dentilisin activity  (Millipore)


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  • 85
    Name:
    2 6 Di tert butyl d9 4 methyl phenol 3 5 O d3
    Description:

    Catalog Number:
    452505
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    Structured Review

    Millipore dentilisin activity
    2 6 Di tert butyl d9 4 methyl phenol 3 5 O d3

    https://www.bioz.com/result/dentilisin activity/product/Millipore
    Average 85 stars, based on 6182 article reviews
    Price from $9.99 to $1999.99
    dentilisin activity - by Bioz Stars, 2020-08
    85/100 stars

    Images

    1) Product Images from "Sequence divergence in the Treponema denticola FhbB protein and its impact on factor H binding"

    Article Title: Sequence divergence in the Treponema denticola FhbB protein and its impact on factor H binding

    Journal: Molecular oral microbiology

    doi: 10.1111/omi.12027

    Strain 33521 lacks dentilisin activity and fails to degrade bound FH. (A) Dentilisin activity of strain 33521 was analyzed by measuring SAAPFNA cleavage. Strain 35405 and 35405-CCE (a dentilisin deficient strain) served as positive and negative controls, respectively. All methods were as detailed in the text. (B) The ability of each strain to cleave FH was assessed through immunoblot analysis of FH that had been incubated with whole cells (as indicated). FH breakdown was monitored using anti-human FH antiserum.
    Figure Legend Snippet: Strain 33521 lacks dentilisin activity and fails to degrade bound FH. (A) Dentilisin activity of strain 33521 was analyzed by measuring SAAPFNA cleavage. Strain 35405 and 35405-CCE (a dentilisin deficient strain) served as positive and negative controls, respectively. All methods were as detailed in the text. (B) The ability of each strain to cleave FH was assessed through immunoblot analysis of FH that had been incubated with whole cells (as indicated). FH breakdown was monitored using anti-human FH antiserum.

    Techniques Used: Activity Assay, Incubation

    2) Product Images from "Cornus officinalis Methanol Extract Upregulates Melanogenesis in Melan-a Cells"

    Article Title: Cornus officinalis Methanol Extract Upregulates Melanogenesis in Melan-a Cells

    Journal: Toxicological Research

    doi: 10.5487/TR.2015.31.2.165

    Electron-donating capabilities of 2,6-di-tert-butylate hydroxytoluene (BHT) and Cornus officinalis methanol extract (COME). Values are means ± SD of three measurements.
    Figure Legend Snippet: Electron-donating capabilities of 2,6-di-tert-butylate hydroxytoluene (BHT) and Cornus officinalis methanol extract (COME). Values are means ± SD of three measurements.

    Techniques Used:

    3) Product Images from "Melanogenesis inhibitory effect of aerial part of Pueraria thunbergiana in vitro and in vivo"

    Article Title: Melanogenesis inhibitory effect of aerial part of Pueraria thunbergiana in vitro and in vivo

    Journal: Archives of Dermatological Research

    doi: 10.1007/s00403-014-1489-z

    Effects of aerial part of P. thunbergiana on regulation of Akt/GSK-3β signal. Melanin contents were evaluated. Cells were pretreated in the absence (−) or presence (+) of LY294002 (20 µM) for 1 h and then cultured without (−) or with (+) 50 µg/mL of No. 6 ( a ) and 7 ( b ) for 48 h. *** p
    Figure Legend Snippet: Effects of aerial part of P. thunbergiana on regulation of Akt/GSK-3β signal. Melanin contents were evaluated. Cells were pretreated in the absence (−) or presence (+) of LY294002 (20 µM) for 1 h and then cultured without (−) or with (+) 50 µg/mL of No. 6 ( a ) and 7 ( b ) for 48 h. *** p

    Techniques Used: Cell Culture

    4) Product Images from "Plasma sample based analysis of gastric cancer progression using targeted metabolomics"

    Article Title: Plasma sample based analysis of gastric cancer progression using targeted metabolomics

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-17921-x

    Typical chromatograms of the Trp and Phe metabolites extracted from the analysis of spiked plasma sample. 1: 3-indoleacetonitrile; 2: quinolinic acid; 3: aminophenol; 4: 3-hydroxykynurenine; 5: p-tyrosine; 6: m-tyrosine; 7: serotonin; 8: 5-hydroxytryptophan; 9: o-tyrosine; 10: kynurenine; 11: phenylalanine; 12: N-formylkynurenine; 13: hydroxyanthranillic acid; 14: tryptophan; 15: xanthurenic acid; 16: tryptamine; 17: kynurenic acid; 18: 5-methoxytryptamine; 19: 4-chlorokynurenine; 20: N-acetylserotonin; 21: phenylacetylglutamine; 22: 6-hydroxymelatonin; 23: indole-3-acetamide; 24: anthranillic acid; 25: formyl-acetylmethoxykynurenamine; 26: indolelactic acid; 27: melatonin; 28: 3-indoleacetic acid; 29: tryptophol; 30: serotonin-D 4 ; 31: 5-hydroxytryptophan-D 4 ; 32: kynurenine-D 4 ; 33: phenylalanine-D 5 ; 34: tryptophan-D 5 ; 35: xanthurenic acid-D 4 ; 36: kynurenic acid-D 5 ; 37: tryptamine-D 4 ; 38: 4-chloro-kynurenine- 13 C 2 , 15 N; 39: phenylacetylglutamine-D 5 ; 40: 6-hydroxymelatonin-D 4 ; 41: indole-3-acetamide-D 5 ; 42: melatonin-D 4 .
    Figure Legend Snippet: Typical chromatograms of the Trp and Phe metabolites extracted from the analysis of spiked plasma sample. 1: 3-indoleacetonitrile; 2: quinolinic acid; 3: aminophenol; 4: 3-hydroxykynurenine; 5: p-tyrosine; 6: m-tyrosine; 7: serotonin; 8: 5-hydroxytryptophan; 9: o-tyrosine; 10: kynurenine; 11: phenylalanine; 12: N-formylkynurenine; 13: hydroxyanthranillic acid; 14: tryptophan; 15: xanthurenic acid; 16: tryptamine; 17: kynurenic acid; 18: 5-methoxytryptamine; 19: 4-chlorokynurenine; 20: N-acetylserotonin; 21: phenylacetylglutamine; 22: 6-hydroxymelatonin; 23: indole-3-acetamide; 24: anthranillic acid; 25: formyl-acetylmethoxykynurenamine; 26: indolelactic acid; 27: melatonin; 28: 3-indoleacetic acid; 29: tryptophol; 30: serotonin-D 4 ; 31: 5-hydroxytryptophan-D 4 ; 32: kynurenine-D 4 ; 33: phenylalanine-D 5 ; 34: tryptophan-D 5 ; 35: xanthurenic acid-D 4 ; 36: kynurenic acid-D 5 ; 37: tryptamine-D 4 ; 38: 4-chloro-kynurenine- 13 C 2 , 15 N; 39: phenylacetylglutamine-D 5 ; 40: 6-hydroxymelatonin-D 4 ; 41: indole-3-acetamide-D 5 ; 42: melatonin-D 4 .

    Techniques Used:

    5) Product Images from "Game-changing restraint of Ros-damaged phenylalanine, upon tumor metastasis"

    Article Title: Game-changing restraint of Ros-damaged phenylalanine, upon tumor metastasis

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-017-0147-8

    m -Tyr inhibits spontaneous metastasis a Schematic representation of the in vivo assay carried out to assess the role of m-Tyr in spontaneous metastasis using three metastatic murine mammary carcinomas C7HI, LMM3, and 4T1. For C7HI tumors, BALB/c mice were s.c. injected with 2 × 10 5 cells and 40 days later animals received a daily i.v. injection of m -Tyr (67 mg/kg/day) or saline for the following 20 consecutive days. A third group was killed at day 40 to evaluate the number of metastases at the onset of treatment (basal). At day 60, all treated and control mice were killed and metastases counted. For LMM3 tumors and 4T1 tumors, BALB/c mice were s.c. injected with 2 × 10 5 cells and 25 days later animals received a daily i.v. injection of m -Tyr or saline as previously described. A third group was killed at day 25 to evaluate the number of metastases at the onset of treatment (basal). At day 45, all treated and control mice were EUTHANIZE and metastases counted. All experiments were done in triplicates. b Representative H E staining of pulmonary and draining lymph nodes metastasis of mice bearing s.c. LMM3 tumors (×25, Scale bar = 100 μm). Arrows point sites of tumor cells in the same field. c Pulmonary C7HI, LMM3, or 4T1 metastases for m -Tyr treatment. d Schematic representation of the in vivo assay carried out to assess the role of m -Tyr effect on the survival of LMM3-excised mice exhibiting established lung metastases at the time of surgery. BALB/c mice were s.c. inoculated with 2 × 10 5 LMM3 tumor cells. Twenty-five days later, tumor-bearing mice were surgically operated to remove the tumor and the remaining mice were killed to evaluate the number of lung metastases at the time of surgery. Tumor-excised mice were divided into two groups: m -Tyr (daily i.v. injection of m -Tyr, 67 mg/kg, for consecutive 35 days) and control group (daily i.v. injection of saline for consecutive 35 days). e Kaplan–Meier estimator for survival indicating the percentage of survivors for m -Tyr-treated vs. control mice as a function of the days after tumor excision. * P
    Figure Legend Snippet: m -Tyr inhibits spontaneous metastasis a Schematic representation of the in vivo assay carried out to assess the role of m-Tyr in spontaneous metastasis using three metastatic murine mammary carcinomas C7HI, LMM3, and 4T1. For C7HI tumors, BALB/c mice were s.c. injected with 2 × 10 5 cells and 40 days later animals received a daily i.v. injection of m -Tyr (67 mg/kg/day) or saline for the following 20 consecutive days. A third group was killed at day 40 to evaluate the number of metastases at the onset of treatment (basal). At day 60, all treated and control mice were killed and metastases counted. For LMM3 tumors and 4T1 tumors, BALB/c mice were s.c. injected with 2 × 10 5 cells and 25 days later animals received a daily i.v. injection of m -Tyr or saline as previously described. A third group was killed at day 25 to evaluate the number of metastases at the onset of treatment (basal). At day 45, all treated and control mice were EUTHANIZE and metastases counted. All experiments were done in triplicates. b Representative H E staining of pulmonary and draining lymph nodes metastasis of mice bearing s.c. LMM3 tumors (×25, Scale bar = 100 μm). Arrows point sites of tumor cells in the same field. c Pulmonary C7HI, LMM3, or 4T1 metastases for m -Tyr treatment. d Schematic representation of the in vivo assay carried out to assess the role of m -Tyr effect on the survival of LMM3-excised mice exhibiting established lung metastases at the time of surgery. BALB/c mice were s.c. inoculated with 2 × 10 5 LMM3 tumor cells. Twenty-five days later, tumor-bearing mice were surgically operated to remove the tumor and the remaining mice were killed to evaluate the number of lung metastases at the time of surgery. Tumor-excised mice were divided into two groups: m -Tyr (daily i.v. injection of m -Tyr, 67 mg/kg, for consecutive 35 days) and control group (daily i.v. injection of saline for consecutive 35 days). e Kaplan–Meier estimator for survival indicating the percentage of survivors for m -Tyr-treated vs. control mice as a function of the days after tumor excision. * P

    Techniques Used: In Vivo, Mouse Assay, Injection, Staining

    6) Product Images from "Multicomponent molecular memory"

    Article Title: Multicomponent molecular memory

    Journal: Nature Communications

    doi: 10.1038/s41467-020-14455-1

    Using sparse mixture mapping and multi-peak readout to improve data recovery. a Every 16 bits of data is mapped onto a sparse mixture, based on a 512-Ugi library subset, in which only 32 of the 512 of the compounds are present. There are ~ \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${2}^{169}$$\end{document} 2 169 such mixtures, but only \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${2}^{16}$$\end{document} 2 16 are mapped to data. b Simulated read error rates, before and after decoding. Experimental results from single peak ( c ) and multi-peak ( e ) detection are also shown. c A 24,336 pixel binary image of angels at a dinner table from a 16th century print 40 , recovered with single (sodiated) peak readout (96.67% accurate). d Histograms of the raw compound errors per mixture for each recovery scheme. e The digital image recovered with multi-peak detection (99.89% accurate).
    Figure Legend Snippet: Using sparse mixture mapping and multi-peak readout to improve data recovery. a Every 16 bits of data is mapped onto a sparse mixture, based on a 512-Ugi library subset, in which only 32 of the 512 of the compounds are present. There are ~ \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${2}^{169}$$\end{document} 2 169 such mixtures, but only \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${2}^{16}$$\end{document} 2 16 are mapped to data. b Simulated read error rates, before and after decoding. Experimental results from single peak ( c ) and multi-peak ( e ) detection are also shown. c A 24,336 pixel binary image of angels at a dinner table from a 16th century print 40 , recovered with single (sodiated) peak readout (96.67% accurate). d Histograms of the raw compound errors per mixture for each recovery scheme. e The digital image recovered with multi-peak detection (99.89% accurate).

    Techniques Used:

    Library analysis. a A color-coded map of the signal-to-noise ratio (SNR) of sodiated product peaks across a 1500-Ugi library. b A histogram of the 50 strongest peaks in each product spectra, offset by their expected Ugi product masses (M). Several common salt adducts and isotopes are labeled. Inset: A histogram of the monoisotoptic masses of the 1500-compound library. c The true positive rate (TPR) and false positive rate (FPR) as a function of the SNR threshold for the M+Na peaks. The dashed line (SNR \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\approx$$\end{document} ≈ 31) jointly optimizes the true and false positive rates ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\,\text{TPR}\,=89.73 \%$$\end{document} TPR = 89.73 % , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\,\text{FPR}\,=8.71 \%$$\end{document} FPR = 8.71 % ).
    Figure Legend Snippet: Library analysis. a A color-coded map of the signal-to-noise ratio (SNR) of sodiated product peaks across a 1500-Ugi library. b A histogram of the 50 strongest peaks in each product spectra, offset by their expected Ugi product masses (M). Several common salt adducts and isotopes are labeled. Inset: A histogram of the monoisotoptic masses of the 1500-compound library. c The true positive rate (TPR) and false positive rate (FPR) as a function of the SNR threshold for the M+Na peaks. The dashed line (SNR \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\approx$$\end{document} ≈ 31) jointly optimizes the true and false positive rates ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\,\text{TPR}\,=89.73 \%$$\end{document} TPR = 89.73 % , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\,\text{FPR}\,=8.71 \%$$\end{document} FPR = 8.71 % ).

    Techniques Used: Labeling

    7) Product Images from "New insights into the metabolism of organomercury compounds: Mercury-containing cysteine S-conjugates are substrates of human glutamine transaminase K and potent inactivators of cystathionine ?-lyase"

    Article Title: New insights into the metabolism of organomercury compounds: Mercury-containing cysteine S-conjugates are substrates of human glutamine transaminase K and potent inactivators of cystathionine ?-lyase

    Journal: Archives of biochemistry and biophysics

    doi: 10.1016/j.abb.2011.11.002

    Effect of HgCl 2 and CH 3 HgCl on the rhGTK-catalyzed transamination of L-phenylalanine with KMB. The reaction mixture (50 μl) contained 20 mM L-phenylalanine, 5 mM KMB, 2.6 mU of enzyme, 100 mM potassium phosphate buffer (pH 7.4) in the presence
    Figure Legend Snippet: Effect of HgCl 2 and CH 3 HgCl on the rhGTK-catalyzed transamination of L-phenylalanine with KMB. The reaction mixture (50 μl) contained 20 mM L-phenylalanine, 5 mM KMB, 2.6 mU of enzyme, 100 mM potassium phosphate buffer (pH 7.4) in the presence

    Techniques Used:

    8) Product Images from "Poly-?-Glutamic Acid Nanoparticles and Aluminum Adjuvant Used as an Adjuvant with a Single Dose of Japanese Encephalitis Virus-Like Particles Provide Effective Protection from Japanese Encephalitis Virus"

    Article Title: Poly-?-Glutamic Acid Nanoparticles and Aluminum Adjuvant Used as an Adjuvant with a Single Dose of Japanese Encephalitis Virus-Like Particles Provide Effective Protection from Japanese Encephalitis Virus

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.05412-11

    Effect of alum and γ-PGA-NPs on the length of protection from a single JE-VLP immunization. Groups of mice (10 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs, JE-VLP, a mixture of JE-VLP
    Figure Legend Snippet: Effect of alum and γ-PGA-NPs on the length of protection from a single JE-VLP immunization. Groups of mice (10 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs, JE-VLP, a mixture of JE-VLP

    Techniques Used: Mouse Assay, Injection

    Effect of alum or γ-PGA-NPs adjuvants on JE-VLP protection against death from JEV infection. Groups of mice were immunized by intraperitoneal injection of 400 μl PBS (black circles; n = 31), 400 μl PBS containing 100 μg
    Figure Legend Snippet: Effect of alum or γ-PGA-NPs adjuvants on JE-VLP protection against death from JEV infection. Groups of mice were immunized by intraperitoneal injection of 400 μl PBS (black circles; n = 31), 400 μl PBS containing 100 μg

    Techniques Used: Infection, Mouse Assay, Injection

    Anti-JEV neutralizing antibody response from JE-VLP with alum or γ-PGA-NPs. Groups of mice (5 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs, JE-VLP, JE-VLP and alum, JE-VLP and γ-PGA-NPs,
    Figure Legend Snippet: Anti-JEV neutralizing antibody response from JE-VLP with alum or γ-PGA-NPs. Groups of mice (5 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs, JE-VLP, JE-VLP and alum, JE-VLP and γ-PGA-NPs,

    Techniques Used: Mouse Assay, Injection

    Effect of alum and γ-PGA-NPs on the continuous secretion of anti-JEV neutralizing antibodies following a single JE-VLP immunization. (A) Groups of mice (5 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs,
    Figure Legend Snippet: Effect of alum and γ-PGA-NPs on the continuous secretion of anti-JEV neutralizing antibodies following a single JE-VLP immunization. (A) Groups of mice (5 per group) were given a single intraperitoneal injection of PBS, PBS containing alum, γ-PGA-NPs,

    Techniques Used: Mouse Assay, Injection

    9) Product Images from "Molecular Mechanism of Activation of Human Cdc7 Kinase"

    Article Title: Molecular Mechanism of Activation of Human Cdc7 Kinase

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M111.243311

    Mapping of the segments of Cdc7 required for interaction with ASK. A , summary of various deletion derivatives of human Cdc7 cloned on monomeric Kusabira Orange2-HA fusion vector or on pME18S-HA. Black boxes represent the deleted segments in each mutant.
    Figure Legend Snippet: Mapping of the segments of Cdc7 required for interaction with ASK. A , summary of various deletion derivatives of human Cdc7 cloned on monomeric Kusabira Orange2-HA fusion vector or on pME18S-HA. Black boxes represent the deleted segments in each mutant.

    Techniques Used: Clone Assay, Plasmid Preparation, Mutagenesis

    Interaction of an isolated Cdc7 C-terminal polypeptide with ASK. A plasmid expressing FLAG- or Myc-tagged Cdc7 C-terminal polypeptide (432–574) was transfected into 293T cells together with Myc- or GFP-tagged ASK plasmid, respectively, as indicated
    Figure Legend Snippet: Interaction of an isolated Cdc7 C-terminal polypeptide with ASK. A plasmid expressing FLAG- or Myc-tagged Cdc7 C-terminal polypeptide (432–574) was transfected into 293T cells together with Myc- or GFP-tagged ASK plasmid, respectively, as indicated

    Techniques Used: Isolation, Plasmid Preparation, Expressing, Transfection

    Identification of the residues on Cdc7 directly interacting with ASK. A , UV photocross-linking between selected Cdc7-Bpa and ASK. Protein expression was induced in the E. coli ) and Bpa-substituted pETDuet-Cdc7-ASK
    Figure Legend Snippet: Identification of the residues on Cdc7 directly interacting with ASK. A , UV photocross-linking between selected Cdc7-Bpa and ASK. Protein expression was induced in the E. coli ) and Bpa-substituted pETDuet-Cdc7-ASK

    Techniques Used: Expressing

    Characterization of purified Cdc7 kinase (T1, full-length) protein with or without associated ASK subunit. A
    Figure Legend Snippet: Characterization of purified Cdc7 kinase (T1, full-length) protein with or without associated ASK subunit. A

    Techniques Used: Purification

    Two motifs, DAM-1 and DAM-2, of Cdc7 are required for interaction with ASK and kinase activation. pME18S-HA-Cdc7WT, pME18S-HA-Cdc7(448A457; 10A), or pME18S-HA-Cdc7(566A572; 7A) was transfected into 293T cells with or without pME18S-FLAG-ASK, as indicated.
    Figure Legend Snippet: Two motifs, DAM-1 and DAM-2, of Cdc7 are required for interaction with ASK and kinase activation. pME18S-HA-Cdc7WT, pME18S-HA-Cdc7(448A457; 10A), or pME18S-HA-Cdc7(566A572; 7A) was transfected into 293T cells with or without pME18S-FLAG-ASK, as indicated.

    Techniques Used: Activation Assay, Transfection

    Dissection of human Cdc7 kinase, two segments, DAM-1 and DAM-2, required for interaction with ASK. A , schematic representation of human Cdc7 polypeptide showing two essential motifs (DAM-1 and DAM-2 indicated by the pink boxes ) required for interaction
    Figure Legend Snippet: Dissection of human Cdc7 kinase, two segments, DAM-1 and DAM-2, required for interaction with ASK. A , schematic representation of human Cdc7 polypeptide showing two essential motifs (DAM-1 and DAM-2 indicated by the pink boxes ) required for interaction

    Techniques Used: Dissection

    Characterization of purified minimum Cdc7 kinase (T5) protein with or without associated ASK subunit. A , schematic drawing showing the segments of Cdc7 present in the minimum Cdc7 polypeptide (Cdc7T5; shown on top of the Cdc7 bar). Cdc7T5 was expressed
    Figure Legend Snippet: Characterization of purified minimum Cdc7 kinase (T5) protein with or without associated ASK subunit. A , schematic drawing showing the segments of Cdc7 present in the minimum Cdc7 polypeptide (Cdc7T5; shown on top of the Cdc7 bar). Cdc7T5 was expressed

    Techniques Used: Purification

    Requirement of the C-terminal Tail and a Part of Kinase Insert III of Cdc7 for Interaction with ASK and ASK-mediated Kinase Activation
    Figure Legend Snippet: Requirement of the C-terminal Tail and a Part of Kinase Insert III of Cdc7 for Interaction with ASK and ASK-mediated Kinase Activation

    Techniques Used: Activation Assay

    10) Product Images from "Intestinal alkaline phosphatase promotes gut bacterial growth by reducing the concentration of luminal nucleotide triphosphates"

    Article Title: Intestinal alkaline phosphatase promotes gut bacterial growth by reducing the concentration of luminal nucleotide triphosphates

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    doi: 10.1152/ajpgi.00357.2013

    Gut microbiota live in symbiosis with the host. The brush border enzyme IAP appears to play a central role in regulating the microbiota through a mechanism that involves the dephosphorylation of luminal ATP. ATP (and similarly other nucleotide triphosphates
    Figure Legend Snippet: Gut microbiota live in symbiosis with the host. The brush border enzyme IAP appears to play a central role in regulating the microbiota through a mechanism that involves the dephosphorylation of luminal ATP. ATP (and similarly other nucleotide triphosphates

    Techniques Used: De-Phosphorylation Assay

    11) Product Images from "Midgut serine proteases and alternative host plant utilization in Pieris brassicae L."

    Article Title: Midgut serine proteases and alternative host plant utilization in Pieris brassicae L.

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2015.00095

    Gelatin zymograms of midgut samples from field-collected, fourth instar P. brassicae found feeding on leaves of cauliflower (CF-ctrl), cabbage (CB-ctrl), mustard (MT-ctrl), radish (RD-ctrl), and T. majus (TP-ctrl) in (A) absence and presence of STI (5 mg/ml) . Zymograms of samples (B) incubated with inhibitors TLCK (2 mg/ml) and TPCK (2 mg/ml) are also shown. Asterisk denotes activity zone observed only in midgut samples of larvae fed on T. majus .
    Figure Legend Snippet: Gelatin zymograms of midgut samples from field-collected, fourth instar P. brassicae found feeding on leaves of cauliflower (CF-ctrl), cabbage (CB-ctrl), mustard (MT-ctrl), radish (RD-ctrl), and T. majus (TP-ctrl) in (A) absence and presence of STI (5 mg/ml) . Zymograms of samples (B) incubated with inhibitors TLCK (2 mg/ml) and TPCK (2 mg/ml) are also shown. Asterisk denotes activity zone observed only in midgut samples of larvae fed on T. majus .

    Techniques Used: Incubation, Activity Assay

    12) Product Images from "A Retrospective Analysis of the Cartilage Kunitz Protease Inhibitory Proteins Identifies These as Members of the Inter-α-Trypsin Inhibitor Superfamily with Potential Roles in the Protection of the Articulatory Surface"

    Article Title: A Retrospective Analysis of the Cartilage Kunitz Protease Inhibitory Proteins Identifies These as Members of the Inter-α-Trypsin Inhibitor Superfamily with Potential Roles in the Protection of the Articulatory Surface

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20030497

    Schematic representations and proposed inter relationship between SPI species seen in this study. a. α1 microglobulin-bikunin precursor (58–64 kDa), b. bikunin 30–36 kDa, c. bikunin KPI dimer (12–16 kDa) after removal of the 11.7 kDa CS chain from bikunin, d. bikunin KPI monomers (6–9 kDa) with the N-linked 2–4 kDa glycan oligosaccharide attached to the KPI-1 domain which facilitates its isolation by ConA affinity chromatography and the 6 kDa KPI-2 domain which does contain oligosaccharide substitution.
    Figure Legend Snippet: Schematic representations and proposed inter relationship between SPI species seen in this study. a. α1 microglobulin-bikunin precursor (58–64 kDa), b. bikunin 30–36 kDa, c. bikunin KPI dimer (12–16 kDa) after removal of the 11.7 kDa CS chain from bikunin, d. bikunin KPI monomers (6–9 kDa) with the N-linked 2–4 kDa glycan oligosaccharide attached to the KPI-1 domain which facilitates its isolation by ConA affinity chromatography and the 6 kDa KPI-2 domain which does contain oligosaccharide substitution.

    Techniques Used: Isolation, Affinity Chromatography

    Sequence of the CS chain of bikunin as proposed by Ly et al. 2011 [ 11 ] ( a ) and Lord et al. 2013 [ 10 ] ( b ). The KPI SPI domains are shown in ( b ). and the attached N -glycan chain of KPI-1. The over-sulfated CS-D disaccharide is shown which forms part of the MO-225 epitope embedded in the CS-A chain of bikunin. Arrangement of sulfated, disulfated and non-sulfated regions of the CS chain of bikunin are shown. The arrow shows the Asparagine residue on Kunitz domain 1.
    Figure Legend Snippet: Sequence of the CS chain of bikunin as proposed by Ly et al. 2011 [ 11 ] ( a ) and Lord et al. 2013 [ 10 ] ( b ). The KPI SPI domains are shown in ( b ). and the attached N -glycan chain of KPI-1. The over-sulfated CS-D disaccharide is shown which forms part of the MO-225 epitope embedded in the CS-A chain of bikunin. Arrangement of sulfated, disulfated and non-sulfated regions of the CS chain of bikunin are shown. The arrow shows the Asparagine residue on Kunitz domain 1.

    Techniques Used: Sequencing

    13) Product Images from "Pharmaceutical significance of Leuconostoc mesenteroides KS-TN11 isolated from Nile Tilapia, Oreochromis niloticus"

    Article Title: Pharmaceutical significance of Leuconostoc mesenteroides KS-TN11 isolated from Nile Tilapia, Oreochromis niloticus

    Journal: Saudi Pharmaceutical Journal : SPJ

    doi: 10.1016/j.jsps.2018.02.006

    α-Glucosidase inhibitory activity of L. mesenteroides KS-TN11.
    Figure Legend Snippet: α-Glucosidase inhibitory activity of L. mesenteroides KS-TN11.

    Techniques Used: Activity Assay

    14) Product Images from "Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation"

    Article Title: Rapid induction of p62 and GABARAPL1 upon proteasome inhibition promotes survival before autophagy activation

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.201708168

    p62 knockdown in neuroblastoma cells did not reduce protein degradation or increase levels of Ub conjugates but impaired the buildup of polyubiquitinated and polysumoylated proteins in inclusions. (A) p62 knockdown (sh-p62) did not affect the ability of SH-SY5Y cells to degrade long-lived proteins when treated or not with 1 µM BTZ to completely inhibit the proteasome. n = 6. (B) In M17 cells treated with BTZ, stable knockdown of p62 but not Nbr1 or GABARAPL1 reduced the buildup of Ub or SUMO2/3 conjugates in the pellet that contains inclusions. In the supernatant, there are no SUMO2/3-conjugated proteins, and the bands recognized by the SUMO2/3 antibody (asterisk) are not specific. (C and D) In SH-SY5Y cells treated for 16 h with 100 nM BTZ, knockdown of p62 reduced Ub (C) and SUMO2/3 (D) conjugates in the pellet. (C) Knockdown of Nbr1 and GABARAPL1 also reduced Ub conjugates in the pellet. Molecular masses are given in kilodaltons. Error bars indicate SD.
    Figure Legend Snippet: p62 knockdown in neuroblastoma cells did not reduce protein degradation or increase levels of Ub conjugates but impaired the buildup of polyubiquitinated and polysumoylated proteins in inclusions. (A) p62 knockdown (sh-p62) did not affect the ability of SH-SY5Y cells to degrade long-lived proteins when treated or not with 1 µM BTZ to completely inhibit the proteasome. n = 6. (B) In M17 cells treated with BTZ, stable knockdown of p62 but not Nbr1 or GABARAPL1 reduced the buildup of Ub or SUMO2/3 conjugates in the pellet that contains inclusions. In the supernatant, there are no SUMO2/3-conjugated proteins, and the bands recognized by the SUMO2/3 antibody (asterisk) are not specific. (C and D) In SH-SY5Y cells treated for 16 h with 100 nM BTZ, knockdown of p62 reduced Ub (C) and SUMO2/3 (D) conjugates in the pellet. (C) Knockdown of Nbr1 and GABARAPL1 also reduced Ub conjugates in the pellet. Molecular masses are given in kilodaltons. Error bars indicate SD.

    Techniques Used:

    Prolonged and strong inhibition of the proteasome causes cells to induce the mRNAs for nearly all Atg genes and Ub receptors. (A) When treated with 100 nM BTZ for 20 h, SH-SY5Y cells induced the mRNAs for nearly all Atg genes and Ub receptors but less than did p62 and GABARAPL1 . (B) SH-SY5Y cells induced the mRNAs of most Atg genes when treated with 100 nM BTZ for 20 h (right). Reducing the BTZ concentration to 10 nM (right) or the treatment time to 13 h (left) prevented induction. (C and D) MM1.S cells induced the mRNAs of most Atg genes when treated with 20 or 50 nM BTZ for 19 h, although much less than GABARAPL1 and p62 (C). Reducing the BTZ concentration to 10 nM (C) or the treatment time to 14 h (D) failed to induce most genes. In this figure and Fig. 3 , the exact fold increase is shown for p62 and GABARAPL1 because their mRNAs increase > 10-fold. * , P
    Figure Legend Snippet: Prolonged and strong inhibition of the proteasome causes cells to induce the mRNAs for nearly all Atg genes and Ub receptors. (A) When treated with 100 nM BTZ for 20 h, SH-SY5Y cells induced the mRNAs for nearly all Atg genes and Ub receptors but less than did p62 and GABARAPL1 . (B) SH-SY5Y cells induced the mRNAs of most Atg genes when treated with 100 nM BTZ for 20 h (right). Reducing the BTZ concentration to 10 nM (right) or the treatment time to 13 h (left) prevented induction. (C and D) MM1.S cells induced the mRNAs of most Atg genes when treated with 20 or 50 nM BTZ for 19 h, although much less than GABARAPL1 and p62 (C). Reducing the BTZ concentration to 10 nM (C) or the treatment time to 14 h (D) failed to induce most genes. In this figure and Fig. 3 , the exact fold increase is shown for p62 and GABARAPL1 because their mRNAs increase > 10-fold. * , P

    Techniques Used: Inhibition, Concentration Assay

    Nrf1 and NF-E2 activate p62 transcription in control cells and upon proteasome inhibition. (A) When different p62 promoter segments were fused to luciferase and expressed in M17 cells, the segments (P310 [−310 to 37] and P1068) that contain NFE2-E increased luciferase expression by ∼40% upon BTZ (0.1 µM) treatment for 16 h. However, the segment (P270) lacking NFE2-E showed no effect. n = 4. (B) NF-E2 knockdown by siRNA in HEK293A cells (verified by NF-E2 mRNA measurement in the left panel) reduced p62 mRNA (left) and protein (right) in cells treated or not for 16 h with 0.1 µM BTZ. (C) In M17 cells, knockdown of Nrf1 or NF-E2 , but not Nrf2 , reduced p62 level after BTZ treatment. (D) In HEK293A cells, knockdown of both NF-E2 (by siRNA) and Nrf1 (by stably expressing shRNA) caused additive reduction of p62 mRNA (left) and protein (right) after BTZ treatment. Molecular masses are given in kilodaltons. (E and F) Stable knockdown of Nrf1 with shRNA (E) suppressed p62 mRNA in untreated HAP1 cells or cells treated for 16 h with 100 nM BTZ or CFZ, and in SH-SY5Y cells treated with BTZ at indicated conditions (F). (G–J) Knockdown of Nrf1 in SH-SY5Y (G and H) or HAP1 (I and J) cells suppressed p62 mRNA when treated with 100 nM BTZ for 20 h (G and I) but did not suppress the mRNAs for most other Atg genes and Ub receptors (H and J). * , P
    Figure Legend Snippet: Nrf1 and NF-E2 activate p62 transcription in control cells and upon proteasome inhibition. (A) When different p62 promoter segments were fused to luciferase and expressed in M17 cells, the segments (P310 [−310 to 37] and P1068) that contain NFE2-E increased luciferase expression by ∼40% upon BTZ (0.1 µM) treatment for 16 h. However, the segment (P270) lacking NFE2-E showed no effect. n = 4. (B) NF-E2 knockdown by siRNA in HEK293A cells (verified by NF-E2 mRNA measurement in the left panel) reduced p62 mRNA (left) and protein (right) in cells treated or not for 16 h with 0.1 µM BTZ. (C) In M17 cells, knockdown of Nrf1 or NF-E2 , but not Nrf2 , reduced p62 level after BTZ treatment. (D) In HEK293A cells, knockdown of both NF-E2 (by siRNA) and Nrf1 (by stably expressing shRNA) caused additive reduction of p62 mRNA (left) and protein (right) after BTZ treatment. Molecular masses are given in kilodaltons. (E and F) Stable knockdown of Nrf1 with shRNA (E) suppressed p62 mRNA in untreated HAP1 cells or cells treated for 16 h with 100 nM BTZ or CFZ, and in SH-SY5Y cells treated with BTZ at indicated conditions (F). (G–J) Knockdown of Nrf1 in SH-SY5Y (G and H) or HAP1 (I and J) cells suppressed p62 mRNA when treated with 100 nM BTZ for 20 h (G and I) but did not suppress the mRNAs for most other Atg genes and Ub receptors (H and J). * , P

    Techniques Used: Inhibition, Luciferase, Expressing, Stable Transfection, shRNA

    Upon proteasome inhibition, Ub conjugation is required for the induction of p62 and GABARAPL1 . (A) Inhibiting Ub conjugation with 0.5 µM ML-997 greatly suppressed the induction of p62 and GABARAPL1 mRNAs in SH-SY5Y cells treated for 16 h with 100 nM BTZ. (B–D) In RPMI 8226 cells treated for 12 h with 20 nM BTZ, inhibiting Ub conjugation with 0.5 µM TAK-243 (B) depleted Ub conjugates (C) without causing widespread cell death (measured by MTS assay) and suppressed the induction of p62 and GABARAPL1 (D). * , P
    Figure Legend Snippet: Upon proteasome inhibition, Ub conjugation is required for the induction of p62 and GABARAPL1 . (A) Inhibiting Ub conjugation with 0.5 µM ML-997 greatly suppressed the induction of p62 and GABARAPL1 mRNAs in SH-SY5Y cells treated for 16 h with 100 nM BTZ. (B–D) In RPMI 8226 cells treated for 12 h with 20 nM BTZ, inhibiting Ub conjugation with 0.5 µM TAK-243 (B) depleted Ub conjugates (C) without causing widespread cell death (measured by MTS assay) and suppressed the induction of p62 and GABARAPL1 (D). * , P

    Techniques Used: Inhibition, Conjugation Assay, MTS Assay

    Autophagy deficiency ( Atg5 −/− MEFs) or knockdown of p62 or GABARAPL1 but not Nbr1 increased cell sensitivity to killing by BTZ. (A) Atg5 −/− MEFs lost viability much more than WT cells after BTZ treatment. (B) Confirmation of stable knockdown of p62 , Nbr1 , or GABARAPL1 by shRNA in untreated or BTZ-treated (16 h) SH-SY5Y cells. Molecular masses are given in kilodaltons. (C) SH-SY5Y cells expressing sh-p62 or sh-GABARAPL1 but not sh-Nbr1 lost viability more than WT cells did when treated with BTZ for 36 h. (D) Knockdown of p62 or GABRAPL1 also caused M17 cells to lose viability more than WT cells when treated with BTZ for 40 h. Viability was measured with the MTS assay. *, P
    Figure Legend Snippet: Autophagy deficiency ( Atg5 −/− MEFs) or knockdown of p62 or GABARAPL1 but not Nbr1 increased cell sensitivity to killing by BTZ. (A) Atg5 −/− MEFs lost viability much more than WT cells after BTZ treatment. (B) Confirmation of stable knockdown of p62 , Nbr1 , or GABARAPL1 by shRNA in untreated or BTZ-treated (16 h) SH-SY5Y cells. Molecular masses are given in kilodaltons. (C) SH-SY5Y cells expressing sh-p62 or sh-GABARAPL1 but not sh-Nbr1 lost viability more than WT cells did when treated with BTZ for 36 h. (D) Knockdown of p62 or GABRAPL1 also caused M17 cells to lose viability more than WT cells when treated with BTZ for 40 h. Viability was measured with the MTS assay. *, P

    Techniques Used: shRNA, Expressing, MTS Assay

    Proteasome inhibitor treatment causes rapid induction of p62 and GABARAPL1 but not other Atg genes or Ub receptors. (A and B) SH-SY5Y cells were treated with 10 nM BTZ for 13 h (A) or 1 µM for 4 h (B). The mRNAs of all Atg proteins or Ub receptors were measured in this and other figures by real time RT-PCR. (C) Autophagy was activated only after prolonged treatment with a high concentration of proteasome inhibitor. SH-SY5Y cells were treated with 10 or 100 nM BTZ for 5–24 h. To measure autophagy, the levels of LC3-I and lipidated (autophagosome-bound) LC3-II were measured (upper) by Western blotting, and the LC3-II/I ratio was quantified (lower). *, LC3-II/I ratio in cells treated with 100 nM BTZ is higher than that in untreated cells (P
    Figure Legend Snippet: Proteasome inhibitor treatment causes rapid induction of p62 and GABARAPL1 but not other Atg genes or Ub receptors. (A and B) SH-SY5Y cells were treated with 10 nM BTZ for 13 h (A) or 1 µM for 4 h (B). The mRNAs of all Atg proteins or Ub receptors were measured in this and other figures by real time RT-PCR. (C) Autophagy was activated only after prolonged treatment with a high concentration of proteasome inhibitor. SH-SY5Y cells were treated with 10 or 100 nM BTZ for 5–24 h. To measure autophagy, the levels of LC3-I and lipidated (autophagosome-bound) LC3-II were measured (upper) by Western blotting, and the LC3-II/I ratio was quantified (lower). *, LC3-II/I ratio in cells treated with 100 nM BTZ is higher than that in untreated cells (P

    Techniques Used: Quantitative RT-PCR, Concentration Assay, Western Blot

    FoxO3a, p-eIF2α, ATF4, Nrf2, or TFEB are not responsible for the induction of p62 and GABARAPL1 upon proteasome inhibition. (A) Expression of DN-FoxO3a did not affect the ability of HEK293A cells to induce p62 and GABARAPL1 mRNAs upon treatment with BTZ (10 nM) or MG132 (10 µM) for 16 h. (B and C) Unlike proteasome inhibitors, UPR inducers (tunicamycin and thapsigargin) did not cause a large (more than fivefold) induction of p62 or GABARAPL1 . SH-SY5Y cells were treated for 16 h with BTZ (10 nM), MG132 (10 µM), epoxomicin (Epox; 50 nM), tunicamycin (Tunic; 10 µg/ml), or thapsigargin (Thaps; 300 nM), and mRNAs (B) and proteins (C) were measured. (D) To confirm successful knockdown of Nrf2, WT or stable Nrf2 knockdown (by shRNA) SH-SY5Y cells were treated for 16 h with sulforaphane (SFN; 10 µM) to activate Nrf2, and mRNAs of Nrf2 and NQO1, whose expression requires Nrf2, were measured. (E) sh-Nrf2 cells were not defective in inducing p62 or GABARAPL1 mRNAs upon BTZ treatment (10 nM for 16 h). (F and G) Knockdown of TFEB by siRNA in M17 cells was validated by Western blotting (F), but did not reduce the cells’ ability to rapidly induce p62 and GABARAPL1 mRNAs upon BTZ treatment or to induce LC3B mRNA when treated with 0.1 µM BTZ for 20 h (G). * , P
    Figure Legend Snippet: FoxO3a, p-eIF2α, ATF4, Nrf2, or TFEB are not responsible for the induction of p62 and GABARAPL1 upon proteasome inhibition. (A) Expression of DN-FoxO3a did not affect the ability of HEK293A cells to induce p62 and GABARAPL1 mRNAs upon treatment with BTZ (10 nM) or MG132 (10 µM) for 16 h. (B and C) Unlike proteasome inhibitors, UPR inducers (tunicamycin and thapsigargin) did not cause a large (more than fivefold) induction of p62 or GABARAPL1 . SH-SY5Y cells were treated for 16 h with BTZ (10 nM), MG132 (10 µM), epoxomicin (Epox; 50 nM), tunicamycin (Tunic; 10 µg/ml), or thapsigargin (Thaps; 300 nM), and mRNAs (B) and proteins (C) were measured. (D) To confirm successful knockdown of Nrf2, WT or stable Nrf2 knockdown (by shRNA) SH-SY5Y cells were treated for 16 h with sulforaphane (SFN; 10 µM) to activate Nrf2, and mRNAs of Nrf2 and NQO1, whose expression requires Nrf2, were measured. (E) sh-Nrf2 cells were not defective in inducing p62 or GABARAPL1 mRNAs upon BTZ treatment (10 nM for 16 h). (F and G) Knockdown of TFEB by siRNA in M17 cells was validated by Western blotting (F), but did not reduce the cells’ ability to rapidly induce p62 and GABARAPL1 mRNAs upon BTZ treatment or to induce LC3B mRNA when treated with 0.1 µM BTZ for 20 h (G). * , P

    Techniques Used: Inhibition, Expressing, shRNA, Western Blot

    15) Product Images from "GCN2-dependent phosphorylation of eukaryotic translation initiation factor-2? in Arabidopsis"

    Article Title: GCN2-dependent phosphorylation of eukaryotic translation initiation factor-2? in Arabidopsis

    Journal: Journal of Experimental Botany

    doi: 10.1093/jxb/ern169

    (A) Wild-type Arabidopsis, ecotype Landsberg erecta , and gene trap mutant line GT8359 growing in soil. (B) Arabidopsis seedlings growing on agar plates. The plate on the left is a control while the other plates were treated with herbicide as indicated. In each case wild-type Arabidopsis, ecotype Landsberg erecta, was sown on the top half of the plate while Genetrap mutant line GT8359 was sown on the bottom half, as indicated. (C) As for (B), except that the seedlings were fed with the appropriate amino acids to compensate for the herbicide treatments; these were phenylalanine, tryptophan and tyrosine in the case of Glyphosate, histidine in the case of IRL 1803 and isoleucine, leucine and valine for Chlorsulfuron.
    Figure Legend Snippet: (A) Wild-type Arabidopsis, ecotype Landsberg erecta , and gene trap mutant line GT8359 growing in soil. (B) Arabidopsis seedlings growing on agar plates. The plate on the left is a control while the other plates were treated with herbicide as indicated. In each case wild-type Arabidopsis, ecotype Landsberg erecta, was sown on the top half of the plate while Genetrap mutant line GT8359 was sown on the bottom half, as indicated. (C) As for (B), except that the seedlings were fed with the appropriate amino acids to compensate for the herbicide treatments; these were phenylalanine, tryptophan and tyrosine in the case of Glyphosate, histidine in the case of IRL 1803 and isoleucine, leucine and valine for Chlorsulfuron.

    Techniques Used: Mutagenesis

    16) Product Images from "Relationship between changes in the 7-day urticaria activity score after treatment with omalizumab and the responsiveness of basophils to FcεRI stimulation in patients with chronic spontaneous urticaria"

    Article Title: Relationship between changes in the 7-day urticaria activity score after treatment with omalizumab and the responsiveness of basophils to FcεRI stimulation in patients with chronic spontaneous urticaria

    Journal: Asia Pacific Allergy

    doi: 10.5415/apallergy.2020.10.e12

    Comparison of the peripheral blood basophil counts (A, n = 8; B, n = 11), proportion (%) of CD203 high basophils after FcεRI aggregation (C, n = 7; D, n = 7), and proportion (%) of CD203 high basophils following N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) stimulation (E, n = 4 ; F, n = 6) in the BAT-negative and BAT-positive patients before (day 0) and after treatment with omalizumab (day 84). Statistical analyses were performed using the Mann-Whitney U test. BAT, basophil activation test.
    Figure Legend Snippet: Comparison of the peripheral blood basophil counts (A, n = 8; B, n = 11), proportion (%) of CD203 high basophils after FcεRI aggregation (C, n = 7; D, n = 7), and proportion (%) of CD203 high basophils following N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) stimulation (E, n = 4 ; F, n = 6) in the BAT-negative and BAT-positive patients before (day 0) and after treatment with omalizumab (day 84). Statistical analyses were performed using the Mann-Whitney U test. BAT, basophil activation test.

    Techniques Used: MANN-WHITNEY, Activation Assay

    17) Product Images from "Neutrophils activated by BJcuL, a C-type lectin isolated fromBothrops jararacussu venom, decrease the invasion potential of neuroblastoma SK-N-SH cells in vitro"

    Article Title: Neutrophils activated by BJcuL, a C-type lectin isolated fromBothrops jararacussu venom, decrease the invasion potential of neuroblastoma SK-N-SH cells in vitro

    Journal: The Journal of Venomous Animals and Toxins Including Tropical Diseases

    doi: 10.1590/1678-9199-JVATITD-2019-0073

    Isolation of neutrophils from human peripheral blood. Whole blood underwent a prior centrifugation step to isolate plasma and remove platelets.  (A)  Platelet-poor blood was used for the first fractionation step using Histopaque ®  1077 and 1119, resulting in four distinct layers: plasma, peripheral blood mononuclear cells (PBMC), polymorphonuclear neutrophils (PMN), and red blood cells (RBC). Plasma, PMN and RBC were reunited for a second isolation to allow us to obtain a PMN-enriched fraction. (B)  Flow cytometry analysis of various fractions. Dot plot showing the mixture of PMN, RBC and PBMC in layer 3, and the high concentration of PMN (layer 5).
    Figure Legend Snippet: Isolation of neutrophils from human peripheral blood. Whole blood underwent a prior centrifugation step to isolate plasma and remove platelets. (A) Platelet-poor blood was used for the first fractionation step using Histopaque ® 1077 and 1119, resulting in four distinct layers: plasma, peripheral blood mononuclear cells (PBMC), polymorphonuclear neutrophils (PMN), and red blood cells (RBC). Plasma, PMN and RBC were reunited for a second isolation to allow us to obtain a PMN-enriched fraction. (B) Flow cytometry analysis of various fractions. Dot plot showing the mixture of PMN, RBC and PBMC in layer 3, and the high concentration of PMN (layer 5).

    Techniques Used: Isolation, Centrifugation, Fractionation, Flow Cytometry, Concentration Assay

    18) Product Images from "The CCA-end of P-tRNA Contacts Both the Human RPL36AL and the A-site Bound Translation Termination Factor eRF1 at the Peptidyl Transferase Center of the Human 80S Ribosome"

    Article Title: The CCA-end of P-tRNA Contacts Both the Human RPL36AL and the A-site Bound Translation Termination Factor eRF1 at the Peptidyl Transferase Center of the Human 80S Ribosome

    Journal: The Open Biochemistry Journal

    doi: 10.2174/1874091X01408010052

    Model of the L36AL protein and of its interaction with tRNA. ( A ), ribbon representation of the 3-D structure of human RPL36AL (fragment 1-94) modeled by homology with the crystallographic structure of the archaeal counterpart RPL44E of the 60S ribosomal subunit from Haloarcula marismortui. The post-translational modifications (including the methylated Q51) [ 7 ], and the consensus pattern 61Kx(TorV)KKxxL(KorR)xxC72 (numbering of human RPL36AL) of the L44e family of r-proteins are colored pink and cyan, respectively. The 49GG50 dipeptide of the GGQ motif is highlighted in green. A zinc ion represented by a cadmium colored yellow is also shown. Fragment 86-94 corresponding to the nucleotide binding motif 2 (NBD2) common to all eukaryotic RPL7 [ 29 ] and RPL36A/RPL44 is shown in wheat. ( B ), overlaid structures of tRNAPhe (PDB ID 1JGQ) colored grey and of human RPL36AL (blue). The GGQ motif is shown in red, the 3’ terminal CCA trinucleotide of tRNA in pink, and the side chain of Lys-53 in green. ( C ), NBD2 [ 29 ] located in the C-terminal region of the eukaryotic RPL7 family (ortholog of the bacterial L30 protein) is conserved in the C-terminal region of the eukaryotic RPL36A/RPL44 family. The organisms are: Arabidopsis thaliana (arb. thal.), Drosophila melanogaster (droso.), Saccharomyces cerevisiae (s. cer.), Solanum tuberosum (sol. tub.). The bottom line labels residues as either strictly conserved (*), highly conserved (:) or weakly conserved (.). The alignment was generated with the program ClustalX.
    Figure Legend Snippet: Model of the L36AL protein and of its interaction with tRNA. ( A ), ribbon representation of the 3-D structure of human RPL36AL (fragment 1-94) modeled by homology with the crystallographic structure of the archaeal counterpart RPL44E of the 60S ribosomal subunit from Haloarcula marismortui. The post-translational modifications (including the methylated Q51) [ 7 ], and the consensus pattern 61Kx(TorV)KKxxL(KorR)xxC72 (numbering of human RPL36AL) of the L44e family of r-proteins are colored pink and cyan, respectively. The 49GG50 dipeptide of the GGQ motif is highlighted in green. A zinc ion represented by a cadmium colored yellow is also shown. Fragment 86-94 corresponding to the nucleotide binding motif 2 (NBD2) common to all eukaryotic RPL7 [ 29 ] and RPL36A/RPL44 is shown in wheat. ( B ), overlaid structures of tRNAPhe (PDB ID 1JGQ) colored grey and of human RPL36AL (blue). The GGQ motif is shown in red, the 3’ terminal CCA trinucleotide of tRNA in pink, and the side chain of Lys-53 in green. ( C ), NBD2 [ 29 ] located in the C-terminal region of the eukaryotic RPL7 family (ortholog of the bacterial L30 protein) is conserved in the C-terminal region of the eukaryotic RPL36A/RPL44 family. The organisms are: Arabidopsis thaliana (arb. thal.), Drosophila melanogaster (droso.), Saccharomyces cerevisiae (s. cer.), Solanum tuberosum (sol. tub.). The bottom line labels residues as either strictly conserved (*), highly conserved (:) or weakly conserved (.). The alignment was generated with the program ClustalX.

    Techniques Used: Methylation, Binding Assay, Generated

    Peptidyl-tRNA hydrolase assay of the recombinant human L36AL protein using N-acetyl[3H]Phe-tRNAPhe as a substrate. The percent of residual N-acetyl[3H]Phe-tRNAPhe precipitable in trichloracetic acid was measured as a function of the concentration of the recombinant human RPL36AL. As a control, the activity of E. coli peptidyl-tRNA hydrolase (Pth) as a function of enzyme concentration is also shown.
    Figure Legend Snippet: Peptidyl-tRNA hydrolase assay of the recombinant human L36AL protein using N-acetyl[3H]Phe-tRNAPhe as a substrate. The percent of residual N-acetyl[3H]Phe-tRNAPhe precipitable in trichloracetic acid was measured as a function of the concentration of the recombinant human RPL36AL. As a control, the activity of E. coli peptidyl-tRNA hydrolase (Pth) as a function of enzyme concentration is also shown.

    Techniques Used: Recombinant, Concentration Assay, Activity Assay

    19) Product Images from "PRBC-derived plasma induces non-muscle myosin type IIA-mediated neutrophil migration and morphologic change"

    Article Title: PRBC-derived plasma induces non-muscle myosin type IIA-mediated neutrophil migration and morphologic change

    Journal: Immunopharmacology and Immunotoxicology

    doi: 10.3109/08923973.2012.677046

    Fresh and aged plasmas trigger oxidative burst and protein phosphorylation in neutrophils. (A) Representative results of ROS generation in untreated normal human neutrophils ( Control ), and neutrophils treated for 1 h with fresh (1 day preparation) non-leukocyte-reduced plasma (NLR-1D), aged (42 day preparation) non-leukocyte-reduced plasma (NLR-42D), aged leukocyte-reduced plasma (LR-42D), and NLR-42D plus NADPH oxidase inhibitor DPI (NLR-42D + DPI). The results are expressed as means ± SD from 3 experiments. fMLP: formyl-Met-Leu-Phe (as a positive control). (* p
    Figure Legend Snippet: Fresh and aged plasmas trigger oxidative burst and protein phosphorylation in neutrophils. (A) Representative results of ROS generation in untreated normal human neutrophils ( Control ), and neutrophils treated for 1 h with fresh (1 day preparation) non-leukocyte-reduced plasma (NLR-1D), aged (42 day preparation) non-leukocyte-reduced plasma (NLR-42D), aged leukocyte-reduced plasma (LR-42D), and NLR-42D plus NADPH oxidase inhibitor DPI (NLR-42D + DPI). The results are expressed as means ± SD from 3 experiments. fMLP: formyl-Met-Leu-Phe (as a positive control). (* p

    Techniques Used: Positive Control

    20) Product Images from "Cytoplasmic Trafficking of Minute Virus of Mice: Low-pH Requirement, Routing to Late Endosomes, and Proteasome Interaction"

    Article Title: Cytoplasmic Trafficking of Minute Virus of Mice: Low-pH Requirement, Routing to Late Endosomes, and Proteasome Interaction

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.24.12634-12645.2002

    Effect of TPCK, TLCK, aclarubicin, and PR39 on MVMp infection. A9 cells were infected for 1 h at 37°C with MVMp at an MOI of 10 in the presence of increasing doses of TPCK, TLCK, aclarubicin, or PR39. Subsequently, cells were washed and incubated at 37°C for an additional 1 h in the presence of the irreversible inhibitors TPCK and TLCK or constantly in the case of the reversible drugs aclarubicin and PR39. Cells were then washed, and the amount of viral DNA was quantified 18 h postinfection. Values represent the mean of three independent experiments. nt, nontreated; ni, noninfected.
    Figure Legend Snippet: Effect of TPCK, TLCK, aclarubicin, and PR39 on MVMp infection. A9 cells were infected for 1 h at 37°C with MVMp at an MOI of 10 in the presence of increasing doses of TPCK, TLCK, aclarubicin, or PR39. Subsequently, cells were washed and incubated at 37°C for an additional 1 h in the presence of the irreversible inhibitors TPCK and TLCK or constantly in the case of the reversible drugs aclarubicin and PR39. Cells were then washed, and the amount of viral DNA was quantified 18 h postinfection. Values represent the mean of three independent experiments. nt, nontreated; ni, noninfected.

    Techniques Used: Infection, Incubation

    21) Product Images from "Genotoxic mixtures and dissimilar action: concepts for prediction and assessment"

    Article Title: Genotoxic mixtures and dissimilar action: concepts for prediction and assessment

    Journal: Archives of Toxicology

    doi: 10.1007/s00204-013-1170-x

    Induction of MN by aneugens and clastogens in the CBMN assay using CHO-K1 cells. MN induction is presented as percentage of MN positive binucleated cells. The graphs show the data for at least three independent experiments ( red dots , exception: benzo[α]pyrene was tested only once); solvent controls are shown on the left ( green dots as indicated). The regression curves ( thick black lines ) are shown with their 95 % confidence belts ( dashed lines ). Estimated threshold concentrations are indicated by the vertical dashed lines . Mean baseline levels of MN within the cells are depicted as horizontal lines . The grey areas show the cytotoxic concentrations determined in the MTT assay (MTT–EC 40 ) (colour figure online)
    Figure Legend Snippet: Induction of MN by aneugens and clastogens in the CBMN assay using CHO-K1 cells. MN induction is presented as percentage of MN positive binucleated cells. The graphs show the data for at least three independent experiments ( red dots , exception: benzo[α]pyrene was tested only once); solvent controls are shown on the left ( green dots as indicated). The regression curves ( thick black lines ) are shown with their 95 % confidence belts ( dashed lines ). Estimated threshold concentrations are indicated by the vertical dashed lines . Mean baseline levels of MN within the cells are depicted as horizontal lines . The grey areas show the cytotoxic concentrations determined in the MTT assay (MTT–EC 40 ) (colour figure online)

    Techniques Used: MTT Assay

    22) Product Images from "Finasteride inhibits melanogenesis through regulation of the adenylate cyclase in melanocytes and melanoma cells"

    Article Title: Finasteride inhibits melanogenesis through regulation of the adenylate cyclase in melanocytes and melanoma cells

    Journal: Archives of Pharmacal Research

    doi: 10.1007/s12272-018-1002-x

    Inhibitory effects of finasteride on DOPA and tyrosinase activity. a Melan-a cells were incubated in the absence or b presence of 10 µM finasteride. c B16F10 cells were incubated without treatment, d with 100 nM α-MSH, or with e 100 µM finasteride and 100 nM α-MSH. f Untreated NHEM cells and g cells treated with 10 µM finasteride. h Inhibitory effects of finasteride on mushroom tyrosinase activity. i Inhibitory effects of finasteride on tyrosinase activity in melan-a cells. Cells were treated with finasteride (0.1, 1, or 10 µM) and the amount of DOPA-chrome was measured. Kojic acid was used as a positive control. All data are expressed as the mean ± SEM, and were analyzed by one-way ANOVA, followed by the Student’s t test. *p
    Figure Legend Snippet: Inhibitory effects of finasteride on DOPA and tyrosinase activity. a Melan-a cells were incubated in the absence or b presence of 10 µM finasteride. c B16F10 cells were incubated without treatment, d with 100 nM α-MSH, or with e 100 µM finasteride and 100 nM α-MSH. f Untreated NHEM cells and g cells treated with 10 µM finasteride. h Inhibitory effects of finasteride on mushroom tyrosinase activity. i Inhibitory effects of finasteride on tyrosinase activity in melan-a cells. Cells were treated with finasteride (0.1, 1, or 10 µM) and the amount of DOPA-chrome was measured. Kojic acid was used as a positive control. All data are expressed as the mean ± SEM, and were analyzed by one-way ANOVA, followed by the Student’s t test. *p

    Techniques Used: Activity Assay, Incubation, Positive Control

    Inhibitory effects of finasteride on melanogenic enzyme expressions. a Melan-a cells were treated with finasteride (0.1, 1, or 10 µM) for 72 h. b B16F10 cells were co-treated with 100 nM α-MSH and indicated concentration of finasteride. Equal amounts of protein (40 µg/lane) were separated using a 12% SDS–PAGE gel, and were detected using specific antibodies. Equal protein loading was evaluated using antibodies against α-tubulin
    Figure Legend Snippet: Inhibitory effects of finasteride on melanogenic enzyme expressions. a Melan-a cells were treated with finasteride (0.1, 1, or 10 µM) for 72 h. b B16F10 cells were co-treated with 100 nM α-MSH and indicated concentration of finasteride. Equal amounts of protein (40 µg/lane) were separated using a 12% SDS–PAGE gel, and were detected using specific antibodies. Equal protein loading was evaluated using antibodies against α-tubulin

    Techniques Used: Concentration Assay, SDS Page

    Inhibitory effects of finasteride on melanin contents and cell viability in Melan-a and B16F10 cells. Cells were treated with the indicated concentration of finasteride for 72 h. a Melanin content and b cell growth rate were measured in melan-a cells. c Amount of melanin and d cell growth rate in B16F10 cells with µ nM of α-MSH. White bar represent untreated cells and black bars represent α-MSH-treated cells. All data are expressed as mean ± SEM, and were analyzed by one-way ANOVA, followed by the Student’s t test. *p
    Figure Legend Snippet: Inhibitory effects of finasteride on melanin contents and cell viability in Melan-a and B16F10 cells. Cells were treated with the indicated concentration of finasteride for 72 h. a Melanin content and b cell growth rate were measured in melan-a cells. c Amount of melanin and d cell growth rate in B16F10 cells with µ nM of α-MSH. White bar represent untreated cells and black bars represent α-MSH-treated cells. All data are expressed as mean ± SEM, and were analyzed by one-way ANOVA, followed by the Student’s t test. *p

    Techniques Used: Concentration Assay

    23) Product Images from "Artichoke Polyphenols Produce Skin Anti-Age Effects by Improving Endothelial Cell Integrity and Functionality"

    Article Title: Artichoke Polyphenols Produce Skin Anti-Age Effects by Improving Endothelial Cell Integrity and Functionality

    Journal: Molecules

    doi: 10.3390/molecules23112729

    Protection against inflammation. The levels of NO production ( a ) and iNOS gene expression ( b ) were measured in RAW 264.7 macrophages treated with AEE before addition of bacterial liposaccharide LPS. TPCK was used as positive control. All determinations were conducted in triplicate and results are expressed as mean ± SD. * p
    Figure Legend Snippet: Protection against inflammation. The levels of NO production ( a ) and iNOS gene expression ( b ) were measured in RAW 264.7 macrophages treated with AEE before addition of bacterial liposaccharide LPS. TPCK was used as positive control. All determinations were conducted in triplicate and results are expressed as mean ± SD. * p

    Techniques Used: Expressing, Positive Control

    24) Product Images from "Protection of Bcl-2 by salubrinal"

    Article Title: Protection of Bcl-2 by salubrinal

    Journal:

    doi: 10.1016/j.bbrc.2006.06.056

    Western blots showing: (A) levels of eIF2α (phosphorylated and non-phosphorylated) 60 min after exposure of L1210 cells to 50 μM salubrinal or 60 min after treatment with an LD 90 concentration of HA14-1. Salubrinal was present where specified.
    Figure Legend Snippet: Western blots showing: (A) levels of eIF2α (phosphorylated and non-phosphorylated) 60 min after exposure of L1210 cells to 50 μM salubrinal or 60 min after treatment with an LD 90 concentration of HA14-1. Salubrinal was present where specified.

    Techniques Used: Western Blot, Concentration Assay

    25) Product Images from "Purification and Characterization of an Extracellular, Thermo-Alkali-Stable, Metal Tolerant Laccase from Bacillus tequilensis SN4"

    Article Title: Purification and Characterization of an Extracellular, Thermo-Alkali-Stable, Metal Tolerant Laccase from Bacillus tequilensis SN4

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0096951

    SDS-PAGE analysis of SN4LAC from B. tequilensis SN4. (Protein samples were denatured by heating for 5 min in the presence of SDS and β-mercaptoethanol): Lane 1: Protein markers, Lane 2: Acetone precipitated proteins, Lane 3: Sephadex-150 Column purified enzyme, lane 4: DEAE-Cellulose anion exchange Column purified enzyme, Lane 5: Activity staining; purified laccase stained with guaiacol (samples were heated for 5 min in the presence of SDS without β-mercaptoethanol).
    Figure Legend Snippet: SDS-PAGE analysis of SN4LAC from B. tequilensis SN4. (Protein samples were denatured by heating for 5 min in the presence of SDS and β-mercaptoethanol): Lane 1: Protein markers, Lane 2: Acetone precipitated proteins, Lane 3: Sephadex-150 Column purified enzyme, lane 4: DEAE-Cellulose anion exchange Column purified enzyme, Lane 5: Activity staining; purified laccase stained with guaiacol (samples were heated for 5 min in the presence of SDS without β-mercaptoethanol).

    Techniques Used: SDS Page, Purification, Activity Assay, Staining

    26) Product Images from "Time-Resolved Infrared Spectroscopy of RNA Folding"

    Article Title: Time-Resolved Infrared Spectroscopy of RNA Folding

    Journal: Biophysical Journal

    doi: 10.1529/biophysj.105.061531

    FTIR ( a ) and thermal difference spectra ( b ) of tRNA phe . ( a ) Folded ( dashed line ) and unfolded ( solid line ); the symbols mark the positions of the main vibrational modes and their assignments. ( b ) The low-temperature spectrum is subtracted from all other spectra. The arrows indicate the direction of spectral changes with increasing temperature. Temperatures ranged from 5.5 to 86.6°C in ∼5°C increments. Absorption changes at 1620 cm −1 correspond to A and U interactions and the changes at 1661 cm −1 correspond to G and C interactions. All of the spectral features in this wavelength range are due to C=N, C=C, and C=O stretching vibrations of the nucleic acid bases.
    Figure Legend Snippet: FTIR ( a ) and thermal difference spectra ( b ) of tRNA phe . ( a ) Folded ( dashed line ) and unfolded ( solid line ); the symbols mark the positions of the main vibrational modes and their assignments. ( b ) The low-temperature spectrum is subtracted from all other spectra. The arrows indicate the direction of spectral changes with increasing temperature. Temperatures ranged from 5.5 to 86.6°C in ∼5°C increments. Absorption changes at 1620 cm −1 correspond to A and U interactions and the changes at 1661 cm −1 correspond to G and C interactions. All of the spectral features in this wavelength range are due to C=N, C=C, and C=O stretching vibrations of the nucleic acid bases.

    Techniques Used:

    27) Product Images from "Gene silencing of ?-galactosamide ?-2,6-sialyltransferase 1 inhibits human influenza virus infection of airway epithelial cells"

    Article Title: Gene silencing of ?-galactosamide ?-2,6-sialyltransferase 1 inhibits human influenza virus infection of airway epithelial cells

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-14-78

    ST6GAL1-specific siRNAs inhibited ST6Gal 1 expression and SAα2,6Gal synthesis. Respiratory epithelial cells were transfected with control or ST6GAL1 siRNAs (A) At 48 h post-transfection, ST6GAL1 mRNA expression levels were quantified. (B) Candidate siRNAs reduce ST6GAL1 expression in a dose-dependent manner. (C) FACS analysis showed a decrease in SAα2,6Gal expression on ST6GAL1 siRNA-treated cell surfaces. (D) Inhibition of ST6Gal Ι expression due to ST6GAL1 siRNAs. (E) We used ST6GAL1 siRNAs at various concentrations and they were not cytotoxic, except at 50 nM. a P
    Figure Legend Snippet: ST6GAL1-specific siRNAs inhibited ST6Gal 1 expression and SAα2,6Gal synthesis. Respiratory epithelial cells were transfected with control or ST6GAL1 siRNAs (A) At 48 h post-transfection, ST6GAL1 mRNA expression levels were quantified. (B) Candidate siRNAs reduce ST6GAL1 expression in a dose-dependent manner. (C) FACS analysis showed a decrease in SAα2,6Gal expression on ST6GAL1 siRNA-treated cell surfaces. (D) Inhibition of ST6Gal Ι expression due to ST6GAL1 siRNAs. (E) We used ST6GAL1 siRNAs at various concentrations and they were not cytotoxic, except at 50 nM. a P

    Techniques Used: Expressing, Transfection, FACS, Inhibition

    ST6GAL1 siRNA-transduced respiratory cells resisted human influenza virus challenge and did not induce an interferon response. Transduced A549 cells were challenged with H3N2, pdmH1N1, or H9N2 viruses. (A) A reduction in viral yield was seen in ST6GAL1 siRNA-transduced cells infected with and pdmH1N1 (B) H3N2 influenza viruses. a P
    Figure Legend Snippet: ST6GAL1 siRNA-transduced respiratory cells resisted human influenza virus challenge and did not induce an interferon response. Transduced A549 cells were challenged with H3N2, pdmH1N1, or H9N2 viruses. (A) A reduction in viral yield was seen in ST6GAL1 siRNA-transduced cells infected with and pdmH1N1 (B) H3N2 influenza viruses. a P

    Techniques Used: Infection

    28) Product Images from "STRUCTURE OF THE EGF RECEPTOR TRANSACTIVATION CIRCUIT INTEGRATES MULTIPLE SIGNALS WITH CELL CONTEXT"

    Article Title: STRUCTURE OF THE EGF RECEPTOR TRANSACTIVATION CIRCUIT INTEGRATES MULTIPLE SIGNALS WITH CELL CONTEXT

    Journal: Molecular Biosystems

    doi: 10.1039/c003921g

    EGFR autocrine ligands stimulate their own release by activating ERK ( A ) Accumulation of EGF in medium is inhibited by blocking the EGFR or inhibiting cell surface proteolysis. After 16 hours in serum-free media, cells expressing the chimeric ligand TCT were switched to fresh serum-free media alone or containing 10 μg/ml 225 mAb or 225 mAb in addition to 10 μM batimastat. Media from triplicate wells was collected at 2-hour intervals, assayed for EGF with an ELISA. Error bars represent standard deviation. ( B ) Effects of inhibitors on ligand release. After 16 hours in serum-free media, TCT cells were pre-incubated with the indicated inhibitors for 30 minutes. Cells were then switched to either fresh serum-free media alone (control) or containing 1 μM Gefitinib, 10 μM SB203580 or 25 μM PD98059. Other conditions same as A. ( C ) Cells expressing TCT-NA were switched to fresh serum-free medium at t=0, and the amount of immunoreactive EGF in the medium was determined by ELISA (open circles). At 1hr, batimastat was added to a final concentration of 10μM to a parallel set of cells (closed circles). ( D ) Blocking the EGFR has no significant effect on release of TCT-NA. Cells expressing TCT-NA were changed to serum-free medium either in the absence (open bars) or presence (closed bars) of 10μg/ml of 225 mAb. After 8 hr, the medium was collected and the concentration of immunoreactive EGF was measured by ELISA. Error bars are SD from triplicate wells. ( E ) TCT-NA fails to activate the EGFR. Serum-free conditioned medium was collected from an overnight incubation of WT HMEC (control medium) or from cells expressing either TCT or TCT-NA. Immunoreactive levels of EGF were determined by ELISA and the final concentrations of both ligands were adjusted to 5ng/ml. Parental B82 mouse fibroblasts lacking the EGFR (R-) or transfected with the human EGFR (R+) were incubated with the indicated medium for 10 min. The levels of phospho-EGFR were measured by ELISA. Error bars are SD from triplicate wells. ( F ) Shedding of the non-bindable EGFR ligand TCT-NA is stimulated by TGFα and inhibited by ERK inhibitors. After 16 hours in serum-free media, cells were switched to fresh serum-free media alone or medium containing 20 ng/ml TGFα, both in the presence or absence of 25 μM PD98059. Other conditions same as A.
    Figure Legend Snippet: EGFR autocrine ligands stimulate their own release by activating ERK ( A ) Accumulation of EGF in medium is inhibited by blocking the EGFR or inhibiting cell surface proteolysis. After 16 hours in serum-free media, cells expressing the chimeric ligand TCT were switched to fresh serum-free media alone or containing 10 μg/ml 225 mAb or 225 mAb in addition to 10 μM batimastat. Media from triplicate wells was collected at 2-hour intervals, assayed for EGF with an ELISA. Error bars represent standard deviation. ( B ) Effects of inhibitors on ligand release. After 16 hours in serum-free media, TCT cells were pre-incubated with the indicated inhibitors for 30 minutes. Cells were then switched to either fresh serum-free media alone (control) or containing 1 μM Gefitinib, 10 μM SB203580 or 25 μM PD98059. Other conditions same as A. ( C ) Cells expressing TCT-NA were switched to fresh serum-free medium at t=0, and the amount of immunoreactive EGF in the medium was determined by ELISA (open circles). At 1hr, batimastat was added to a final concentration of 10μM to a parallel set of cells (closed circles). ( D ) Blocking the EGFR has no significant effect on release of TCT-NA. Cells expressing TCT-NA were changed to serum-free medium either in the absence (open bars) or presence (closed bars) of 10μg/ml of 225 mAb. After 8 hr, the medium was collected and the concentration of immunoreactive EGF was measured by ELISA. Error bars are SD from triplicate wells. ( E ) TCT-NA fails to activate the EGFR. Serum-free conditioned medium was collected from an overnight incubation of WT HMEC (control medium) or from cells expressing either TCT or TCT-NA. Immunoreactive levels of EGF were determined by ELISA and the final concentrations of both ligands were adjusted to 5ng/ml. Parental B82 mouse fibroblasts lacking the EGFR (R-) or transfected with the human EGFR (R+) were incubated with the indicated medium for 10 min. The levels of phospho-EGFR were measured by ELISA. Error bars are SD from triplicate wells. ( F ) Shedding of the non-bindable EGFR ligand TCT-NA is stimulated by TGFα and inhibited by ERK inhibitors. After 16 hours in serum-free media, cells were switched to fresh serum-free media alone or medium containing 20 ng/ml TGFα, both in the presence or absence of 25 μM PD98059. Other conditions same as A.

    Techniques Used: Blocking Assay, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, Incubation, Concentration Assay, Transfection

    29) Product Images from "Mechanisms Mediating Reduced Responsiveness of Neonatal Neutrophils to Lipoxin A4"

    Article Title: Mechanisms Mediating Reduced Responsiveness of Neonatal Neutrophils to Lipoxin A4

    Journal: Pediatric research

    doi: 10.1203/PDR.0b013e318180e4af

    Effects of lipoxin A4 concentration on induction of PPAR-γ
    Figure Legend Snippet: Effects of lipoxin A4 concentration on induction of PPAR-γ

    Techniques Used: Concentration Assay

    Effects of lipoxin A4 on neutrophil respiratory burst activity
    Figure Legend Snippet: Effects of lipoxin A4 on neutrophil respiratory burst activity

    Techniques Used: Activity Assay

    Effects of lipoxin A4 on expression of PPAR-γ and NGAL
    Figure Legend Snippet: Effects of lipoxin A4 on expression of PPAR-γ and NGAL

    Techniques Used: Expressing

    Effects of lipoxin A4 on neutrophil chemotaxis and apoptosis
    Figure Legend Snippet: Effects of lipoxin A4 on neutrophil chemotaxis and apoptosis

    Techniques Used: Chemotaxis Assay

    30) Product Images from "Response of Midgut Trypsin- and Chymotrypsin-Like Proteases of Helicoverpa armigera Larvae Upon Feeding With Peanut BBI: Biochemical and Biophysical Characterization of PnBBI"

    Article Title: Response of Midgut Trypsin- and Chymotrypsin-Like Proteases of Helicoverpa armigera Larvae Upon Feeding With Peanut BBI: Biochemical and Biophysical Characterization of PnBBI

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2020.00266

    Purification profile of PnBBI and its in-gel activity. Elution profile of (A) trypsin-Sepharose 4B column loaded with 20-60% (NH 4 ) 2 SO 4 active fraction; (B) Sephadex G-50 fine column loaded with active peak II fraction pool of trypsin affinity column; (C) Tricine SDS-PAGE (15%) showing purification profile and self-association pattern of PnBBI: lane 1, molecular weight marker designated in kDa; lane 2, peanut crude protein extract (20 μg); lane 3, 20-60% (NH 4 ) 2 SO 4 protein fraction (15 μg); lane 4, active fraction pool (Peak II) of trypsin affinity column (10 μg); lanes 5-7 and 8-10 active fraction pool (peak II) of gel filtration column under non-reducing and reducing conditions with increased protein concentration (2.5, 5, and 10 μg), respectively; lane 11, soybean BBI (5 μg) was used as a reference. Gelatin SDS-PAGE (15%): Lane 1, soybean BBI (5 μg); lane 2, PnBBI (5 μg) active against (D) bovine pancreatic trypsin and (E) chymotrypsin, respectively. Asterisks indicate active peak with inhibitory activity against trypsin. M-monomer, D-dimer and T-tetramer. The data shown here is the representative of three biological replicates (data provided as Supplementary Data Sheet 3 ).
    Figure Legend Snippet: Purification profile of PnBBI and its in-gel activity. Elution profile of (A) trypsin-Sepharose 4B column loaded with 20-60% (NH 4 ) 2 SO 4 active fraction; (B) Sephadex G-50 fine column loaded with active peak II fraction pool of trypsin affinity column; (C) Tricine SDS-PAGE (15%) showing purification profile and self-association pattern of PnBBI: lane 1, molecular weight marker designated in kDa; lane 2, peanut crude protein extract (20 μg); lane 3, 20-60% (NH 4 ) 2 SO 4 protein fraction (15 μg); lane 4, active fraction pool (Peak II) of trypsin affinity column (10 μg); lanes 5-7 and 8-10 active fraction pool (peak II) of gel filtration column under non-reducing and reducing conditions with increased protein concentration (2.5, 5, and 10 μg), respectively; lane 11, soybean BBI (5 μg) was used as a reference. Gelatin SDS-PAGE (15%): Lane 1, soybean BBI (5 μg); lane 2, PnBBI (5 μg) active against (D) bovine pancreatic trypsin and (E) chymotrypsin, respectively. Asterisks indicate active peak with inhibitory activity against trypsin. M-monomer, D-dimer and T-tetramer. The data shown here is the representative of three biological replicates (data provided as Supplementary Data Sheet 3 ).

    Techniques Used: Purification, Activity Assay, Affinity Column, SDS Page, Molecular Weight, Marker, Filtration, Protein Concentration

    31) Product Images from "IL-2-induced expression of the amino acid transporters SLC1A5 and CD98 is a prerequisite for NKG2D-mediated activation of human NK cells"

    Article Title: IL-2-induced expression of the amino acid transporters SLC1A5 and CD98 is a prerequisite for NKG2D-mediated activation of human NK cells

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1700497

    Inhibition of mTORC1 activity abrogates NKG2D-induced IFNγ production NK cells enriched from human PBMCs were primed with 200 U/ml IL-2 for 24 h. Cells were treated with medium, 100 nM rapamycin, or 100 nM Torin-1 for 1 h prior to stimulation with plate-bound anti-NKG2D Abs for 5 h. IFNγ production was measured in CD56 bright and CD56 dim NK cells by flow cytometry. ( A ) Results are shown for one donor and are representative of three donors. ( B ) Data show mean ± s.d. from three donors. Statistical analysis was performed by 2-tailed unpaired student's t-test (*p
    Figure Legend Snippet: Inhibition of mTORC1 activity abrogates NKG2D-induced IFNγ production NK cells enriched from human PBMCs were primed with 200 U/ml IL-2 for 24 h. Cells were treated with medium, 100 nM rapamycin, or 100 nM Torin-1 for 1 h prior to stimulation with plate-bound anti-NKG2D Abs for 5 h. IFNγ production was measured in CD56 bright and CD56 dim NK cells by flow cytometry. ( A ) Results are shown for one donor and are representative of three donors. ( B ) Data show mean ± s.d. from three donors. Statistical analysis was performed by 2-tailed unpaired student's t-test (*p

    Techniques Used: Inhibition, Activity Assay, Flow Cytometry, Cytometry

    32) Product Images from "Closing and opening of the RNA polymerase trigger loop"

    Article Title: Closing and opening of the RNA polymerase trigger loop

    Journal: bioRxiv

    doi: 10.1101/817080

    Incorporation of fluorescent probe into RNAP: singly-labelled RNAP derivative a, Products of labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit with Dylight 550 phosphine, as detected by Coomassie staining (left) and fluorescent scanning in donor-emission channel (532 nm excitation and 580 nm emission bandpass filters; right). Fluorescent labelling is observed only for the β’ subunit. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.
    Figure Legend Snippet: Incorporation of fluorescent probe into RNAP: singly-labelled RNAP derivative a, Products of labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit with Dylight 550 phosphine, as detected by Coomassie staining (left) and fluorescent scanning in donor-emission channel (532 nm excitation and 580 nm emission bandpass filters; right). Fluorescent labelling is observed only for the β’ subunit. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.

    Techniques Used: Staining

    Incorporation of fluorescent probes into RNAP: doubly-labelled RNAP derivative a, Products of stochastic labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit and position 267 of β subunit with Dylight 550 phosphine and Dylight 650 phosphine, as detected by Coomassie staining (left), fluorescent scanning in donor-emission channel (D; 532 nm excitation and 580 nm emission bandpass filters; center) and fluorescent scanning in acceptor-emission channel (A; 633 nm excitation and 670 nm emission bandpass filters; right). Fluorescent labelling is observed only for β’ and β subunits. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.
    Figure Legend Snippet: Incorporation of fluorescent probes into RNAP: doubly-labelled RNAP derivative a, Products of stochastic labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit and position 267 of β subunit with Dylight 550 phosphine and Dylight 650 phosphine, as detected by Coomassie staining (left), fluorescent scanning in donor-emission channel (D; 532 nm excitation and 580 nm emission bandpass filters; center) and fluorescent scanning in acceptor-emission channel (A; 633 nm excitation and 670 nm emission bandpass filters; right). Fluorescent labelling is observed only for β’ and β subunits. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.

    Techniques Used: Staining

    33) Product Images from "Closing and opening of the RNA polymerase trigger loop"

    Article Title: Closing and opening of the RNA polymerase trigger loop

    Journal: bioRxiv

    doi: 10.1101/817080

    One TL closing-opening cycle typically occurs in each nucleotide addition in transcription elongation. a,  Use of unnatural-amino-acid mutagenesis (first reaction arrow), Staudinger ligation with Dylight 550-phosphine and Dylight 650-phosphine (second reaction arrow), TEC reconstitution (third reaction arrow), and total-internal-reflection fluorescence microscopy with alternating-laser excitation microscopy (TIRF-ALEX) to measure smFRET between first fluorescent probe at tip of RNAP TL and second fluorescent probe at reference site elsewhere in RNAP (see Methods). Green filled circles, fluorescent probe Dylight 550; orange filled circles, fluorescent probe Dylight 650. Other colors as in   Fig. 1c . b,  smFRET data for TEC engaged in active real-time transcription elongation on templates directing addition of 1, 2, 3, or 4 additions of A. Left subpanels, representative time traces of donor-acceptor FRET, E*. Black dashed line, addition of ATP. Other colors as in left bottom subpanel of   Fig. 2a . Right subpanels, histograms showing numbers of detected TL closing/opening cycles. Note one-for-one correlation between the modal number of TL closing/opening events detected for a template (dark gray bar for each template) and the number of nucleotide additions directed by the template.
    Figure Legend Snippet: One TL closing-opening cycle typically occurs in each nucleotide addition in transcription elongation. a, Use of unnatural-amino-acid mutagenesis (first reaction arrow), Staudinger ligation with Dylight 550-phosphine and Dylight 650-phosphine (second reaction arrow), TEC reconstitution (third reaction arrow), and total-internal-reflection fluorescence microscopy with alternating-laser excitation microscopy (TIRF-ALEX) to measure smFRET between first fluorescent probe at tip of RNAP TL and second fluorescent probe at reference site elsewhere in RNAP (see Methods). Green filled circles, fluorescent probe Dylight 550; orange filled circles, fluorescent probe Dylight 650. Other colors as in Fig. 1c . b, smFRET data for TEC engaged in active real-time transcription elongation on templates directing addition of 1, 2, 3, or 4 additions of A. Left subpanels, representative time traces of donor-acceptor FRET, E*. Black dashed line, addition of ATP. Other colors as in left bottom subpanel of Fig. 2a . Right subpanels, histograms showing numbers of detected TL closing/opening cycles. Note one-for-one correlation between the modal number of TL closing/opening events detected for a template (dark gray bar for each template) and the number of nucleotide additions directed by the template.

    Techniques Used: Mutagenesis, Ligation, Fluorescence, Microscopy

    Incorporation of fluorescent probe into RNAP: singly-labelled RNAP derivative a, Products of labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit with Dylight 550 phosphine, as detected by Coomassie staining (left) and fluorescent scanning in donor-emission channel (532 nm excitation and 580 nm emission bandpass filters; right). Fluorescent labelling is observed only for the β’ subunit. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.
    Figure Legend Snippet: Incorporation of fluorescent probe into RNAP: singly-labelled RNAP derivative a, Products of labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit with Dylight 550 phosphine, as detected by Coomassie staining (left) and fluorescent scanning in donor-emission channel (532 nm excitation and 580 nm emission bandpass filters; right). Fluorescent labelling is observed only for the β’ subunit. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.

    Techniques Used: Staining

    Incorporation of fluorescent probes into RNAP: doubly-labelled RNAP derivative a, Products of stochastic labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit and position 267 of β subunit with Dylight 550 phosphine and Dylight 650 phosphine, as detected by Coomassie staining (left), fluorescent scanning in donor-emission channel (D; 532 nm excitation and 580 nm emission bandpass filters; center) and fluorescent scanning in acceptor-emission channel (A; 633 nm excitation and 670 nm emission bandpass filters; right). Fluorescent labelling is observed only for β’ and β subunits. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.
    Figure Legend Snippet: Incorporation of fluorescent probes into RNAP: doubly-labelled RNAP derivative a, Products of stochastic labelling reaction of RNAP derivative containing 4-azidophenylalanine (AzF) at position 942 of β’ subunit and position 267 of β subunit with Dylight 550 phosphine and Dylight 650 phosphine, as detected by Coomassie staining (left), fluorescent scanning in donor-emission channel (D; 532 nm excitation and 580 nm emission bandpass filters; center) and fluorescent scanning in acceptor-emission channel (A; 633 nm excitation and 670 nm emission bandpass filters; right). Fluorescent labelling is observed only for β’ and β subunits. b, Labelling efficiencies and labelling specificities (see Materials and Methods). c, Transcriptional activities of fluorescent-probe-labelled RNAP.

    Techniques Used: Staining

    Use of smFRET to detect and characterize TL closing and opening in solution a, Open-TL (left subpanel) and closed-TL (right subpanel) conformational states as observed in crystal structures of T. thermophilus RNAP ( 9,29 ; PDB 1ZYR and PDB 2O5J). Gray and red ribbon, RNAP trigger helices and TL in open-TL state; gray and green ribbon, RNAP trigger helices and TL in closed-TL state, gray and black sticks, DNA non-template and template strands; blue sticks, RNA 3’ nucleotide; cyan stick, incoming NTP; purple sphere in left panel, catalytic Mg 2+ ions Mg 2+ (I) (left and right subpanels) and Mg 2+ (II) (right subpanel). b , Measurement of smFRET between first fluorescent probe incorporated at tip of RNAP TL (red sphere for open-TL state; green sphere for closed-TL) and second fluorescent probe incorporated into downstream DNA (pink sphere). Estimated probe-probe distances are ~34 Å for open-TL state and ~39 Å for closed-TL state. c, Use of unnatural--amino-acid mutagenesis (first reaction arrow), Staudinger ligation (second reaction arrow), and TEC reconstitution (third reaction arrow) to prepare sample for measurement of smFRET between first fluorescent probe incorporated at tip of RNAP TL and second fluorescent probe incorporated into downstream DNA (see Methods). Black open ovals, open rectangles, two-segment arrows, arrowhead labelled “amber,” and arrowhead labelled “His 6 ” denote plasmids, genes, promoters, amber codon in TL coding sequence in gene for RNAP β’ subunit, and hexahistidine coding sequence at 3’ end of gene for RNAP β’ subunit. Gray ribbon structure, black open circle, green filled circle, and black bar denote RNAP, unnatural amino acid 4-azidophenylalanine in RNAP TL, fluorescent probe Dylight 550 in RNAP TL, and hexahistidine tag at C-terminus of RNAP β’ subunit. Gray lines, black lines, blue lines, and orange filled circle denote nucleic-acid scaffold comprising nontemplate-strand DNA, template-strand DNA, RNA, and fluorescent probe Alexa647. d, smFRET data for TEC in post-translocated state in absence of NTP (top) and in presence of saturating concentration of complementary NTP (1 mM ATP; bottom). Left subpanels, representative time traces of donor-acceptor FRET efficiency, E*, showing open-TL (top, red) and closed-TL (bottom, green) states. Right subpanel, histograms and Gaussian fits of E*, showing mean E* values for open-TL (red line) and closed-TL (green line) states; n, number of frames.
    Figure Legend Snippet: Use of smFRET to detect and characterize TL closing and opening in solution a, Open-TL (left subpanel) and closed-TL (right subpanel) conformational states as observed in crystal structures of T. thermophilus RNAP ( 9,29 ; PDB 1ZYR and PDB 2O5J). Gray and red ribbon, RNAP trigger helices and TL in open-TL state; gray and green ribbon, RNAP trigger helices and TL in closed-TL state, gray and black sticks, DNA non-template and template strands; blue sticks, RNA 3’ nucleotide; cyan stick, incoming NTP; purple sphere in left panel, catalytic Mg 2+ ions Mg 2+ (I) (left and right subpanels) and Mg 2+ (II) (right subpanel). b , Measurement of smFRET between first fluorescent probe incorporated at tip of RNAP TL (red sphere for open-TL state; green sphere for closed-TL) and second fluorescent probe incorporated into downstream DNA (pink sphere). Estimated probe-probe distances are ~34 Å for open-TL state and ~39 Å for closed-TL state. c, Use of unnatural--amino-acid mutagenesis (first reaction arrow), Staudinger ligation (second reaction arrow), and TEC reconstitution (third reaction arrow) to prepare sample for measurement of smFRET between first fluorescent probe incorporated at tip of RNAP TL and second fluorescent probe incorporated into downstream DNA (see Methods). Black open ovals, open rectangles, two-segment arrows, arrowhead labelled “amber,” and arrowhead labelled “His 6 ” denote plasmids, genes, promoters, amber codon in TL coding sequence in gene for RNAP β’ subunit, and hexahistidine coding sequence at 3’ end of gene for RNAP β’ subunit. Gray ribbon structure, black open circle, green filled circle, and black bar denote RNAP, unnatural amino acid 4-azidophenylalanine in RNAP TL, fluorescent probe Dylight 550 in RNAP TL, and hexahistidine tag at C-terminus of RNAP β’ subunit. Gray lines, black lines, blue lines, and orange filled circle denote nucleic-acid scaffold comprising nontemplate-strand DNA, template-strand DNA, RNA, and fluorescent probe Alexa647. d, smFRET data for TEC in post-translocated state in absence of NTP (top) and in presence of saturating concentration of complementary NTP (1 mM ATP; bottom). Left subpanels, representative time traces of donor-acceptor FRET efficiency, E*, showing open-TL (top, red) and closed-TL (bottom, green) states. Right subpanel, histograms and Gaussian fits of E*, showing mean E* values for open-TL (red line) and closed-TL (green line) states; n, number of frames.

    Techniques Used: Mutagenesis, Ligation, Sequencing, Concentration Assay

    34) Product Images from "Protective effects of flavonol isoquercitrin, against 6-hydroxy dopamine (6-OHDA) - induced toxicity in PC12 cells"

    Article Title: Protective effects of flavonol isoquercitrin, against 6-hydroxy dopamine (6-OHDA) - induced toxicity in PC12 cells

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-7-49

    The dose–response of different concentrations of 6-hydroxydopamine (6-OHDA) (0–200 μM) used to induce neurodegeneration in 1 × 10 5 cells/well. Pheochromocytoma (PC 12) cells for the duration of 24 h. The viability of PC12 cells was determined using the MTT reduction assay. Values are the percentages of viable cells, with the viability of untreated control cells taken as 100%. Data are mean and S.E. values from three independent experiments (n = 4). ** p
    Figure Legend Snippet: The dose–response of different concentrations of 6-hydroxydopamine (6-OHDA) (0–200 μM) used to induce neurodegeneration in 1 × 10 5 cells/well. Pheochromocytoma (PC 12) cells for the duration of 24 h. The viability of PC12 cells was determined using the MTT reduction assay. Values are the percentages of viable cells, with the viability of untreated control cells taken as 100%. Data are mean and S.E. values from three independent experiments (n = 4). ** p

    Techniques Used: MTT Assay

    35) Product Images from "Effects of Five Ayurvedic Herbs on Locomotor Behaviour in a Drosophila melanogaster Parkinson’s Disease Model"

    Article Title: Effects of Five Ayurvedic Herbs on Locomotor Behaviour in a Drosophila melanogaster Parkinson’s Disease Model

    Journal: Phytotherapy research : PTR

    doi: 10.1002/ptr.5199

    Effects of different herbs or herbal preparations on the percentage of flies that escaped in the climbing ability test for flies with a loss of function PINK1 gene (PINK1) and wild-type (WT) flies. Vitamin C was added to levodopa to prevent oxidation.
    Figure Legend Snippet: Effects of different herbs or herbal preparations on the percentage of flies that escaped in the climbing ability test for flies with a loss of function PINK1 gene (PINK1) and wild-type (WT) flies. Vitamin C was added to levodopa to prevent oxidation.

    Techniques Used:

    36) Product Images from "MFG-E8-derived Peptide MSP68 is a Cytoskeletal Immunomodulator of Neutrophils that Inhibits Rac1"

    Article Title: MFG-E8-derived Peptide MSP68 is a Cytoskeletal Immunomodulator of Neutrophils that Inhibits Rac1

    Journal: The Journal of surgical research

    doi: 10.1016/j.jss.2016.08.098

    A. Rac1 signaling is increased in cells activated for 15 seconds with f -MLP. This signaling is attenuated by pretreatment with MSP68. BMDNs were collected from C57BL/6 mice and treated with PBS, f -MLP, or MSP68 then f -MLP. Cells were lysed and activated Rac1 ELISA was performed. Mean optical densities at 490 nm were assessed. Data are expressed as mean±SEM and compared by one-way ANOVA and Student-Newman-Keuls test, obtained from three independent experiments [PBS (n=6 mice), f -MLP (n=8 mice), MSP68+ f -MLP (n=8 mice)]. *P
    Figure Legend Snippet: A. Rac1 signaling is increased in cells activated for 15 seconds with f -MLP. This signaling is attenuated by pretreatment with MSP68. BMDNs were collected from C57BL/6 mice and treated with PBS, f -MLP, or MSP68 then f -MLP. Cells were lysed and activated Rac1 ELISA was performed. Mean optical densities at 490 nm were assessed. Data are expressed as mean±SEM and compared by one-way ANOVA and Student-Newman-Keuls test, obtained from three independent experiments [PBS (n=6 mice), f -MLP (n=8 mice), MSP68+ f -MLP (n=8 mice)]. *P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay

    A. Expression of phosphorylated P38 after activation if f -MLP with or without MSP68 treatment. BMDNs were collected from C57BL/6 mice and treated with PBS, f -MLP, or MSP68 then f -MLP. Cells were lysed and a western blot was carried out using phosphorylated P38 primary antibody. Data are expressed as mean±SEM and compared by one-way ANOVA and StudentNewman-Keuls test, obtained from three independent experiments [PBS (n=6 mice), f -MLP (n=8 mice), MSP68+ f -MLP (n=8 mice)]. *P
    Figure Legend Snippet: A. Expression of phosphorylated P38 after activation if f -MLP with or without MSP68 treatment. BMDNs were collected from C57BL/6 mice and treated with PBS, f -MLP, or MSP68 then f -MLP. Cells were lysed and a western blot was carried out using phosphorylated P38 primary antibody. Data are expressed as mean±SEM and compared by one-way ANOVA and StudentNewman-Keuls test, obtained from three independent experiments [PBS (n=6 mice), f -MLP (n=8 mice), MSP68+ f -MLP (n=8 mice)]. *P

    Techniques Used: Expressing, Activation Assay, Mouse Assay, Western Blot

    Actin polymerization at 1000×. Phalloidin-Alexafluor488 labeled F-Actin in green. Propidium Iodide labeled nucleus in red. Male C57BL/6 were sacrificed and live bone marrow derived neutrophils were collected. These neutrophils were treated with PBS, f -MLP, or MSP68 followed by f -MLP. Cells stained with Alexafluor488 Phalloidin or Propidium Iodide. Mean fluorescence intensities at 488 nm from low powered fields were measured. Data are expressed as mean±SEM and compared by one-way ANOVA and Student-Newman-Keuls test (n=10,000 events/group). *P
    Figure Legend Snippet: Actin polymerization at 1000×. Phalloidin-Alexafluor488 labeled F-Actin in green. Propidium Iodide labeled nucleus in red. Male C57BL/6 were sacrificed and live bone marrow derived neutrophils were collected. These neutrophils were treated with PBS, f -MLP, or MSP68 followed by f -MLP. Cells stained with Alexafluor488 Phalloidin or Propidium Iodide. Mean fluorescence intensities at 488 nm from low powered fields were measured. Data are expressed as mean±SEM and compared by one-way ANOVA and Student-Newman-Keuls test (n=10,000 events/group). *P

    Techniques Used: Labeling, Derivative Assay, Staining, Fluorescence

    37) Product Images from "Practical considerations for improved reliability and precision during compound specific analysis of δ15N in amino acids using a single combined oxidation-reduction reactor"

    Article Title: Practical considerations for improved reliability and precision during compound specific analysis of δ15N in amino acids using a single combined oxidation-reduction reactor

    Journal: bioRxiv

    doi: 10.1101/638098

    δ 15 N values for glutamic acid (Glu), phenylalanine (Phe), Glu-Phe, and trophic position (TP) for both the method presented here and the previously used method ( Svensson et al. 2016 ). Within the boxplot, open squares represent the mean, lines represent the median, boxes represent the upper and lower quartiles, and whiskers represent the 1.5 quartile ranges. Any black squares outside of the whiskers are outliers.
    Figure Legend Snippet: δ 15 N values for glutamic acid (Glu), phenylalanine (Phe), Glu-Phe, and trophic position (TP) for both the method presented here and the previously used method ( Svensson et al. 2016 ). Within the boxplot, open squares represent the mean, lines represent the median, boxes represent the upper and lower quartiles, and whiskers represent the 1.5 quartile ranges. Any black squares outside of the whiskers are outliers.

    Techniques Used:

    38) Product Images from "Biphasic Effects of α-Asarone on Immobility in the Tail Suspension Test: Evidence for the Involvement of the Noradrenergic and Serotonergic Systems in Its Antidepressant-Like Activity"

    Article Title: Biphasic Effects of α-Asarone on Immobility in the Tail Suspension Test: Evidence for the Involvement of the Noradrenergic and Serotonergic Systems in Its Antidepressant-Like Activity

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2016.00072

    Involvement of noradrenergic system in the antidepressant-like activity of α-asarone. Effect of pre-treatment of mice with (A) AMPT (100 mg/kg, i.p., a catecholamine synthesis inhibitor) or (B) prazosin (1 mg/kg, i.p., an α 1 -adrenoceptor antagonist) or (C) yohimbine (1 mg/kg, i.p., an α 2 -adrenoceptor antagonist) on α-asarone (20 mg/kg, i.p.)- induced anti-immobility in the TST. Values are expressed as mean ± SEM ( n = 8). The immobility time was analyzed using two-way ANOVA followed by post hoc Bonferroni test. ∗∗ p
    Figure Legend Snippet: Involvement of noradrenergic system in the antidepressant-like activity of α-asarone. Effect of pre-treatment of mice with (A) AMPT (100 mg/kg, i.p., a catecholamine synthesis inhibitor) or (B) prazosin (1 mg/kg, i.p., an α 1 -adrenoceptor antagonist) or (C) yohimbine (1 mg/kg, i.p., an α 2 -adrenoceptor antagonist) on α-asarone (20 mg/kg, i.p.)- induced anti-immobility in the TST. Values are expressed as mean ± SEM ( n = 8). The immobility time was analyzed using two-way ANOVA followed by post hoc Bonferroni test. ∗∗ p

    Techniques Used: Activity Assay, Mouse Assay

    39) Product Images from "Cholesterol Is Required for Surface Transport of Influenza Virus Hemagglutinin "

    Article Title: Cholesterol Is Required for Surface Transport of Influenza Virus Hemagglutinin

    Journal: The Journal of Cell Biology

    doi:

    Removal of cholesterol from MDCK cells affects TGN–apical surface transport of influenza virus HA, but not basolateral VSV G transport. ( A and B ) Representative example of a TGN-to-surface transport. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, viral proteins ( A , HA ; B , VSV G ) were chased to the cell surface for different times at 37°C. Arrival of HA on the apical surface was detected by trypsin treatment. Basolateral arrival of VSV G was detected by surface immunoprecipitation. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ( C and D ) Averaged data of five independent experiments (see Fig. 4 ).
    Figure Legend Snippet: Removal of cholesterol from MDCK cells affects TGN–apical surface transport of influenza virus HA, but not basolateral VSV G transport. ( A and B ) Representative example of a TGN-to-surface transport. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, viral proteins ( A , HA ; B , VSV G ) were chased to the cell surface for different times at 37°C. Arrival of HA on the apical surface was detected by trypsin treatment. Basolateral arrival of VSV G was detected by surface immunoprecipitation. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ( C and D ) Averaged data of five independent experiments (see Fig. 4 ).

    Techniques Used: Labeling, Blocking Assay, Immunoprecipitation

    Removal of cholesterol from MDCK cells leads to missorting of gp-80. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling for 15 min with [ 35 S]methionine and a TGN block at 19.5°C, gp-80 was allowed to be secreted for 20 and 40 min at 37°C, and was recovered from apical and basolateral media, as well as from the cells by immunoprecipitation. The immunoprecipitates were analyzed by nonreducing SDS-PAGE, followed by quantitation with a PhosphorImager. ( A ) Untreated control cells. ( B ) Cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ▪, gp-80 secreted apically; □, gp-80, secreted basolaterally.
    Figure Legend Snippet: Removal of cholesterol from MDCK cells leads to missorting of gp-80. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling for 15 min with [ 35 S]methionine and a TGN block at 19.5°C, gp-80 was allowed to be secreted for 20 and 40 min at 37°C, and was recovered from apical and basolateral media, as well as from the cells by immunoprecipitation. The immunoprecipitates were analyzed by nonreducing SDS-PAGE, followed by quantitation with a PhosphorImager. ( A ) Untreated control cells. ( B ) Cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ▪, gp-80 secreted apically; □, gp-80, secreted basolaterally.

    Techniques Used: Labeling, Blocking Assay, Immunoprecipitation, SDS Page, Quantitation Assay

    Treatment of BHK and MDCK cells with methyl- β-cyclodextrin leads to increased Triton X-100 solubility of influenza virus HA. BHK cells ( A ) and filter-grown MDCK cells ( B ) were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. The cells then were extracted on ice with 1% Triton X-100, followed by centrifugation at 120,000 g to obtain detergent-soluble and -insoluble fractions. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin.
    Figure Legend Snippet: Treatment of BHK and MDCK cells with methyl- β-cyclodextrin leads to increased Triton X-100 solubility of influenza virus HA. BHK cells ( A ) and filter-grown MDCK cells ( B ) were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. The cells then were extracted on ice with 1% Triton X-100, followed by centrifugation at 120,000 g to obtain detergent-soluble and -insoluble fractions. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin.

    Techniques Used: Solubility, Centrifugation

    Treatment of BHK cells and of filter-grown MDCK cells with methyl-β-cyclodextrin. BHK cells were left untreated ( A ), or were grown in the presence of lovastatin/mevalonate and then treated for 30 min at 37°C with 10 mM methyl-β-cyclodextrin ( B ) as detailed in Materials and Methods, and then were stained with filipin. Digital images were acquired using 5 ( A ) and 20 ( B ) integration frames, respectively. Filter-grown MDCK cells were left untreated ( C ), or were grown in the presence of lovastatin/mevalonate and then treated for 60 min at 37°C with 10 mM methyl-β-cyclodextrin ( D ) as detailed in Materials and Methods. X–Z confocal views of cells labeled with a monoclonal antibody directed against a basolateral 58-kD protein. Bars, 10 μm.
    Figure Legend Snippet: Treatment of BHK cells and of filter-grown MDCK cells with methyl-β-cyclodextrin. BHK cells were left untreated ( A ), or were grown in the presence of lovastatin/mevalonate and then treated for 30 min at 37°C with 10 mM methyl-β-cyclodextrin ( B ) as detailed in Materials and Methods, and then were stained with filipin. Digital images were acquired using 5 ( A ) and 20 ( B ) integration frames, respectively. Filter-grown MDCK cells were left untreated ( C ), or were grown in the presence of lovastatin/mevalonate and then treated for 60 min at 37°C with 10 mM methyl-β-cyclodextrin ( D ) as detailed in Materials and Methods. X–Z confocal views of cells labeled with a monoclonal antibody directed against a basolateral 58-kD protein. Bars, 10 μm.

    Techniques Used: Staining, Labeling

    Removal of cholesterol does not affect ER-to-Golgi transport. BHK cells ( A and B ) and filter-grown MDCK cells ( C and D ) were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine, viral proteins ( A and C , HA ; B and D , VSV G ) were chased for different times at 37°C. Arrival in the Golgi complex was monitored by acquirement of resistance to endoglycosidase H digestion. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin.
    Figure Legend Snippet: Removal of cholesterol does not affect ER-to-Golgi transport. BHK cells ( A and B ) and filter-grown MDCK cells ( C and D ) were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine, viral proteins ( A and C , HA ; B and D , VSV G ) were chased for different times at 37°C. Arrival in the Golgi complex was monitored by acquirement of resistance to endoglycosidase H digestion. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin.

    Techniques Used: Labeling

    Removal of cholesterol from BHK cells affects TGN-to-surface transport of influenza virus HA, but not of VSV G. ( A and B ) Representative example of a TGN-to-surface transport. BHK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, viral proteins ( A , HA ; B , VSV G ) were chased to the cell surface for different times at 37°C. Arrival on the surface was detected by trypsin treatment ( HA ) or by surface immunoprecipitation ( VSV G ), respectively. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ( C and D ) Averaged data of four independent experiments. Relative transport indices for HA ( C ) and VSV G ( D ) after 15 and 30 min of incubation at 37°C were calculated from four experiments. Transport in cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin was expressed relative to the transport in untreated control cells at the respective time.
    Figure Legend Snippet: Removal of cholesterol from BHK cells affects TGN-to-surface transport of influenza virus HA, but not of VSV G. ( A and B ) Representative example of a TGN-to-surface transport. BHK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, viral proteins ( A , HA ; B , VSV G ) were chased to the cell surface for different times at 37°C. Arrival on the surface was detected by trypsin treatment ( HA ) or by surface immunoprecipitation ( VSV G ), respectively. ▪, untreated control cells; □, cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. ( C and D ) Averaged data of four independent experiments. Relative transport indices for HA ( C ) and VSV G ( D ) after 15 and 30 min of incubation at 37°C were calculated from four experiments. Transport in cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin was expressed relative to the transport in untreated control cells at the respective time.

    Techniques Used: Labeling, Blocking Assay, Immunoprecipitation, Incubation

    Removal of cholesterol from MDCK cells leads to partial missorting of influenza virus HA. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, HA was chased to the cell surface for different times at 37°C. Arrival on the apical or basolateral membrane, respectively, was detected by trypsin treatment. ( A ) untreated control cells; ( B ) cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. HA1 and HA2 are the products generated from HA by trypsin cleavage. The duration of transport (in minutes) is indicated above the lanes.
    Figure Legend Snippet: Removal of cholesterol from MDCK cells leads to partial missorting of influenza virus HA. Filter-grown MDCK cells were treated with lovastatin/mevalonate and methyl-β-cyclodextrin as detailed in Materials and Methods. After pulse labeling with [ 35 S]methionine and a TGN block at 19.5°C, HA was chased to the cell surface for different times at 37°C. Arrival on the apical or basolateral membrane, respectively, was detected by trypsin treatment. ( A ) untreated control cells; ( B ) cells treated with lovastatin/mevalonate and methyl-β-cyclodextrin. HA1 and HA2 are the products generated from HA by trypsin cleavage. The duration of transport (in minutes) is indicated above the lanes.

    Techniques Used: Labeling, Blocking Assay, Generated

    Treatment of BHK and MDCK cells with methyl- β-cyclodextrin leads to efficient removal of cellular cholesterol. BHK cells ( A ) and filter-grown MDCK cells ( B ) were labeled with [ 3 H]cholesterol in the absence (−) or presence (+) of lovastatin/ mevalonate, and radiolabeled cholesterol was allowed to equilibrate with the nonradioactive pool of cholesterol. The cells then were treated for 30 min ( BHK ) or 60 min ( MDCK ) at 37°C with 10 mM methyl-β-cyclodextrin in infection medium (+) or with infection medium only (−). Radioactivity released into the medium and remaining in the cells was determined. Data were obtained from two independent experiments with samples done in duplicate.
    Figure Legend Snippet: Treatment of BHK and MDCK cells with methyl- β-cyclodextrin leads to efficient removal of cellular cholesterol. BHK cells ( A ) and filter-grown MDCK cells ( B ) were labeled with [ 3 H]cholesterol in the absence (−) or presence (+) of lovastatin/ mevalonate, and radiolabeled cholesterol was allowed to equilibrate with the nonradioactive pool of cholesterol. The cells then were treated for 30 min ( BHK ) or 60 min ( MDCK ) at 37°C with 10 mM methyl-β-cyclodextrin in infection medium (+) or with infection medium only (−). Radioactivity released into the medium and remaining in the cells was determined. Data were obtained from two independent experiments with samples done in duplicate.

    Techniques Used: Labeling, Infection, Radioactivity

    40) Product Images from "Functional analysis of the involvement of apurinic/apyrimidinic endonuclease 1 in the resistance to melphalan in multiple myeloma"

    Article Title: Functional analysis of the involvement of apurinic/apyrimidinic endonuclease 1 in the resistance to melphalan in multiple myeloma

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-14-11

    DNA repair activity of APE1 plays a critical role in melphalan resistance of MM cells. (A) The difference in DNA repair activity of APE1 between RPMI-8226/LR5 and RPMI-8226 cells was analyzed using AP site incision assay. The results indicated that the RPMI-8226/LR5 cells possess a higher AP endonuclease activity than its parental line. (B) The overall DNA repair activity for single DNA strand breaks was assayed by the alkaline comet assay. Cell suspensions from both RPMI-8226/LR5 and RPMI-8226 cells were treated with 15 μM melphalan on ice for 20 min, then the comet assay was performed immediately or after a 30 min repair in culture medium in a 37°C incubator. Representative images are shown.
    Figure Legend Snippet: DNA repair activity of APE1 plays a critical role in melphalan resistance of MM cells. (A) The difference in DNA repair activity of APE1 between RPMI-8226/LR5 and RPMI-8226 cells was analyzed using AP site incision assay. The results indicated that the RPMI-8226/LR5 cells possess a higher AP endonuclease activity than its parental line. (B) The overall DNA repair activity for single DNA strand breaks was assayed by the alkaline comet assay. Cell suspensions from both RPMI-8226/LR5 and RPMI-8226 cells were treated with 15 μM melphalan on ice for 20 min, then the comet assay was performed immediately or after a 30 min repair in culture medium in a 37°C incubator. Representative images are shown.

    Techniques Used: Activity Assay, Alkaline Single Cell Gel Electrophoresis, Single Cell Gel Electrophoresis

    APE1 responds to melphalan treatment. Both mRNA and protein levels were elevated after melphalan treatment in a time course in RPMI-8226 cells. Quantitative PCR results are shown as a bar graph in (A) and representative Western blot images are shown in (B) . In addition, expression of APE1 mRNA (C) and protein (D) levels increased in a melphalan dose dependent manner. (E) APE1 protein level alterations in melphalan-resistant line RPMI-8226/LR5 were tested by Western blot at different time points post 15 μM melphalan treatment (left panel) or at 24 hours post various doses of melphalan treatment (right panel). All quantitative RT-PCR results were statistically processed from three independent experiments, and the blot is a representative of three independent Western blots. Stars (*) represent that the difference between the indicated group and DMSO (vehicle) treated RPMI-8226 is statistically significant ( p
    Figure Legend Snippet: APE1 responds to melphalan treatment. Both mRNA and protein levels were elevated after melphalan treatment in a time course in RPMI-8226 cells. Quantitative PCR results are shown as a bar graph in (A) and representative Western blot images are shown in (B) . In addition, expression of APE1 mRNA (C) and protein (D) levels increased in a melphalan dose dependent manner. (E) APE1 protein level alterations in melphalan-resistant line RPMI-8226/LR5 were tested by Western blot at different time points post 15 μM melphalan treatment (left panel) or at 24 hours post various doses of melphalan treatment (right panel). All quantitative RT-PCR results were statistically processed from three independent experiments, and the blot is a representative of three independent Western blots. Stars (*) represent that the difference between the indicated group and DMSO (vehicle) treated RPMI-8226 is statistically significant ( p

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

    Manipulation of APE1 affects cell resistance to melphalan. (A) APE1 protein levels at 48 hours post transfection of siRNA or overexpression vector in RPMI-8226 cells were measured using Western blot. Transfection reagent only treated cells were included as a control. (B) Cell survival after 24, 48 and 72 hours post 15 μM melphalan treatment in all three groups was measured using CCK-8 assay. A star (*) represents that the difference between the shAPE1 transfected group and the vehicle alone group is statistically significant ( p
    Figure Legend Snippet: Manipulation of APE1 affects cell resistance to melphalan. (A) APE1 protein levels at 48 hours post transfection of siRNA or overexpression vector in RPMI-8226 cells were measured using Western blot. Transfection reagent only treated cells were included as a control. (B) Cell survival after 24, 48 and 72 hours post 15 μM melphalan treatment in all three groups was measured using CCK-8 assay. A star (*) represents that the difference between the shAPE1 transfected group and the vehicle alone group is statistically significant ( p

    Techniques Used: Transfection, Over Expression, Plasmid Preparation, Western Blot, CCK-8 Assay

    Differential involvement of various APE1 functions in melphalan-resistanct MM cells. (A) The protein expression levels of APE1 mutants were assayed by Western blot and the results indicated that the exogenous protein levels were comparable in APE1 WT , APE1 H309N , APE1 C65S and APE1 K6R/K7R cell lines. (B) The AP endonuclease activities of the whole cell extracts from APE1 shRNA and exogenous APE1-expressing cells were measured by oligo incision assay. The representative image out of three experimental repeats was shown. (C) Cell survival of different APE1 mutant-transfected groups was measured by CCK-8 assay after various doses of melphalan treatment. All results were from three independent experiments and * represents that the difference between the indicated group and the APE1 WT group is statistically significant ( p
    Figure Legend Snippet: Differential involvement of various APE1 functions in melphalan-resistanct MM cells. (A) The protein expression levels of APE1 mutants were assayed by Western blot and the results indicated that the exogenous protein levels were comparable in APE1 WT , APE1 H309N , APE1 C65S and APE1 K6R/K7R cell lines. (B) The AP endonuclease activities of the whole cell extracts from APE1 shRNA and exogenous APE1-expressing cells were measured by oligo incision assay. The representative image out of three experimental repeats was shown. (C) Cell survival of different APE1 mutant-transfected groups was measured by CCK-8 assay after various doses of melphalan treatment. All results were from three independent experiments and * represents that the difference between the indicated group and the APE1 WT group is statistically significant ( p

    Techniques Used: Expressing, Western Blot, shRNA, Mutagenesis, Transfection, CCK-8 Assay

    MDR1 expression is aberrant in APE1 acetylation site mutant expressing MM cells. (A) At 2 hours after melphalan treatment, APE1 was enriched by pulldown and blotted with anti-lysine acetylation antibody. (B) The Western blot showed expression levels of MDR1 in RPMI-8226/LR5, U266/LR6 and their parental cell lines RPMI-8226 and U266. The representative blots showed that melphalan resistant cells have higher expression levels of MDR1. (C) MDR1 protein expression was detected by Western blot at 48 hours after APE1 siRNA or pcDNA-APE1 was transfected into RPMI-8226 cells. MDR1 expression was downregulated after knockdown of APE1 in RPMI-8226 cells, and MDR1 was upregulated after overexpression of APE1. In addition, APE1 WT or APE1 K6R/K7R was transfected 24 hours after APE1 shRNA infection. 48 hours later, MDR1 levels were detected by Western blot. (D) MDR1 siRNA was applied to the pcDNA-APE1 transfected RPMI-8226 cells at 48 hours post 15 μM melphalan treatment, and cell viability was measured by a CCK-8 kit. The results were from three independent experiments.
    Figure Legend Snippet: MDR1 expression is aberrant in APE1 acetylation site mutant expressing MM cells. (A) At 2 hours after melphalan treatment, APE1 was enriched by pulldown and blotted with anti-lysine acetylation antibody. (B) The Western blot showed expression levels of MDR1 in RPMI-8226/LR5, U266/LR6 and their parental cell lines RPMI-8226 and U266. The representative blots showed that melphalan resistant cells have higher expression levels of MDR1. (C) MDR1 protein expression was detected by Western blot at 48 hours after APE1 siRNA or pcDNA-APE1 was transfected into RPMI-8226 cells. MDR1 expression was downregulated after knockdown of APE1 in RPMI-8226 cells, and MDR1 was upregulated after overexpression of APE1. In addition, APE1 WT or APE1 K6R/K7R was transfected 24 hours after APE1 shRNA infection. 48 hours later, MDR1 levels were detected by Western blot. (D) MDR1 siRNA was applied to the pcDNA-APE1 transfected RPMI-8226 cells at 48 hours post 15 μM melphalan treatment, and cell viability was measured by a CCK-8 kit. The results were from three independent experiments.

    Techniques Used: Expressing, Mutagenesis, Western Blot, Transfection, Over Expression, shRNA, Infection, CCK-8 Assay

    APE1 overexpression in melphalan-resistant MM cell lines RPMI-8226/LR5 and U266/LR6. (A) CCK-8 assay indicated that RPMI-8226/LR5 and U266/LR6 show more resistance to melphalan than their parental cell lines RPMI-8226 and U266. All cell lines were treated with three different doses of melphalan for 48 hours then cell viability was assayed using the CCK-8 assay. Results are shown as mean ± SD and were from three separate experiments. The significance was analyzed by Student t test. Stars (*) represent that the differences between RPMI-8226/LR5 and RPMI-8226 or between U266/LR6 and U266 are statistically significant ( p
    Figure Legend Snippet: APE1 overexpression in melphalan-resistant MM cell lines RPMI-8226/LR5 and U266/LR6. (A) CCK-8 assay indicated that RPMI-8226/LR5 and U266/LR6 show more resistance to melphalan than their parental cell lines RPMI-8226 and U266. All cell lines were treated with three different doses of melphalan for 48 hours then cell viability was assayed using the CCK-8 assay. Results are shown as mean ± SD and were from three separate experiments. The significance was analyzed by Student t test. Stars (*) represent that the differences between RPMI-8226/LR5 and RPMI-8226 or between U266/LR6 and U266 are statistically significant ( p

    Techniques Used: Over Expression, CCK-8 Assay

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  • 97
    Millipore calpain inhibitor mdl 28170
    Effect of MDL 28170, a potent inhibitor of <t>calpain</t> I and II, on the growth rate of Leishmania amazonensis . The growth pattern of L. amazonensis was followed for parasites cultivated at 26 °C in the absence (control) or presence of <t>MDL</t> 28170 at concentrations ranging from 15 μM to 30 μM. The inhibitor was added to the cultures at 0 h and the cells were counted daily. Data shown are the mean ± standard deviation (S.D.) of three independent experiments performed in triplicate. The bars indicate the S.D.
    Calpain Inhibitor Mdl 28170, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore dota aoc d phe d lys6 gnrh
    The competitive binding curves of the <t>GnRH</t> peptides. The IC 50 values of <t>DOTA-Aoc-D-Phe-(D-Lys</t> 6 -GnRH), DOTA-β-Ala-D-Phe-(D-Lys 6 -GnRH) and DOTA-Aun-D-Phe-(D-Lys 6 -GnRH) were 6.6 ± 0.1, 13.5 ± 3.9 and 24.6 ± 2.5 nM, respectively.
    Dota Aoc D Phe D Lys6 Gnrh, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore batimastat
    Rescue of apoptosis by protease inhibitors in normal and CCh fibroblasts promoted by PTX3 knockdown. Both normal and CCh fibroblasts were cultured in DMEM with 0.5% FBS for 48 hours and transfected with PTX3 siRNA for another 48 hours, while IL-1β was added for the last 24 hours. ( A ) Phase contrast microscopy of normal ( a – e ) and CCh ( f – j ) fibroblasts showed that PTX3 siRNA ( b , g ) caused more detached round cells than scRNA ( a , f ). However, such changes were abolished by pretreatment with three protease inhibitors for 1 hour (Aprotinin [ c , h ]; <t>Batimastat,</t> [ d , i ]; and NNGH, [ e , j ], respectively). Scale bar = 100 μm. ( B ) The extent of cell apoptosis in cell lysates ( top ) and necrosis in culture media ( bottom ) were also significantly increased by PTX3 siRNA treatment (count percentage of apoptosis or necrosis cell numbers in 2000 total cells each experiment, * P
    Batimastat, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of MDL 28170, a potent inhibitor of calpain I and II, on the growth rate of Leishmania amazonensis . The growth pattern of L. amazonensis was followed for parasites cultivated at 26 °C in the absence (control) or presence of MDL 28170 at concentrations ranging from 15 μM to 30 μM. The inhibitor was added to the cultures at 0 h and the cells were counted daily. Data shown are the mean ± standard deviation (S.D.) of three independent experiments performed in triplicate. The bars indicate the S.D.

    Journal: International Journal of Antimicrobial Agents

    Article Title: Antileishmanial activity of MDL 28170, a potent calpain inhibitor

    doi: 10.1016/j.ijantimicag.2006.03.021

    Figure Lengend Snippet: Effect of MDL 28170, a potent inhibitor of calpain I and II, on the growth rate of Leishmania amazonensis . The growth pattern of L. amazonensis was followed for parasites cultivated at 26 °C in the absence (control) or presence of MDL 28170 at concentrations ranging from 15 μM to 30 μM. The inhibitor was added to the cultures at 0 h and the cells were counted daily. Data shown are the mean ± standard deviation (S.D.) of three independent experiments performed in triplicate. The bars indicate the S.D.

    Article Snippet: 2.1 Chemicals The calpain inhibitor MDL 28170 (carbobenzoxy-valyl-phenylalanial; Z-Val-Phe-CHO) was purchased from Calbiochem (San Diego, CA).

    Techniques: Standard Deviation

    The competitive binding curves of the GnRH peptides. The IC 50 values of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH), DOTA-β-Ala-D-Phe-(D-Lys 6 -GnRH) and DOTA-Aun-D-Phe-(D-Lys 6 -GnRH) were 6.6 ± 0.1, 13.5 ± 3.9 and 24.6 ± 2.5 nM, respectively.

    Journal: Bioorganic & medicinal chemistry letters

    Article Title: Evaluation of novel 111In-labeled gonadotropin-releasing hormone peptides for human prostate cancer imaging

    doi: 10.1016/j.bmcl.2017.09.004

    Figure Lengend Snippet: The competitive binding curves of the GnRH peptides. The IC 50 values of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH), DOTA-β-Ala-D-Phe-(D-Lys 6 -GnRH) and DOTA-Aun-D-Phe-(D-Lys 6 -GnRH) were 6.6 ± 0.1, 13.5 ± 3.9 and 24.6 ± 2.5 nM, respectively.

    Article Snippet: The GnRH receptor binding affinities of DOTA-βAla-D-Phe-(D-Lys6 -GnRH), DOTA-Aoc-D-Phe-(D-Lys6 -GnRH) and DOTA-Aun-D-Phe-(D-Lys6 -GnRH) were determined using Millipore ChemiScreen™ human GnRH receptor membrane preparations.

    Techniques: Binding Assay

    Radioactive HPLC profile of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) (A) and UV HPLC profile of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) (B). The retention times of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) and DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) were 16.6 and 13.6 min, respectively. Binding of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) on human GnRH receptor membrane preparations with (white) or without (black) the presence of 1 μM of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH).

    Journal: Bioorganic & medicinal chemistry letters

    Article Title: Evaluation of novel 111In-labeled gonadotropin-releasing hormone peptides for human prostate cancer imaging

    doi: 10.1016/j.bmcl.2017.09.004

    Figure Lengend Snippet: Radioactive HPLC profile of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) (A) and UV HPLC profile of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) (B). The retention times of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) and DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) were 16.6 and 13.6 min, respectively. Binding of 111 In-DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH) on human GnRH receptor membrane preparations with (white) or without (black) the presence of 1 μM of DOTA-Aoc-D-Phe-(D-Lys 6 -GnRH).

    Article Snippet: The GnRH receptor binding affinities of DOTA-βAla-D-Phe-(D-Lys6 -GnRH), DOTA-Aoc-D-Phe-(D-Lys6 -GnRH) and DOTA-Aun-D-Phe-(D-Lys6 -GnRH) were determined using Millipore ChemiScreen™ human GnRH receptor membrane preparations.

    Techniques: High Performance Liquid Chromatography, Binding Assay

    Rescue of apoptosis by protease inhibitors in normal and CCh fibroblasts promoted by PTX3 knockdown. Both normal and CCh fibroblasts were cultured in DMEM with 0.5% FBS for 48 hours and transfected with PTX3 siRNA for another 48 hours, while IL-1β was added for the last 24 hours. ( A ) Phase contrast microscopy of normal ( a – e ) and CCh ( f – j ) fibroblasts showed that PTX3 siRNA ( b , g ) caused more detached round cells than scRNA ( a , f ). However, such changes were abolished by pretreatment with three protease inhibitors for 1 hour (Aprotinin [ c , h ]; Batimastat, [ d , i ]; and NNGH, [ e , j ], respectively). Scale bar = 100 μm. ( B ) The extent of cell apoptosis in cell lysates ( top ) and necrosis in culture media ( bottom ) were also significantly increased by PTX3 siRNA treatment (count percentage of apoptosis or necrosis cell numbers in 2000 total cells each experiment, * P

    Journal: Investigative Ophthalmology & Visual Science

    Article Title: PTX3 Controls Activation of Matrix Metalloproteinase 1 and Apoptosis in Conjunctivochalasis Fibroblasts

    doi: 10.1167/iovs.11-9103

    Figure Lengend Snippet: Rescue of apoptosis by protease inhibitors in normal and CCh fibroblasts promoted by PTX3 knockdown. Both normal and CCh fibroblasts were cultured in DMEM with 0.5% FBS for 48 hours and transfected with PTX3 siRNA for another 48 hours, while IL-1β was added for the last 24 hours. ( A ) Phase contrast microscopy of normal ( a – e ) and CCh ( f – j ) fibroblasts showed that PTX3 siRNA ( b , g ) caused more detached round cells than scRNA ( a , f ). However, such changes were abolished by pretreatment with three protease inhibitors for 1 hour (Aprotinin [ c , h ]; Batimastat, [ d , i ]; and NNGH, [ e , j ], respectively). Scale bar = 100 μm. ( B ) The extent of cell apoptosis in cell lysates ( top ) and necrosis in culture media ( bottom ) were also significantly increased by PTX3 siRNA treatment (count percentage of apoptosis or necrosis cell numbers in 2000 total cells each experiment, * P

    Article Snippet: Batimastat, an inhibitor of both MMP1 and MMP3, and centrifugal filter units were purchased from EMD Millipore (Billerica, MA).

    Techniques: Cell Culture, Transfection, Microscopy

    BPA and NP induce shedding of TNF-α and Neuregulin in cultured rat Sertoli cells. A) The amount of TNF-α in the culture medium of primary rats Sertoli cells significantly increases after 6 h of incubation with 10 µM BPA or NP, but remains similar to control levels in the presence of 10 µM BB-94, a general metalloprotease inhibitor. The picture shows immunofluorescence against ADAM17 (green) in rat primary Sertoli cells; cell nuclei are in red stained with Propidium iodide (IP). B) The amplicon of NGR-β1 is detected only in transfected rat primary Sertoli cells with the (AP)-NGR-β1 vector. C) Non-transfected primary Sertoli cells show no AP activity, at any of the tested concentrations. D) BPA and NP (E), induce shedding of AP activity in culture medium after 24 hours of stimulation, which is prevented by BB-94. * p

    Journal: PLoS ONE

    Article Title: A Mechanism of Male Germ Cell Apoptosis Induced by Bisphenol-A and Nonylphenol Involving ADAM17 and p38 MAPK Activation

    doi: 10.1371/journal.pone.0113793

    Figure Lengend Snippet: BPA and NP induce shedding of TNF-α and Neuregulin in cultured rat Sertoli cells. A) The amount of TNF-α in the culture medium of primary rats Sertoli cells significantly increases after 6 h of incubation with 10 µM BPA or NP, but remains similar to control levels in the presence of 10 µM BB-94, a general metalloprotease inhibitor. The picture shows immunofluorescence against ADAM17 (green) in rat primary Sertoli cells; cell nuclei are in red stained with Propidium iodide (IP). B) The amplicon of NGR-β1 is detected only in transfected rat primary Sertoli cells with the (AP)-NGR-β1 vector. C) Non-transfected primary Sertoli cells show no AP activity, at any of the tested concentrations. D) BPA and NP (E), induce shedding of AP activity in culture medium after 24 hours of stimulation, which is prevented by BB-94. * p

    Article Snippet: The p38 MAPK inhibitor, PD169316 (513030), and a general metalloprotease inhibitor, BB-94 (Batimastat), were obtained from Calbiochem (San Diego, CA, USA).

    Techniques: Cell Culture, Incubation, Immunofluorescence, Staining, Amplification, Transfection, Plasmid Preparation, Activity Assay