two PCs in (Circuit 5) in the presence of both dAP5 (50 μM) and CNQX (10 μM). Bottom , The same cell recordings as in “ A ” with the further application of PTX (10 μM). Gray box shows a 100 ms interval around spikes form one of the PCs (dotted line). Scale bars: 10 mV, 100 ms. (B) Top , Mean Cross-correlograms of spike trains from circuit 5 without synaptic blockers; middle , in the presence of dAP5 and CNQX; bottom , in the presence of dAP5, CNQX and PTX. (C) Box plots of mean SI ( * p = 0.04).
    Figure Legend Snippet: NMDA block alone is sufficient to disrupt synchrony in Circuit 5. (A) Top , Example traces from the two PCs in (Circuit 5) in the presence of both dAP5 (50 μM) and CNQX (10 μM). Bottom , The same cell recordings as in “ A ” with the further application of PTX (10 μM). Gray box shows a 100 ms interval around spikes form one of the PCs (dotted line). Scale bars: 10 mV, 100 ms. (B) Top , Mean Cross-correlograms of spike trains from circuit 5 without synaptic blockers; middle , in the presence of dAP5 and CNQX; bottom , in the presence of dAP5, CNQX and PTX. (C) Box plots of mean SI ( * p = 0.04).

    Techniques Used: Blocking Assay, Mass Spectrometry

    Native NMDA receptors are important for pyramidal cell synchronization in a hybrid CA1 neuronal circuit. (A) left , Example traces from (Circuit 5) after bath application of the NMDA receptor blocker dAP5 (50 μM); middle , Mean cross-correlograms of (Circuit 5) before and after dAP5; right , Summary of SI for each pair of PCs before and after dAP5 ( * p = 0.03, n = 9). (B) left , Example traces from circuit 5 before and after application of the AMPA receptor blocker CNQX (10 μM) application; middle , Mean cross-correlation plots of Circuit 5 before and after CNQX; right , Summary of SI for each pair of PCs before and after CNQX ( p = 0.66, n = 6). (C) left , Example traces from Circuit 5 before and after application of the GABA receptor blocker PTX (10 μM); middle , Mean cross-correlation plots of circuit 5 before and after PTX; right , Summary of SI for each pair of PCs before and after PTX (10 μM) ( p = 0.43, n = 8). Gray box shows a time interval of 100 ms around one of the PCs firing (dotted line). Scale bars 10 mV, 100 ms.
    Figure Legend Snippet: Native NMDA receptors are important for pyramidal cell synchronization in a hybrid CA1 neuronal circuit. (A) left , Example traces from (Circuit 5) after bath application of the NMDA receptor blocker dAP5 (50 μM); middle , Mean cross-correlograms of (Circuit 5) before and after dAP5; right , Summary of SI for each pair of PCs before and after dAP5 ( * p = 0.03, n = 9). (B) left , Example traces from circuit 5 before and after application of the AMPA receptor blocker CNQX (10 μM) application; middle , Mean cross-correlation plots of Circuit 5 before and after CNQX; right , Summary of SI for each pair of PCs before and after CNQX ( p = 0.66, n = 6). (C) left , Example traces from Circuit 5 before and after application of the GABA receptor blocker PTX (10 μM); middle , Mean cross-correlation plots of circuit 5 before and after PTX; right , Summary of SI for each pair of PCs before and after PTX (10 μM) ( p = 0.43, n = 8). Gray box shows a time interval of 100 ms around one of the PCs firing (dotted line). Scale bars 10 mV, 100 ms.

    Techniques Used: Mass Spectrometry

    22) Product Images from "Agrp neuron activity is required for alcohol-induced overeating"

    Article Title: Agrp neuron activity is required for alcohol-induced overeating

    Journal: Nature Communications

    doi: 10.1038/ncomms14014

    Effects of EtOH on membrane electrical properties of Agrp neurons. ( a ) Effect of 50 mM EtOH on spike rate in cell-attached recordings. Representative example of six cells. Firing before EtOH=1.85±0.35 Hz, after=3.72±0.52 Hz, paired t -test: t =4.613, d.f.=5, P =0.0058. ( b ) Effect of 50 mM EtOH on membrane potential, under synaptic blockade with of 10 μM CNQX and 50 μM PTX. Representative example of eight cells, spikes are truncated at +20 mV. Firing before EtOH 1.07±0.22 Hz, after=7.18±1.58 Hz, paired t -test: t =3.757, d.f.=7, P =0.0071. ( c ) Effect of 50 mM of EtOH on membrane potential in whole-cell recordings, under synaptic blockade with 1 μM TTX. Top , membrane potential response; Bottom, responses to hyperpolarizing current injections (resting potentials artificially aligned to dotted line to facilitate comparison). Representative example of eight cells. Mean depolarization was 9.38±0.82 mV, one-sample t -test: t =11.4, d.f.=7, P
    Figure Legend Snippet: Effects of EtOH on membrane electrical properties of Agrp neurons. ( a ) Effect of 50 mM EtOH on spike rate in cell-attached recordings. Representative example of six cells. Firing before EtOH=1.85±0.35 Hz, after=3.72±0.52 Hz, paired t -test: t =4.613, d.f.=5, P =0.0058. ( b ) Effect of 50 mM EtOH on membrane potential, under synaptic blockade with of 10 μM CNQX and 50 μM PTX. Representative example of eight cells, spikes are truncated at +20 mV. Firing before EtOH 1.07±0.22 Hz, after=7.18±1.58 Hz, paired t -test: t =3.757, d.f.=7, P =0.0071. ( c ) Effect of 50 mM of EtOH on membrane potential in whole-cell recordings, under synaptic blockade with 1 μM TTX. Top , membrane potential response; Bottom, responses to hyperpolarizing current injections (resting potentials artificially aligned to dotted line to facilitate comparison). Representative example of eight cells. Mean depolarization was 9.38±0.82 mV, one-sample t -test: t =11.4, d.f.=7, P

    Techniques Used:

    23) Product Images from "Response of Cultured Neuronal Network Activity After High-Intensity Power Frequency Magnetic Field Exposure"

    Article Title: Response of Cultured Neuronal Network Activity After High-Intensity Power Frequency Magnetic Field Exposure

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2018.00189

    Effect of hPF-MF exposure on the autonomous activity of pacemaker-like neurons. (A) Screening of autonomous activity using MEAs and the pharmacological method. Even after D-AP5 and CNQX application, autonomous activity was detected in a few electrodes. (B) Detection of single pacemaker-like neuronal activity. Spontaneous activity with a frequency of 4–10 Hz, which is found in inhibitory neurons, was selected in some electrodes, then the effects of hPF-MF exposure on autonomous activity were evaluated. (C) Evaluation of the spike frequency after 400 mT hPF-MF exposure on the various pacemaker-like neurons. All autonomous activity detected from different cultures was reduced after 400 mT hPF-MF exposure. (D) Comparison of spike frequency before and after 400 mT hPF-MF exposure. N = 6 independent pacemaker-like neurons, ± SD, * P
    Figure Legend Snippet: Effect of hPF-MF exposure on the autonomous activity of pacemaker-like neurons. (A) Screening of autonomous activity using MEAs and the pharmacological method. Even after D-AP5 and CNQX application, autonomous activity was detected in a few electrodes. (B) Detection of single pacemaker-like neuronal activity. Spontaneous activity with a frequency of 4–10 Hz, which is found in inhibitory neurons, was selected in some electrodes, then the effects of hPF-MF exposure on autonomous activity were evaluated. (C) Evaluation of the spike frequency after 400 mT hPF-MF exposure on the various pacemaker-like neurons. All autonomous activity detected from different cultures was reduced after 400 mT hPF-MF exposure. (D) Comparison of spike frequency before and after 400 mT hPF-MF exposure. N = 6 independent pacemaker-like neurons, ± SD, * P

    Techniques Used: Activity Assay

    24) Product Images from "Gastrin-releasing peptide receptor mediates the excitation of preoptic GABAergic neurons by bombesin"

    Article Title: Gastrin-releasing peptide receptor mediates the excitation of preoptic GABAergic neurons by bombesin

    Journal: Neuroscience letters

    doi: 10.1016/j.neulet.2016.09.045

    GRP activates a cationic current in preoptic GABAergic neurons A. Responses to GRP (0.3 μM) recorded at three holding potentials. TTX (1 μM) and blockers of synaptic inputs (20 μM CNQX, 50 μM AP-5 and 5 μM gabazine) were present in the external solution. B. I-V relationship for the peak inward current activated by GRP (0.3 μM) recorded with K + pipette solution (filled circles) and Cs + pipette solution (open circles). Each point represents the average from n=5 neurons. C. Responses to GRP (0.3 μM) recorded in control aCSF (left trace) and in NMDG external solution (right trace) at -60 mV holding potential. TTX (1 μM) was present in all external solutions.
    Figure Legend Snippet: GRP activates a cationic current in preoptic GABAergic neurons A. Responses to GRP (0.3 μM) recorded at three holding potentials. TTX (1 μM) and blockers of synaptic inputs (20 μM CNQX, 50 μM AP-5 and 5 μM gabazine) were present in the external solution. B. I-V relationship for the peak inward current activated by GRP (0.3 μM) recorded with K + pipette solution (filled circles) and Cs + pipette solution (open circles). Each point represents the average from n=5 neurons. C. Responses to GRP (0.3 μM) recorded in control aCSF (left trace) and in NMDG external solution (right trace) at -60 mV holding potential. TTX (1 μM) was present in all external solutions.

    Techniques Used: Transferring

    25) Product Images from "Selectively driving cholinergic fibers optically in the thalamic reticular nucleus promotes sleep"

    Article Title: Selectively driving cholinergic fibers optically in the thalamic reticular nucleus promotes sleep

    Journal: eLife

    doi: 10.7554/eLife.10382

    Optically induced EPSCs were blocked by MLA. ( A ) TTX (1 μM) blocked the induced EPSCs in PV neurons evoked by optical stimulation of cholinergic fibers in the TRN (n = 6). ( B ) CNQX (20 μM), AP5 (50 μM), and PTX (100 μM) together failed to block the induced EPSCs (n = 6). ( C ) DHβE (5, 10 nM) only partially blocked the evoked EPSCs. ( D,E ) MLA (5 nM) completely blocked the evoked EPSCs and bursts of APs (n = 6). For voltage clamp recording in ( A, B, C, D ), the membrane potential was held at -70 mV. ( F ) Immunostaining with an antibody specific for α7-nAChRs showed substantial levels of the receptors on PV -positive neurons. Scale bar: 20 μm (n = 4). Data represent mean ± SEM (*p
    Figure Legend Snippet: Optically induced EPSCs were blocked by MLA. ( A ) TTX (1 μM) blocked the induced EPSCs in PV neurons evoked by optical stimulation of cholinergic fibers in the TRN (n = 6). ( B ) CNQX (20 μM), AP5 (50 μM), and PTX (100 μM) together failed to block the induced EPSCs (n = 6). ( C ) DHβE (5, 10 nM) only partially blocked the evoked EPSCs. ( D,E ) MLA (5 nM) completely blocked the evoked EPSCs and bursts of APs (n = 6). For voltage clamp recording in ( A, B, C, D ), the membrane potential was held at -70 mV. ( F ) Immunostaining with an antibody specific for α7-nAChRs showed substantial levels of the receptors on PV -positive neurons. Scale bar: 20 μm (n = 4). Data represent mean ± SEM (*p

    Techniques Used: Blocking Assay, Immunostaining

    26) Product Images from "Enhanced NMDA receptor NR1 phosphorylation and neuronal activity in the arcuate nucleus of hypothalamus following peripheral inflammation"

    Article Title: Enhanced NMDA receptor NR1 phosphorylation and neuronal activity in the arcuate nucleus of hypothalamus following peripheral inflammation

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2010.190

    MK-801 (NMDA receptor antagonist) and CNQX (non-NMDA receptor antagonist) inhibit spontaneous discharge of ARC neurons in inflamed rats. (A) A significant decrease in neuronal discharge frequency was observed following MK-801 application (300 μmol/L) in inflamed rats ( n =11; P
    Figure Legend Snippet: MK-801 (NMDA receptor antagonist) and CNQX (non-NMDA receptor antagonist) inhibit spontaneous discharge of ARC neurons in inflamed rats. (A) A significant decrease in neuronal discharge frequency was observed following MK-801 application (300 μmol/L) in inflamed rats ( n =11; P

    Techniques Used:

    27) Product Images from "Pharmacodynamics of the Glutamate Receptor Antagonists in the Rat Barrel Cortex"

    Article Title: Pharmacodynamics of the Glutamate Receptor Antagonists in the Rat Barrel Cortex

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2018.00698

    Sensory evoked activity resistant to ionotropic glutamate receptor antagonists in the thalamorecipient cortical layers. (A) Top , Stimulus-triggered LFP averages ( n = 30 responses; black traces) of sensory responses evoked by principal whisker deflection across cortical depths of the corresponding barrel column overlaid on color-coded current source density plot (CSD) in control, 40 min after 500 μM CNQX and 2 mM dAPV application, and 10 min after consecutive 2% lidocaine application. (B) Corresponding stimulus-triggered averages for MUA ( n = 30 responses). Stimulus onset on (A,B) is indicated by the black arrowhead and vertical magenta line. The cortical layer borders are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right. (C) The time course of early evoked L4 MUA suppression by 2% lidocaine epipially applied (gray bar above the plot) after the full blockade of SEP by CNQX/dAPV. The shaded area around the curve show SE bands ( n = 4 animals). The white circle on the curve indicates a half-maximal effect time (T½). (D) Averaged L4 LFP and MUA in control, in the presence of CNQX and dAPV, and after lidocaine application. Before averaging, individual LFP and MUA curves were aligned by the onset of SEP. Black arrowhead with an error bar above the dashed magenta line shows a mean time with SE between the stimulus and SEP onset. (E) Normalized mean values of SEP amplitude and early MUA peak in cortical L4 in control, in the presence of CNQX and dAPV, and after lidocaine application. (D,E) Show averaged data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.
    Figure Legend Snippet: Sensory evoked activity resistant to ionotropic glutamate receptor antagonists in the thalamorecipient cortical layers. (A) Top , Stimulus-triggered LFP averages ( n = 30 responses; black traces) of sensory responses evoked by principal whisker deflection across cortical depths of the corresponding barrel column overlaid on color-coded current source density plot (CSD) in control, 40 min after 500 μM CNQX and 2 mM dAPV application, and 10 min after consecutive 2% lidocaine application. (B) Corresponding stimulus-triggered averages for MUA ( n = 30 responses). Stimulus onset on (A,B) is indicated by the black arrowhead and vertical magenta line. The cortical layer borders are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right. (C) The time course of early evoked L4 MUA suppression by 2% lidocaine epipially applied (gray bar above the plot) after the full blockade of SEP by CNQX/dAPV. The shaded area around the curve show SE bands ( n = 4 animals). The white circle on the curve indicates a half-maximal effect time (T½). (D) Averaged L4 LFP and MUA in control, in the presence of CNQX and dAPV, and after lidocaine application. Before averaging, individual LFP and MUA curves were aligned by the onset of SEP. Black arrowhead with an error bar above the dashed magenta line shows a mean time with SE between the stimulus and SEP onset. (E) Normalized mean values of SEP amplitude and early MUA peak in cortical L4 in control, in the presence of CNQX and dAPV, and after lidocaine application. (D,E) Show averaged data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.

    Techniques Used: Activity Assay, Whisker Assay

    The time course of CNQX and dAPV effect on sensory evoked responses through the depth of the barrel cortex. The decrease in SEP slope (A) and MUA during SEP (B) in different layers of cortical barrel column following application of CNQX and dAPV. The application time is shown with a gray bar above the plot. White circles on the curves indicate a half-maximal effect time (T½). The colored areas surrounding each curve show SE bands. (C) Statistical data on T½ of CNQX and dAPV effect on SEP slope ( left ), early evoked MUA ( middle ) and late evoked MUA ( right ) across cortical layers. The open circles correspond to individual experiments. The medians of boxplots are shown by red lines. (A–C) , pooled data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.
    Figure Legend Snippet: The time course of CNQX and dAPV effect on sensory evoked responses through the depth of the barrel cortex. The decrease in SEP slope (A) and MUA during SEP (B) in different layers of cortical barrel column following application of CNQX and dAPV. The application time is shown with a gray bar above the plot. White circles on the curves indicate a half-maximal effect time (T½). The colored areas surrounding each curve show SE bands. (C) Statistical data on T½ of CNQX and dAPV effect on SEP slope ( left ), early evoked MUA ( middle ) and late evoked MUA ( right ) across cortical layers. The open circles correspond to individual experiments. The medians of boxplots are shown by red lines. (A–C) , pooled data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.

    Techniques Used:

    The CNQX and dAPV action on spontaneous activity through the depth of the barrel column. (A) Example traces of 16-channel recording of barrel cortex activity in control ( top ), 15 min after epipial application of 500 μM CNQX and 2 mM dAPV ( middle ), and 60 min after drug application ( bottom ). Red bars indicate detected spikes (MUA). (B) Corresponding current source density (CSD, upper panels ) and MUA density ( lower panels ) plots of averaged up-states recorded in control and after drug application (150 events were averaged for each condition). LFP traces (black) are overlaid on color-coded CSD plots. The cortical layer borders on (A,B) are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right.
    Figure Legend Snippet: The CNQX and dAPV action on spontaneous activity through the depth of the barrel column. (A) Example traces of 16-channel recording of barrel cortex activity in control ( top ), 15 min after epipial application of 500 μM CNQX and 2 mM dAPV ( middle ), and 60 min after drug application ( bottom ). Red bars indicate detected spikes (MUA). (B) Corresponding current source density (CSD, upper panels ) and MUA density ( lower panels ) plots of averaged up-states recorded in control and after drug application (150 events were averaged for each condition). LFP traces (black) are overlaid on color-coded CSD plots. The cortical layer borders on (A,B) are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right.

    Techniques Used: Activity Assay

    The CNQX and dAPV effect on sensory evoked fast oscillations (FO) in cortical layer 4. (A) Three representative traces (after high-pass filtering, 200–1000 Hz) show fast L4 LFP and MUA oscillations evoked by sensory stimulation. Red bars overlaid on the LFP traces are detected spikes (MUA). Stimulus onset is indicated by the arrowhead. Corresponding PSTH of evoked MUA averaged over 30 responses is shown under the LFP traces. (B) Top , averaged power spectrum of evoked L4 MUA before and during CNQX/dAPV application (application time is shown with a white bar). Bottom , the time-course of sensory-evoked (early + late) MUA suppression by CNQX/dAPV. Shaded area shows SE bands. Data were averaged over 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.
    Figure Legend Snippet: The CNQX and dAPV effect on sensory evoked fast oscillations (FO) in cortical layer 4. (A) Three representative traces (after high-pass filtering, 200–1000 Hz) show fast L4 LFP and MUA oscillations evoked by sensory stimulation. Red bars overlaid on the LFP traces are detected spikes (MUA). Stimulus onset is indicated by the arrowhead. Corresponding PSTH of evoked MUA averaged over 30 responses is shown under the LFP traces. (B) Top , averaged power spectrum of evoked L4 MUA before and during CNQX/dAPV application (application time is shown with a white bar). Bottom , the time-course of sensory-evoked (early + late) MUA suppression by CNQX/dAPV. Shaded area shows SE bands. Data were averaged over 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV.

    Techniques Used:

    The statistical data on CNQX and dAPV action on spontaneous barrel cortex activity. The decrease in up-states amplitude (A) , MUA density during up-states (B) , and up-states frequency (C) in different layers of the cortical barrel column following application of CNQX/dAPV (application time shown with gray bar above each plot). The colored areas surrounding each curve show SE bands. (D) The up-state duration distribution in control and after the drugs application. (A–D) Show averaged data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV. (E) Group data on normalized MUA density in different cortical layers during a steady state of CNQX and dAPV effect. (F) Group data on up-states frequency in control and after CNQX/dAPV application. The open circles on (E,F) correspond to individual experiments, and the red line on boxplots is a median. ∗ p
    Figure Legend Snippet: The statistical data on CNQX and dAPV action on spontaneous barrel cortex activity. The decrease in up-states amplitude (A) , MUA density during up-states (B) , and up-states frequency (C) in different layers of the cortical barrel column following application of CNQX/dAPV (application time shown with gray bar above each plot). The colored areas surrounding each curve show SE bands. (D) The up-state duration distribution in control and after the drugs application. (A–D) Show averaged data from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV. (E) Group data on normalized MUA density in different cortical layers during a steady state of CNQX and dAPV effect. (F) Group data on up-states frequency in control and after CNQX/dAPV application. The open circles on (E,F) correspond to individual experiments, and the red line on boxplots is a median. ∗ p

    Techniques Used: Activity Assay

    Depth profile of CNQX and dAPV action on barrel cortex activity. (A) Top , Stimulus-triggered LFP averages (black traces) of sensory responses evoked by principal whisker deflection across cortical depths of corresponding barrel column overlaid on color-coded current source density plot (CSD) in control, 7 and 25 min after 500 μM CNQX and 2 mM dAPV application (25 responses were averaged for each CSD plot). Bottom , corresponding stimulus-triggered averages for MUA. Stimulus onset is indicated by the black arrowhead and vertical magenta line. The cortical layer borders are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right. Red square brackets indicate the electrodes of the recording that were used for building group data plots on panel (B) . The gray bars above MUA density plots limit the time windows for early and late MUA density calculation. (B) Group data (from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV) on the magnitude of CNQX and dAPV effect on early and late components of the sensory-evoked MUA, spontaneous MUA and SEP slope at the beginning of drug application (4–9 min, the time point corresponding to SEP slope reduction by 20% of control values) and at the end of 1 h of application (45–60 min). Note the gradual development of CNQX/dAPV effect from superficial toward deep cortical layers, and the persistence of early evoked L4 spikes after full blockade of SEP and nearly complete block of evoked MUA in L2/3 and L5a.
    Figure Legend Snippet: Depth profile of CNQX and dAPV action on barrel cortex activity. (A) Top , Stimulus-triggered LFP averages (black traces) of sensory responses evoked by principal whisker deflection across cortical depths of corresponding barrel column overlaid on color-coded current source density plot (CSD) in control, 7 and 25 min after 500 μM CNQX and 2 mM dAPV application (25 responses were averaged for each CSD plot). Bottom , corresponding stimulus-triggered averages for MUA. Stimulus onset is indicated by the black arrowhead and vertical magenta line. The cortical layer borders are shown left of the CSD and MUA density plots, and recording electrode depths are shown on the right. Red square brackets indicate the electrodes of the recording that were used for building group data plots on panel (B) . The gray bars above MUA density plots limit the time windows for early and late MUA density calculation. (B) Group data (from 11 animals and two concentrations, 170 μM CNQX/700 μM dAPV and 0.5 mM CNQX/2 mM dAPV) on the magnitude of CNQX and dAPV effect on early and late components of the sensory-evoked MUA, spontaneous MUA and SEP slope at the beginning of drug application (4–9 min, the time point corresponding to SEP slope reduction by 20% of control values) and at the end of 1 h of application (45–60 min). Note the gradual development of CNQX/dAPV effect from superficial toward deep cortical layers, and the persistence of early evoked L4 spikes after full blockade of SEP and nearly complete block of evoked MUA in L2/3 and L5a.

    Techniques Used: Activity Assay, Whisker Assay, Blocking Assay

    Pharmacokinetics of CNQX and dAPV after epipial application. (A) Development of the inhibitory effect of CNQX and dAPV on SEP amplitude across cortical layers over 1 h of application (averaged data from 11 animals). (B) Estimation of CNQX concentration dynamics in different cortical layers after epipial drug application. Instant values of CNQX concentration were calculated based on the concentration dependence of CNQX effect on the fast EP SCs with a K d of 0.4 μM and Hill coefficient of 1 ( Honore et al., 1988 ; Llano et al., 1991 ). (C) The time points where epipially applied CNQX (170–500 μM) attains K d values of 0.4 and 10 μM at different depths of the cortical column.
    Figure Legend Snippet: Pharmacokinetics of CNQX and dAPV after epipial application. (A) Development of the inhibitory effect of CNQX and dAPV on SEP amplitude across cortical layers over 1 h of application (averaged data from 11 animals). (B) Estimation of CNQX concentration dynamics in different cortical layers after epipial drug application. Instant values of CNQX concentration were calculated based on the concentration dependence of CNQX effect on the fast EP SCs with a K d of 0.4 μM and Hill coefficient of 1 ( Honore et al., 1988 ; Llano et al., 1991 ). (C) The time points where epipially applied CNQX (170–500 μM) attains K d values of 0.4 and 10 μM at different depths of the cortical column.

    Techniques Used: Concentration Assay

    28) Product Images from "Recurrent Dendrodendritic Inhibition of Accessory Olfactory Bulb Mitral Cells Requires Activation of Group I Metabotropic Glutamate Receptors"

    Article Title: Recurrent Dendrodendritic Inhibition of Accessory Olfactory Bulb Mitral Cells Requires Activation of Group I Metabotropic Glutamate Receptors

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0613-07.2007

    The increase in IPSC rate with DHPG addition occurs via a mechanism presynaptic to mitral cells. A , Sample sweep in which APV (50 μ m ) and CNQX (20 μ m ) were bath applied for 10 min before addition of 20 μ m DHPG (baseline period
    Figure Legend Snippet: The increase in IPSC rate with DHPG addition occurs via a mechanism presynaptic to mitral cells. A , Sample sweep in which APV (50 μ m ) and CNQX (20 μ m ) were bath applied for 10 min before addition of 20 μ m DHPG (baseline period

    Techniques Used:

    29) Product Images from "FE65 Proteins Regulate NMDA Receptor Activation-induced Amyloid Precursor Protein Processing"

    Article Title: FE65 Proteins Regulate NMDA Receptor Activation-induced Amyloid Precursor Protein Processing

    Journal: Journal of neurochemistry

    doi: 10.1111/j.1471-4159.2011.07419.x

    Cytotoxic NMDA levels abrogate the differential effect of FE65 protein deficiencies on NMDAR-activation dependent APP processing (a) WT and FE65/FE65L1 DKO cortical neurons (DIV 13) were exposed to NMDA (100 μM) for the indicated times. MK801, an NMDA antagonist, was co-incubated with NMDA for the 2.5 hr treatment group. (b) WT or DKO neurons (DIV13) were exposed to the indicated doses of NMDA for 3 hr. (c) WT neurons (DIV 13) were exposed to NMDA (100 μM) or AMPA (10 μM) alone, or in combination with MK801 (10 μM) or CNQX (10 μM) for 3 hr. APP695, APP-CTFs and HMW species were detected by Western blot analysis using the APP C-terminal antibody (A8717). p-APP, pThr668-APP was detected on the same blot with a pThr668APP specific antibody and β-actin was used as a loading control.
    Figure Legend Snippet: Cytotoxic NMDA levels abrogate the differential effect of FE65 protein deficiencies on NMDAR-activation dependent APP processing (a) WT and FE65/FE65L1 DKO cortical neurons (DIV 13) were exposed to NMDA (100 μM) for the indicated times. MK801, an NMDA antagonist, was co-incubated with NMDA for the 2.5 hr treatment group. (b) WT or DKO neurons (DIV13) were exposed to the indicated doses of NMDA for 3 hr. (c) WT neurons (DIV 13) were exposed to NMDA (100 μM) or AMPA (10 μM) alone, or in combination with MK801 (10 μM) or CNQX (10 μM) for 3 hr. APP695, APP-CTFs and HMW species were detected by Western blot analysis using the APP C-terminal antibody (A8717). p-APP, pThr668-APP was detected on the same blot with a pThr668APP specific antibody and β-actin was used as a loading control.

    Techniques Used: Activation Assay, Incubation, Western Blot

    30) Product Images from "NMDA Receptor Activation and Calpain Contribute to Disruption of Dendritic Spines by the Stress Neuropeptide CRH"

    Article Title: NMDA Receptor Activation and Calpain Contribute to Disruption of Dendritic Spines by the Stress Neuropeptide CRH

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1445-13.2013

    CRH-induced spine loss requires NMDA receptor activation, but not AMPA receptor activation. To distinguish the roles of NMDA- and AMPA-type receptors, neurons were exposed to CRH in the presence of selective blockers, APV and CNQX, respectively. A , Example of dendrites exposed to CRH in the presence of APV + CNQX. The combined antagonists prevented CRH-induced spine loss. B , Graph quantifying CRH spine loss in the presence of both glutamate receptor antagonists ( F (2,180) = 64.22, p
    Figure Legend Snippet: CRH-induced spine loss requires NMDA receptor activation, but not AMPA receptor activation. To distinguish the roles of NMDA- and AMPA-type receptors, neurons were exposed to CRH in the presence of selective blockers, APV and CNQX, respectively. A , Example of dendrites exposed to CRH in the presence of APV + CNQX. The combined antagonists prevented CRH-induced spine loss. B , Graph quantifying CRH spine loss in the presence of both glutamate receptor antagonists ( F (2,180) = 64.22, p

    Techniques Used: Activation Assay

    31) Product Images from "Comparison of Metal and Metal Oxide Media for Phosphopeptide Enrichment Prior to Mass Spectrometric Analyses"

    Article Title: Comparison of Metal and Metal Oxide Media for Phosphopeptide Enrichment Prior to Mass Spectrometric Analyses

    Journal: Journal of the American Society for Mass Spectrometry

    doi: 10.1016/j.jasms.2010.06.005

    MALDI mass spectrum of an α- and β-casein tryptic digest resulting from enrichment using magnetic titanium dioxide beads (Phos-Trap kit). The ion labeled with an asterisk (*) corresponds in mass to a previously reported monophosphorylated
    Figure Legend Snippet: MALDI mass spectrum of an α- and β-casein tryptic digest resulting from enrichment using magnetic titanium dioxide beads (Phos-Trap kit). The ion labeled with an asterisk (*) corresponds in mass to a previously reported monophosphorylated

    Techniques Used: Labeling

    MALDI mass spectra of an α- and β-casein tryptic digest resulting from A) titanium dioxide NuTip enrichment, B) zirconium dioxide NuTip enrichment, and C) a mixed resin tip enrichment consisting of zirconium and titanium dioxide. The ion
    Figure Legend Snippet: MALDI mass spectra of an α- and β-casein tryptic digest resulting from A) titanium dioxide NuTip enrichment, B) zirconium dioxide NuTip enrichment, and C) a mixed resin tip enrichment consisting of zirconium and titanium dioxide. The ion

    Techniques Used:

    MALDI/MS of an α- and β-casein tryptic digest following A) Fe-NTA IMAC CRC enrichment, and B) titanium dioxide (GL Sciences) MOAC CRC enrichment. The ion labeled with an asterisk (*) corresponds in mass to a previously reported monophosphorylated
    Figure Legend Snippet: MALDI/MS of an α- and β-casein tryptic digest following A) Fe-NTA IMAC CRC enrichment, and B) titanium dioxide (GL Sciences) MOAC CRC enrichment. The ion labeled with an asterisk (*) corresponds in mass to a previously reported monophosphorylated

    Techniques Used: Mass Spectrometry, Labeling

    32) Product Images from "The adipocyte differentiation protein APMAP is an endogenous suppressor of Aβ production in the brain"

    Article Title: The adipocyte differentiation protein APMAP is an endogenous suppressor of Aβ production in the brain

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddu449

    APMAP controls the levels and the stability of APP-CTFs. ( A ) siRNA knockdown of APMAP in HeLa cells quantitatively and qualitatively impacts APP-CTFs. Reduced APMAP expression correlates with increased APP-CTFs and the formation of three forms of APP-CTFα (CTFα1, α2 and α3), as revealed on a Tris-Tricine urea gel (left panel). This phenotype is further amplified in the presence of 1 µ m GSI DAPT (right panel). Biological triplicates are shown. ( B ) MALDI-TOF mass spectrometric analysis of APP-CTFα1, α2 and α3 immunoprecipitated from DAPT-treated cells with siRNA-reduced APMAP expression. ( C ) Peptide sequences of N-terminal truncated APP-CTFα1 (C80), -CTFα2 (C74) and -CTFα3 (C71).
    Figure Legend Snippet: APMAP controls the levels and the stability of APP-CTFs. ( A ) siRNA knockdown of APMAP in HeLa cells quantitatively and qualitatively impacts APP-CTFs. Reduced APMAP expression correlates with increased APP-CTFs and the formation of three forms of APP-CTFα (CTFα1, α2 and α3), as revealed on a Tris-Tricine urea gel (left panel). This phenotype is further amplified in the presence of 1 µ m GSI DAPT (right panel). Biological triplicates are shown. ( B ) MALDI-TOF mass spectrometric analysis of APP-CTFα1, α2 and α3 immunoprecipitated from DAPT-treated cells with siRNA-reduced APMAP expression. ( C ) Peptide sequences of N-terminal truncated APP-CTFα1 (C80), -CTFα2 (C74) and -CTFα3 (C71).

    Techniques Used: Expressing, Amplification, Immunoprecipitation

    33) Product Images from "Density Functional Theory Analysis of Deltamethrin and Its Determination in Strawberry by Surface Enhanced Raman Spectroscopy"

    Article Title: Density Functional Theory Analysis of Deltamethrin and Its Determination in Strawberry by Surface Enhanced Raman Spectroscopy

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    doi: 10.3390/molecules23061458

    ( a ) 500–1800 cm −1 SERS spectra of different concentrations of deltamethrin in strawberry; ( b ) the SERS spectra of five concentrations of deltamethrin in strawberry.
    Figure Legend Snippet: ( a ) 500–1800 cm −1 SERS spectra of different concentrations of deltamethrin in strawberry; ( b ) the SERS spectra of five concentrations of deltamethrin in strawberry.

    Techniques Used:

    The RS of deltamethrin: ( a ) the theory calculation by density functional theory; ( b ) RS of deltamethrin solid; ( c ) SERS of deltamethrin solution.
    Figure Legend Snippet: The RS of deltamethrin: ( a ) the theory calculation by density functional theory; ( b ) RS of deltamethrin solid; ( c ) SERS of deltamethrin solution.

    Techniques Used: Functional Assay

    The SERS of deltamethrin solution based on two nanoparticles: (a) AgNPs; (b) AuNPs; the SERS of two nanoparticles; (c) AgNPs; (d) AuNPs; the RS of acetonitrile; (e) acetonitrile.
    Figure Legend Snippet: The SERS of deltamethrin solution based on two nanoparticles: (a) AgNPs; (b) AuNPs; the SERS of two nanoparticles; (c) AgNPs; (d) AuNPs; the RS of acetonitrile; (e) acetonitrile.

    Techniques Used:

    Raman shift deviation between the Raman characteristic peaks of deltamethrin and the Raman characteristic peaks calculated by DFT.
    Figure Legend Snippet: Raman shift deviation between the Raman characteristic peaks of deltamethrin and the Raman characteristic peaks calculated by DFT.

    Techniques Used:

    Simulated molecular structure of deltamethrin by DFT.
    Figure Legend Snippet: Simulated molecular structure of deltamethrin by DFT.

    Techniques Used:

    34) Product Images from "Role of Molybdenum-Containing Enzymes in the Biotransformation of the Novel Ghrelin Receptor Inverse Agonist PF-5190457: A Reverse Translational Bed-to-Bench Approach"

    Article Title: Role of Molybdenum-Containing Enzymes in the Biotransformation of the Novel Ghrelin Receptor Inverse Agonist PF-5190457: A Reverse Translational Bed-to-Bench Approach

    Journal: Drug Metabolism and Disposition

    doi: 10.1124/dmd.119.087015

    Hydroxy metabolite (PF-6870961) formation in the presence of inhibitors (percentage of control) at various concentrations of raloxifene (A) and febuxostat (B) with 25 µ M PF-5190457 in HLC incubations.
    Figure Legend Snippet: Hydroxy metabolite (PF-6870961) formation in the presence of inhibitors (percentage of control) at various concentrations of raloxifene (A) and febuxostat (B) with 25 µ M PF-5190457 in HLC incubations.

    Techniques Used:

    35) Product Images from "Chemoresistance to Concanamycin A1 in Human Oral Squamous Cell Carcinoma Is Attenuated by an HDAC Inhibitor Partly via Suppression of Bcl-2 Expression"

    Article Title: Chemoresistance to Concanamycin A1 in Human Oral Squamous Cell Carcinoma Is Attenuated by an HDAC Inhibitor Partly via Suppression of Bcl-2 Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0080998

    Expression of V-ATPase subunit genes and pro- and anti-apoptotic genes in OSCC cells. Expression of V-ATPase subunit genes and pro- and anti-apoptotic genes was compared in OSCC cell lines, MISK81-5, SAS, HSC-4 and SQUU-B cells, without CMA treatment. The ATP6V0A3 ( A ) ATP6V0A4 ( B ) and ATP6V1C1 ( C ) mRNA expression levels were analyzed by qRT-PCR. The Bax ( D ) and Bcl-2 ( E ) mRNA expression levels were also analyzed. The Bax/Bcl-2 ratio ( F ) in each OSCC cell line was calculated based on the qRT-PCR data. GAPDH was used as an internal control for the experiment. Columns , means of at least triplicate experiments; bars, SD. * and ** denote p
    Figure Legend Snippet: Expression of V-ATPase subunit genes and pro- and anti-apoptotic genes in OSCC cells. Expression of V-ATPase subunit genes and pro- and anti-apoptotic genes was compared in OSCC cell lines, MISK81-5, SAS, HSC-4 and SQUU-B cells, without CMA treatment. The ATP6V0A3 ( A ) ATP6V0A4 ( B ) and ATP6V1C1 ( C ) mRNA expression levels were analyzed by qRT-PCR. The Bax ( D ) and Bcl-2 ( E ) mRNA expression levels were also analyzed. The Bax/Bcl-2 ratio ( F ) in each OSCC cell line was calculated based on the qRT-PCR data. GAPDH was used as an internal control for the experiment. Columns , means of at least triplicate experiments; bars, SD. * and ** denote p

    Techniques Used: Expressing, Quantitative RT-PCR

    Effects of CMA on the expression of pro- and anti-apoptotic genes and proteins in OSCC cells. OSCC cell lines, MISK81-5, SAS, HSC-4 and SQUU-B cells, were treated with or without CMA. The Bax ( A ) and Bcl-2 ( B ) mRNA expression levels were analyzed at 24 hr after CMA treatment by qRT-PCR. The Bax/Bcl-2 ratio ( C ) in each OSCC cell line was calculated based on the qRT-PCR data. GAPDH was used as internal control for the experiment. The Bax and Bcl-2 protein expression levels in the SAS ( D ) and SQUU-B ( E ) cell lines were analyzed at 24 hr by a Western blot analysis. The Bax level, Bcl-2 level and Bax/Bcl-2 ratio was calculated based on the intensity of the bands in the SAS ( F ) and SQUU-B ( G ) cell lines. β-actin was used as an internal control for the experiment. Columns , means of at least triplicate experiments; bars, SD. * and ** denote p
    Figure Legend Snippet: Effects of CMA on the expression of pro- and anti-apoptotic genes and proteins in OSCC cells. OSCC cell lines, MISK81-5, SAS, HSC-4 and SQUU-B cells, were treated with or without CMA. The Bax ( A ) and Bcl-2 ( B ) mRNA expression levels were analyzed at 24 hr after CMA treatment by qRT-PCR. The Bax/Bcl-2 ratio ( C ) in each OSCC cell line was calculated based on the qRT-PCR data. GAPDH was used as internal control for the experiment. The Bax and Bcl-2 protein expression levels in the SAS ( D ) and SQUU-B ( E ) cell lines were analyzed at 24 hr by a Western blot analysis. The Bax level, Bcl-2 level and Bax/Bcl-2 ratio was calculated based on the intensity of the bands in the SAS ( F ) and SQUU-B ( G ) cell lines. β-actin was used as an internal control for the experiment. Columns , means of at least triplicate experiments; bars, SD. * and ** denote p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    36) Product Images from "Amphiphilic Fluorinated Block Copolymer Synthesized by RAFT Polymerization for Graphene Dispersions"

    Article Title: Amphiphilic Fluorinated Block Copolymer Synthesized by RAFT Polymerization for Graphene Dispersions

    Journal: Polymers

    doi: 10.3390/polym8030101

    Schematic drawing of the mechanism for syntheses of ( a ) PTFEMA macro-RAFT agent (PTFEMA n -CTP) and ( b ) PTFEMA n - b -PVP m block copolymer. CTP: 4-Cyano-4-(phenylcarbonothioylthio)pentanoic acid; TFEMA: 2,2,2-Trifluoroethyl methacrylate; AIBN: 2,2′-Azobisisobutyronitrile; VP: 4-Vinyl pyridine; PTFEMA-b-PVP: Poly(2,2,2-trifluoroethyl methacrylate)- block -poly(4-vinyl pyridine). RAFT: Reversible addition-fragmentation chain transfer.
    Figure Legend Snippet: Schematic drawing of the mechanism for syntheses of ( a ) PTFEMA macro-RAFT agent (PTFEMA n -CTP) and ( b ) PTFEMA n - b -PVP m block copolymer. CTP: 4-Cyano-4-(phenylcarbonothioylthio)pentanoic acid; TFEMA: 2,2,2-Trifluoroethyl methacrylate; AIBN: 2,2′-Azobisisobutyronitrile; VP: 4-Vinyl pyridine; PTFEMA-b-PVP: Poly(2,2,2-trifluoroethyl methacrylate)- block -poly(4-vinyl pyridine). RAFT: Reversible addition-fragmentation chain transfer.

    Techniques Used: Blocking Assay

    37) Product Images from "Anti-Inflammatory Effects of Acupuncture Stimulation via the Vagus Nerve"

    Article Title: Anti-Inflammatory Effects of Acupuncture Stimulation via the Vagus Nerve

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0151882

    Regulation of c-Fos induction in the DVC by CNQX and PPADS. Following MAC and EAC in combination with focal administration of CNQX and/or PPADS into the DVC area, transverse sections through the caudal portion of the brainstem were subjected to immunofluoresence staining analyses for c-Fos production. Upper images are the representatives showing c-Fos signals in the brain sections and lower graphs show the quantitation of the protein signals in the field. Mean ± SEM (n = 4 independent experiment). *p
    Figure Legend Snippet: Regulation of c-Fos induction in the DVC by CNQX and PPADS. Following MAC and EAC in combination with focal administration of CNQX and/or PPADS into the DVC area, transverse sections through the caudal portion of the brainstem were subjected to immunofluoresence staining analyses for c-Fos production. Upper images are the representatives showing c-Fos signals in the brain sections and lower graphs show the quantitation of the protein signals in the field. Mean ± SEM (n = 4 independent experiment). *p

    Techniques Used: Staining, Quantitation Assay

    38) Product Images from "New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS"

    Article Title: New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21155265

    Networks built based on MS spectra obtained using ( a ) 2,5-dihydroxybenzoic acid (DHB) and ( b ) α-cyano-4-hydroxycinnamic acid (HCCA) matrices, in positive ionization mode; ( c ) DHB and ( d ) HCCA matrices, in negative ionization mode. Colored nodes: compounds, white rectangles: MS ions, e—compounds subjected to the electrochemical process, FRU: D-fructose, GAL: D-galactose, GLU: D-glucose, PI: D-pinitol, CI: L- chiro -inositol, and MI: myo -inositol.
    Figure Legend Snippet: Networks built based on MS spectra obtained using ( a ) 2,5-dihydroxybenzoic acid (DHB) and ( b ) α-cyano-4-hydroxycinnamic acid (HCCA) matrices, in positive ionization mode; ( c ) DHB and ( d ) HCCA matrices, in negative ionization mode. Colored nodes: compounds, white rectangles: MS ions, e—compounds subjected to the electrochemical process, FRU: D-fructose, GAL: D-galactose, GLU: D-glucose, PI: D-pinitol, CI: L- chiro -inositol, and MI: myo -inositol.

    Techniques Used:

    39) Product Images from "Synthesis and Characterization of Bioactive Acylpyrazolone Sulfanilamides and Their Transition Metal Complexes: Single Crystal Structure of 4-Benzoyl-3-methyl-1-phenyl-2-pyrazolin-5-one Sulfanilamide"

    Article Title: Synthesis and Characterization of Bioactive Acylpyrazolone Sulfanilamides and Their Transition Metal Complexes: Single Crystal Structure of 4-Benzoyl-3-methyl-1-phenyl-2-pyrazolin-5-one Sulfanilamide

    Journal: Bioinorganic Chemistry and Applications

    doi: 10.1155/2015/717089

    Synthesis of 4-acyl-3-methyl-1-phenyl-2-pyrazolin-5-one-sulfanilamide.
    Figure Legend Snippet: Synthesis of 4-acyl-3-methyl-1-phenyl-2-pyrazolin-5-one-sulfanilamide.

    Techniques Used:

    40) Product Images from "Breast cancer cells rely on environmental pyruvate to shape the metastatic niche"

    Article Title: Breast cancer cells rely on environmental pyruvate to shape the metastatic niche

    Journal: Nature

    doi: 10.1038/s41586-019-0977-x

    Pyruvate drives collagen hydroxylation ( a ) Pyruvate, lactate and glucose uptake/secretion in human MCF10A H-RAS V12 breast cancer spheroids treated with MCT2 inhibitor (α-cyano-hydroxycinnamic acid; 1.5mM). This data show that the used inhibitor impairs pyruvate uptake, but not lactate or glucose secretion and uptake, respectively. ( b-c ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (MCF10A H-RAS V12 , MCF7, HCC70) and mouse (4T1) breast cancer spheroids treated with a MCT2 inhibitor (α-cyano-hydroxycinnamic acid; 1.5 mM), or treated with the MPC inhibitor UK5099 (50 μM) in the presence of pyruvate. The number of biological replicates for each experiment was n=3. Error bars represent SD of mean from biological independent samples. Two-tailed unpaired student’s T-test.
    Figure Legend Snippet: Pyruvate drives collagen hydroxylation ( a ) Pyruvate, lactate and glucose uptake/secretion in human MCF10A H-RAS V12 breast cancer spheroids treated with MCT2 inhibitor (α-cyano-hydroxycinnamic acid; 1.5mM). This data show that the used inhibitor impairs pyruvate uptake, but not lactate or glucose secretion and uptake, respectively. ( b-c ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (MCF10A H-RAS V12 , MCF7, HCC70) and mouse (4T1) breast cancer spheroids treated with a MCT2 inhibitor (α-cyano-hydroxycinnamic acid; 1.5 mM), or treated with the MPC inhibitor UK5099 (50 μM) in the presence of pyruvate. The number of biological replicates for each experiment was n=3. Error bars represent SD of mean from biological independent samples. Two-tailed unpaired student’s T-test.

    Techniques Used: Two Tailed Test

    α-ketoglutarate metabolically regulates P4HA activity in cancer cells ( a ) Schematic representation of the metabolic regulation and carbon donor mechanisms by which α-ketoglutarate can regulate collagen hydroxylation. Solid lines indicate metabolite conversion, while dashed lines indicate metabolic regulation. Enzymes are depicted in italics. P4HA refers to collagen prolyl-4-hydroxlase. P5CS refers to pyrroline-5-carboxylate synthase. ( b-c ) Relative change in intracellular abundance of proline and hydroxylated collagen in MCF10A H-Ras V12 spheroids transduced with a lentiviral vector with shRNA for either P5CS (KD) or scrambled control sequence with or without cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM) in the presence or absence of pyruvate normalized to control condition. If the carbon donor mechanism occurs, it is expected that proline abundance decreases in P5CS knockdown spheroids and that they no longer respond to the α-ketoglutarate rescue upon pyruvate depletion. However, we observed that proline abundance did not significantly change in P5CS knockdown spheroids. Moreover, α-ketoglutarate addition still significantly increased hydroxylated collagen to a similar extent as pyruvate. n=3 (b); n=6 (control shRNA); n=3 (P5CS shRNA 1 and 2 (c)). ( d ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (myo)fibroblasts in presence or absence of pyruvate with or without cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM) and/or cell permeable succinate (dimethyl succinate; 1.5 mM). n=3 (human primary skin-derived fibroblasts); n=4 (human immortalized mammary and cancer associated myofibroblasts). ( e ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (myo)fibroblasts treated with the MCT2 inhibitor α-cyano-4-hydroxycinnamic acid (1.5 mM), the MPC inhibitor UK5099 (50 μM) or the transaminase inhibitor AOA (0.8 mM) in the presence of pyruvate. n=3. ( f ) Intracellular abundance of α-ketoglutarate (α-KG) in the presence or absence of pyruvate in human fibroblasts. n=3. Error bars represent SD of mean from biological independent samples. Two-tailed unpaired student’s T-test.
    Figure Legend Snippet: α-ketoglutarate metabolically regulates P4HA activity in cancer cells ( a ) Schematic representation of the metabolic regulation and carbon donor mechanisms by which α-ketoglutarate can regulate collagen hydroxylation. Solid lines indicate metabolite conversion, while dashed lines indicate metabolic regulation. Enzymes are depicted in italics. P4HA refers to collagen prolyl-4-hydroxlase. P5CS refers to pyrroline-5-carboxylate synthase. ( b-c ) Relative change in intracellular abundance of proline and hydroxylated collagen in MCF10A H-Ras V12 spheroids transduced with a lentiviral vector with shRNA for either P5CS (KD) or scrambled control sequence with or without cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM) in the presence or absence of pyruvate normalized to control condition. If the carbon donor mechanism occurs, it is expected that proline abundance decreases in P5CS knockdown spheroids and that they no longer respond to the α-ketoglutarate rescue upon pyruvate depletion. However, we observed that proline abundance did not significantly change in P5CS knockdown spheroids. Moreover, α-ketoglutarate addition still significantly increased hydroxylated collagen to a similar extent as pyruvate. n=3 (b); n=6 (control shRNA); n=3 (P5CS shRNA 1 and 2 (c)). ( d ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (myo)fibroblasts in presence or absence of pyruvate with or without cell permeable α-ketoglutarate (dimethyl 2-oxoglutarate; α-KG; 1.5 mM) and/or cell permeable succinate (dimethyl succinate; 1.5 mM). n=3 (human primary skin-derived fibroblasts); n=4 (human immortalized mammary and cancer associated myofibroblasts). ( e ) Hydroxylated collagen assessed via measurement of hydroxyproline (OH-proline) in human (myo)fibroblasts treated with the MCT2 inhibitor α-cyano-4-hydroxycinnamic acid (1.5 mM), the MPC inhibitor UK5099 (50 μM) or the transaminase inhibitor AOA (0.8 mM) in the presence of pyruvate. n=3. ( f ) Intracellular abundance of α-ketoglutarate (α-KG) in the presence or absence of pyruvate in human fibroblasts. n=3. Error bars represent SD of mean from biological independent samples. Two-tailed unpaired student’s T-test.

    Techniques Used: Metabolic Labelling, Activity Assay, Transduction, Plasmid Preparation, shRNA, Sequencing, Derivative Assay, Two Tailed Test

    Related Articles

    In Vivo:

    Article Title: Neural Stem Cells in the Adult Subventricular Zone Oxidize Fatty Acids to Produce Energy and Support Neurogenic Activity
    Article Snippet: .. Pharmacological Inhibition of Fatty Acid and Lactate Transport In Vivo Female C57B/6 mice between 5 and 7 weeks of age were injected intraperitoneally with 40 mg/kg etomoxir (Sigma, St Louis, MO) in PBS, 40 mg/kg alpha‐cyano‐4‐hydroxycinnamate (4‐CIN (Sigma, St Louis, MO)) in 1% methanol‐PBS, or with similar volumes of PBS. ..

    other:

    Article Title: Lactate, a putative survival factor for myeloma cells, is incorporated by myeloma cells through monocarboxylate transporters 1
    Article Snippet: Inhibitors Dichloroacetate (DCA) and α-cyano-4-hydroxycinnamic acid (CHC) were purchased from Sigma-Aldrich (St Louis, MO, USA), and dissolved in phosphate-buffered saline and dimethyl sulfoxide (DMSO), respectively.

    Inhibition:

    Article Title: Neural Stem Cells in the Adult Subventricular Zone Oxidize Fatty Acids to Produce Energy and Support Neurogenic Activity
    Article Snippet: .. Pharmacological Inhibition of Fatty Acid and Lactate Transport In Vivo Female C57B/6 mice between 5 and 7 weeks of age were injected intraperitoneally with 40 mg/kg etomoxir (Sigma, St Louis, MO) in PBS, 40 mg/kg alpha‐cyano‐4‐hydroxycinnamate (4‐CIN (Sigma, St Louis, MO)) in 1% methanol‐PBS, or with similar volumes of PBS. ..

    Injection:

    Article Title: Neural Stem Cells in the Adult Subventricular Zone Oxidize Fatty Acids to Produce Energy and Support Neurogenic Activity
    Article Snippet: .. Pharmacological Inhibition of Fatty Acid and Lactate Transport In Vivo Female C57B/6 mice between 5 and 7 weeks of age were injected intraperitoneally with 40 mg/kg etomoxir (Sigma, St Louis, MO) in PBS, 40 mg/kg alpha‐cyano‐4‐hydroxycinnamate (4‐CIN (Sigma, St Louis, MO)) in 1% methanol‐PBS, or with similar volumes of PBS. ..

    Mouse Assay:

    Article Title: Neural Stem Cells in the Adult Subventricular Zone Oxidize Fatty Acids to Produce Energy and Support Neurogenic Activity
    Article Snippet: .. Pharmacological Inhibition of Fatty Acid and Lactate Transport In Vivo Female C57B/6 mice between 5 and 7 weeks of age were injected intraperitoneally with 40 mg/kg etomoxir (Sigma, St Louis, MO) in PBS, 40 mg/kg alpha‐cyano‐4‐hydroxycinnamate (4‐CIN (Sigma, St Louis, MO)) in 1% methanol‐PBS, or with similar volumes of PBS. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore cnqx
    Cytotoxic <t>NMDA</t> levels abrogate the differential effect of FE65 protein deficiencies on NMDAR-activation dependent APP processing (a) WT and FE65/FE65L1 DKO cortical neurons (DIV 13) were exposed to NMDA (100 μM) for the indicated times. MK801, an NMDA antagonist, was co-incubated with NMDA for the 2.5 hr treatment group. (b) WT or DKO neurons (DIV13) were exposed to the indicated doses of NMDA for 3 hr. (c) WT neurons (DIV 13) were exposed to NMDA (100 μM) or AMPA (10 μM) alone, or in combination with MK801 (10 μM) or <t>CNQX</t> (10 μM) for 3 hr. APP695, APP-CTFs and HMW species were detected by Western blot analysis using the APP C-terminal antibody (A8717). p-APP, pThr668-APP was detected on the same blot with a pThr668APP specific antibody and β-actin was used as a loading control.
    Cnqx, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cnqx/product/Millipore
    Average 99 stars, based on 46 article reviews
    Price from $9.99 to $1999.99
    cnqx - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    93
    Millipore 4 cyano 3 methylisoquinoline
    Ectopically expressed recombinant TRAP220/Med1 is phosphorylated in vivo and in vitro by ERK. (A and B) HeLa cells (8 × 10 5 ) were transiently transfected with pSG5-HA-TRAP220 (4 μg) alone (A) or together with either HA-MKK1(NΔ4) (4 μg) or constitutively (constit.) active ERK2 (4 μg) (B). Twelve hours posttransfection, cells were serum starved for 24 h and then incubated in phosphate-free media containing 32 P i (0.2 mCi/ml) with or without EGF (100 ng/ml) or U0126 (30 μM) for 30 min. Anti-HA immunoprecipitates were resolved by 8% SDS-PAGE and exposed for autoradiography. (C) As a control for expression, HeLa cells were transfected with pSG5-HA-TRAP220 in the absence of 32 P and probed by immunoblotting with anti-HA antibodies. (D) HeLa cells were transiently transfected with pSG5-HA-TRAP220 as in panel A. Twelve hours posttransfection, cells were serum starved for 24 h and then pretreated with the kinase inhibitors U0126 (30 μM), SB202190 (2.5 μM), L-JNK-1 (1 μM), <t>4-cyano-3-methylisoquinoline</t> (300 nM), and bisindolymaleimide I (Bisindolyl.; 100 nM) for 90 min as indicated. The cells were then incubated in fresh phosphate-free media containing 32 P i (0.2 mCi/ml) in the presence or absence of EGF (100 ng/ml) for 20 min. Anti-HA immunoprecipitates were resolved by SDS-PAGE and exposed for autoradiography. (E) MAPK-ERK phosphorylates TRAP220/Med1 in vitro. Purified baculovirally expressed full-length HA-TRAP220/Med1 (50 ng) was incubated with [γ- 32 P]ATP in the presence or absence of purified ERK1 (1 ng) as indicated. Reactions were resolved by SDS-PAGE and exposed for autoradiography (top panel). As a loading control, parallel experiments were performed in the absence of [ 32 P]ATP, resolved by 8% SDS-PAGE, and probed by immunoblotting with anti-HA antibody (bottom panel).
    4 Cyano 3 Methylisoquinoline, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/4 cyano 3 methylisoquinoline/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    4 cyano 3 methylisoquinoline - by Bioz Stars, 2020-11
    93/100 stars
      Buy from Supplier

    99
    Millipore tyrphostin ag 490
    <t>Tyrphostin</t> <t>AG</t> 490 treatment increased expression of PPARγ in SGBS
    Tyrphostin Ag 490, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tyrphostin ag 490/product/Millipore
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    tyrphostin ag 490 - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    Image Search Results


    Cytotoxic NMDA levels abrogate the differential effect of FE65 protein deficiencies on NMDAR-activation dependent APP processing (a) WT and FE65/FE65L1 DKO cortical neurons (DIV 13) were exposed to NMDA (100 μM) for the indicated times. MK801, an NMDA antagonist, was co-incubated with NMDA for the 2.5 hr treatment group. (b) WT or DKO neurons (DIV13) were exposed to the indicated doses of NMDA for 3 hr. (c) WT neurons (DIV 13) were exposed to NMDA (100 μM) or AMPA (10 μM) alone, or in combination with MK801 (10 μM) or CNQX (10 μM) for 3 hr. APP695, APP-CTFs and HMW species were detected by Western blot analysis using the APP C-terminal antibody (A8717). p-APP, pThr668-APP was detected on the same blot with a pThr668APP specific antibody and β-actin was used as a loading control.

    Journal: Journal of neurochemistry

    Article Title: FE65 Proteins Regulate NMDA Receptor Activation-induced Amyloid Precursor Protein Processing

    doi: 10.1111/j.1471-4159.2011.07419.x

    Figure Lengend Snippet: Cytotoxic NMDA levels abrogate the differential effect of FE65 protein deficiencies on NMDAR-activation dependent APP processing (a) WT and FE65/FE65L1 DKO cortical neurons (DIV 13) were exposed to NMDA (100 μM) for the indicated times. MK801, an NMDA antagonist, was co-incubated with NMDA for the 2.5 hr treatment group. (b) WT or DKO neurons (DIV13) were exposed to the indicated doses of NMDA for 3 hr. (c) WT neurons (DIV 13) were exposed to NMDA (100 μM) or AMPA (10 μM) alone, or in combination with MK801 (10 μM) or CNQX (10 μM) for 3 hr. APP695, APP-CTFs and HMW species were detected by Western blot analysis using the APP C-terminal antibody (A8717). p-APP, pThr668-APP was detected on the same blot with a pThr668APP specific antibody and β-actin was used as a loading control.

    Article Snippet: NMDA, AMPA, MK-801, CNQX (6-cyano-7-nitroquinoxaline-2,3-dione), calpeptin, lactacystin and MG132 were purchased from Calbiochem.

    Techniques: Activation Assay, Incubation, Western Blot

    Ectopically expressed recombinant TRAP220/Med1 is phosphorylated in vivo and in vitro by ERK. (A and B) HeLa cells (8 × 10 5 ) were transiently transfected with pSG5-HA-TRAP220 (4 μg) alone (A) or together with either HA-MKK1(NΔ4) (4 μg) or constitutively (constit.) active ERK2 (4 μg) (B). Twelve hours posttransfection, cells were serum starved for 24 h and then incubated in phosphate-free media containing 32 P i (0.2 mCi/ml) with or without EGF (100 ng/ml) or U0126 (30 μM) for 30 min. Anti-HA immunoprecipitates were resolved by 8% SDS-PAGE and exposed for autoradiography. (C) As a control for expression, HeLa cells were transfected with pSG5-HA-TRAP220 in the absence of 32 P and probed by immunoblotting with anti-HA antibodies. (D) HeLa cells were transiently transfected with pSG5-HA-TRAP220 as in panel A. Twelve hours posttransfection, cells were serum starved for 24 h and then pretreated with the kinase inhibitors U0126 (30 μM), SB202190 (2.5 μM), L-JNK-1 (1 μM), 4-cyano-3-methylisoquinoline (300 nM), and bisindolymaleimide I (Bisindolyl.; 100 nM) for 90 min as indicated. The cells were then incubated in fresh phosphate-free media containing 32 P i (0.2 mCi/ml) in the presence or absence of EGF (100 ng/ml) for 20 min. Anti-HA immunoprecipitates were resolved by SDS-PAGE and exposed for autoradiography. (E) MAPK-ERK phosphorylates TRAP220/Med1 in vitro. Purified baculovirally expressed full-length HA-TRAP220/Med1 (50 ng) was incubated with [γ- 32 P]ATP in the presence or absence of purified ERK1 (1 ng) as indicated. Reactions were resolved by SDS-PAGE and exposed for autoradiography (top panel). As a loading control, parallel experiments were performed in the absence of [ 32 P]ATP, resolved by 8% SDS-PAGE, and probed by immunoblotting with anti-HA antibody (bottom panel).

    Journal: Molecular and Cellular Biology

    Article Title: Activation of TRAP/Mediator Subunit TRAP220/Med1 Is Regulated by Mitogen-Activated Protein Kinase-Dependent Phosphorylation

    doi: 10.1128/MCB.25.24.10695-10710.2005

    Figure Lengend Snippet: Ectopically expressed recombinant TRAP220/Med1 is phosphorylated in vivo and in vitro by ERK. (A and B) HeLa cells (8 × 10 5 ) were transiently transfected with pSG5-HA-TRAP220 (4 μg) alone (A) or together with either HA-MKK1(NΔ4) (4 μg) or constitutively (constit.) active ERK2 (4 μg) (B). Twelve hours posttransfection, cells were serum starved for 24 h and then incubated in phosphate-free media containing 32 P i (0.2 mCi/ml) with or without EGF (100 ng/ml) or U0126 (30 μM) for 30 min. Anti-HA immunoprecipitates were resolved by 8% SDS-PAGE and exposed for autoradiography. (C) As a control for expression, HeLa cells were transfected with pSG5-HA-TRAP220 in the absence of 32 P and probed by immunoblotting with anti-HA antibodies. (D) HeLa cells were transiently transfected with pSG5-HA-TRAP220 as in panel A. Twelve hours posttransfection, cells were serum starved for 24 h and then pretreated with the kinase inhibitors U0126 (30 μM), SB202190 (2.5 μM), L-JNK-1 (1 μM), 4-cyano-3-methylisoquinoline (300 nM), and bisindolymaleimide I (Bisindolyl.; 100 nM) for 90 min as indicated. The cells were then incubated in fresh phosphate-free media containing 32 P i (0.2 mCi/ml) in the presence or absence of EGF (100 ng/ml) for 20 min. Anti-HA immunoprecipitates were resolved by SDS-PAGE and exposed for autoradiography. (E) MAPK-ERK phosphorylates TRAP220/Med1 in vitro. Purified baculovirally expressed full-length HA-TRAP220/Med1 (50 ng) was incubated with [γ- 32 P]ATP in the presence or absence of purified ERK1 (1 ng) as indicated. Reactions were resolved by SDS-PAGE and exposed for autoradiography (top panel). As a loading control, parallel experiments were performed in the absence of [ 32 P]ATP, resolved by 8% SDS-PAGE, and probed by immunoblotting with anti-HA antibody (bottom panel).

    Article Snippet: The kinase inhibitors U0126, SB202190, roscovitine, 4-cyano-3-methylisoquinoline and bisindolylmaleimide were all from Calbiochem.

    Techniques: Recombinant, In Vivo, In Vitro, Transfection, Incubation, SDS Page, Autoradiography, Expressing, Purification

    Tyrphostin AG 490 treatment increased expression of PPARγ in SGBS

    Journal: Immunogenetics

    Article Title: Influence of Tyrphostin AG490 on the expression of diabetes associated markers in human dipocytes

    doi: 10.1007/s00251-012-0659-4

    Figure Lengend Snippet: Tyrphostin AG 490 treatment increased expression of PPARγ in SGBS

    Article Snippet: Tyrphostin AG 490 (EMD Chemicals, Gibbstown, NJ), 5 mg, was initially dissolved in sterile DMSO (EMD Chemicals) and brought to final volume by sterile PBS.

    Techniques: Expressing

    The effects of rosiglitazone and Tyrphostin AG 490 on phosphorylation of STAT3 and STAT5 in SGBS

    Journal: Immunogenetics

    Article Title: Influence of Tyrphostin AG490 on the expression of diabetes associated markers in human dipocytes

    doi: 10.1007/s00251-012-0659-4

    Figure Lengend Snippet: The effects of rosiglitazone and Tyrphostin AG 490 on phosphorylation of STAT3 and STAT5 in SGBS

    Article Snippet: Tyrphostin AG 490 (EMD Chemicals, Gibbstown, NJ), 5 mg, was initially dissolved in sterile DMSO (EMD Chemicals) and brought to final volume by sterile PBS.

    Techniques:

    Administration of Tyrphostin AG 490 reduces blood glucose level in pre-diabetic NOD mice

    Journal: Immunogenetics

    Article Title: Influence of Tyrphostin AG490 on the expression of diabetes associated markers in human dipocytes

    doi: 10.1007/s00251-012-0659-4

    Figure Lengend Snippet: Administration of Tyrphostin AG 490 reduces blood glucose level in pre-diabetic NOD mice

    Article Snippet: Tyrphostin AG 490 (EMD Chemicals, Gibbstown, NJ), 5 mg, was initially dissolved in sterile DMSO (EMD Chemicals) and brought to final volume by sterile PBS.

    Techniques: Mouse Assay

    Inhibition of signal transducer and activator of transcription 3 (STAT3) by AG-490 and its consequences on apoptosis signaling in hepatocytes. ( A ) Hepatitis B virus (HBV)–infected differentiated HepaRG cells were either treated with 0.1% dimethyl sulfoxide (DMSO) or with different amounts of AG-490 (0.2 μM, 1 μM, 5 μM, 20 μM, and 100 μM); 24 hours after administration of AG-490, cell viability was analyzed by CellTiter-Blue Cell Viability Assay. Values are given as median ± SD (n = 3; statistical significance relative to DMSO-control; Student's t test). ( B ) Mock- or HBV-infected primary human hepatocytes (PHHs) were either incubated with 0.1% DMSO or with 100 μM of AG-490 for 24 hours. Nuclear or cytosolic proteins were prepared using the NE-PER Nuclear and Cytoplasmic extraction Reagents. Inhibition of STAT3 phosphorylation (upper panel) was analyzed by Western blotting using nuclear proteins and anti-pSTAT3 antibodies. Purity of nuclear protein preparations was controlled by Western blotting using antialbumin antibodies. Cleavage of poly(ADP-ribose) polymerase (PARP) (lower panel) was analyzed by Western blotting using nuclear proteins and anti–poly(ADP-ribose) polymerase (PARP) antibodies. Positions of 116 kDa uncleaved (upper band) and 85 kDa cleaved PARP (lower band) are indicated. Membranes were reprobed with anti-lamin antibodies to control equal protein loading. ( C ) Caspase 3/7 activity in mock- or HBV-infected PHHs treated with either DMSO or with 100 μM AG-490 was measured by luminescent caspase 3/7 assay. Mean ± SD is given (n = 3; *** P

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Hepatitis B Virus Activates Signal Transducer and Activator of Transcription 3 Supporting Hepatocyte Survival and Virus Replication

    doi: 10.1016/j.jcmgh.2017.07.003

    Figure Lengend Snippet: Inhibition of signal transducer and activator of transcription 3 (STAT3) by AG-490 and its consequences on apoptosis signaling in hepatocytes. ( A ) Hepatitis B virus (HBV)–infected differentiated HepaRG cells were either treated with 0.1% dimethyl sulfoxide (DMSO) or with different amounts of AG-490 (0.2 μM, 1 μM, 5 μM, 20 μM, and 100 μM); 24 hours after administration of AG-490, cell viability was analyzed by CellTiter-Blue Cell Viability Assay. Values are given as median ± SD (n = 3; statistical significance relative to DMSO-control; Student's t test). ( B ) Mock- or HBV-infected primary human hepatocytes (PHHs) were either incubated with 0.1% DMSO or with 100 μM of AG-490 for 24 hours. Nuclear or cytosolic proteins were prepared using the NE-PER Nuclear and Cytoplasmic extraction Reagents. Inhibition of STAT3 phosphorylation (upper panel) was analyzed by Western blotting using nuclear proteins and anti-pSTAT3 antibodies. Purity of nuclear protein preparations was controlled by Western blotting using antialbumin antibodies. Cleavage of poly(ADP-ribose) polymerase (PARP) (lower panel) was analyzed by Western blotting using nuclear proteins and anti–poly(ADP-ribose) polymerase (PARP) antibodies. Positions of 116 kDa uncleaved (upper band) and 85 kDa cleaved PARP (lower band) are indicated. Membranes were reprobed with anti-lamin antibodies to control equal protein loading. ( C ) Caspase 3/7 activity in mock- or HBV-infected PHHs treated with either DMSO or with 100 μM AG-490 was measured by luminescent caspase 3/7 assay. Mean ± SD is given (n = 3; *** P

    Article Snippet: Chemicals The pharmacological inhibitor AG-490 was purchased from Calbiochem (San Diego, CA).

    Techniques: Inhibition, Infection, Viability Assay, Incubation, Western Blot, Activity Assay

    Inhibition of signal transducer and activator of transcription 3 by AG-490 and its consequences for hepatitis B virus (HBV) infection. HBV-infected PHHs ( A, B ) or HepaRG cells ( C, D ) were treated for 24 hours either with dimethyl sulfoxide (DMSO) or with AG-490 (20 or 100 μM). ( A, C ) Expression levels of HBV pregenomic RNA (pgRNA) and HNF4α, HNF3α, and HNF3β genes were determined by real-time reverse transcription polymerase chain reaction. Gene expression levels in HBV-infected cells treated with DMSO were set to 100%. ( B, D ) Levels of hepatitis B early antigen (HBeAg) secreted into the cell culture medium were determined by as signal-to-control (S/CO) ratio by enzyme-linked immunosorbent assay at 24 hours after treatment with AG-490. All values are shown as mean ± SD, statistical significance was calculated relative to the respective DMSO-control sample (n = 3; * P

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: Hepatitis B Virus Activates Signal Transducer and Activator of Transcription 3 Supporting Hepatocyte Survival and Virus Replication

    doi: 10.1016/j.jcmgh.2017.07.003

    Figure Lengend Snippet: Inhibition of signal transducer and activator of transcription 3 by AG-490 and its consequences for hepatitis B virus (HBV) infection. HBV-infected PHHs ( A, B ) or HepaRG cells ( C, D ) were treated for 24 hours either with dimethyl sulfoxide (DMSO) or with AG-490 (20 or 100 μM). ( A, C ) Expression levels of HBV pregenomic RNA (pgRNA) and HNF4α, HNF3α, and HNF3β genes were determined by real-time reverse transcription polymerase chain reaction. Gene expression levels in HBV-infected cells treated with DMSO were set to 100%. ( B, D ) Levels of hepatitis B early antigen (HBeAg) secreted into the cell culture medium were determined by as signal-to-control (S/CO) ratio by enzyme-linked immunosorbent assay at 24 hours after treatment with AG-490. All values are shown as mean ± SD, statistical significance was calculated relative to the respective DMSO-control sample (n = 3; * P

    Article Snippet: Chemicals The pharmacological inhibitor AG-490 was purchased from Calbiochem (San Diego, CA).

    Techniques: Inhibition, Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Enzyme-linked Immunosorbent Assay