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  • 99
    Name:
    TRIzol Reagent
    Description:
    TRIzol Reagent is a complete ready to use reagent for the isolation of high quality total RNA or the simultaneous isolation of RNA DNA and protein from a variety of biological samples This monophasic solution of phenol and guanidine isothiocyanate is designed to isolate separate fractions of RNA DNA and proteins from cell and tissue samples of human animal plant yeast or bacterial origin within one hour Key features of TRIzol Reagent include • Permits the isolation of RNA DNA and protein from the same sample• Offers superior lysis capability even with difficult sample types• Optimized formulations and protocols for tissues cells serum virus and bacteriaReliably purify RNA from multiple sample volumes and sourcesTRIzol Reagent performs well with small quantities of tissue 50 100 mg and cells 5 × 106 as well as with large quantities of tissue 1 g and cells 107 and comes with prototcols for purification from samples of human animal plant or bacterial origin TRIzol Reagent maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization The simplicity of the TRIzol Reagent method allows simultaneous processing of a large number of samples The entire procedure can be completed in 1 hour Total RNA isolated by TRIzol Reagent is free of protein and DNA contamination Formulated for isolation of multiple molecular targetsTRIzol Reagent allows you to perform sequential precipitation of RNA DNA and proteins from a single sample After homogenizing the sample with TRIzol Reagent chloroform is added and the homogenate is allowed to separate into a clear upper aqueous layer containing RNA and interphase and red lower organic layers containing the DNA and proteins RNA is precipitated from the aqueous layer with isopropanol DNA is precipitated from the interphase organic layer with ethanol Protein is precipitated from the phenol ethanol supernatant by isopropanol precipitation The precipitated RNA DNA or protein is washed to remove impurities and then resuspended for use in downstream applications
    Catalog Number:
    15596018
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|RNAi, Epigenetics & Non-Coding RNA Research|RNA Extraction|Total RNA Isolation|Total RNA from Animal Cells & Tissues|Total RNA from Bacterial Samples|Total RNA from Plant Cells|Total RNA from Yeast|miRNA Isolation|miRNA & Non-Coding RNA Analysis
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher tbe
    Non-denaturing agarose gel (1.5%) electrophoresis in <t>TBE</t> with in-gel <t>ethidium</t> bromide staining of RNA isolated from Arabidopsis, wheat and tomato leaves using TRI reagent (SIGMA) (labelled as 1) or the protocol by Oñate-Sánchez and Vicente-Carbajosa [ 4 ] (labelled as 2). In all cases, 500 theoretical nanograms, according to the spectrophotometric quantification, were loaded per lane. Samples were extracted and analysed in triplicates. Starting material per sample was as follows: Arabidopsis – eight leaf discs (7 mm diameter) of five-week-old plants grown in short day conditions; wheat – eight leaf discs (7 mm diameter) of four-week-old plants grown in the glasshouse; tomato – eight leaf discs (7 mm diameter) of five-week-old plants grown in the glasshouse. Samples were resuspended in 30 μL of water in all cases. The RNA ladder is the 0.5-10 Kb RNA ladder (Life Technologies).
    TRIzol Reagent is a complete ready to use reagent for the isolation of high quality total RNA or the simultaneous isolation of RNA DNA and protein from a variety of biological samples This monophasic solution of phenol and guanidine isothiocyanate is designed to isolate separate fractions of RNA DNA and proteins from cell and tissue samples of human animal plant yeast or bacterial origin within one hour Key features of TRIzol Reagent include • Permits the isolation of RNA DNA and protein from the same sample• Offers superior lysis capability even with difficult sample types• Optimized formulations and protocols for tissues cells serum virus and bacteriaReliably purify RNA from multiple sample volumes and sourcesTRIzol Reagent performs well with small quantities of tissue 50 100 mg and cells 5 × 106 as well as with large quantities of tissue 1 g and cells 107 and comes with prototcols for purification from samples of human animal plant or bacterial origin TRIzol Reagent maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization The simplicity of the TRIzol Reagent method allows simultaneous processing of a large number of samples The entire procedure can be completed in 1 hour Total RNA isolated by TRIzol Reagent is free of protein and DNA contamination Formulated for isolation of multiple molecular targetsTRIzol Reagent allows you to perform sequential precipitation of RNA DNA and proteins from a single sample After homogenizing the sample with TRIzol Reagent chloroform is added and the homogenate is allowed to separate into a clear upper aqueous layer containing RNA and interphase and red lower organic layers containing the DNA and proteins RNA is precipitated from the aqueous layer with isopropanol DNA is precipitated from the interphase organic layer with ethanol Protein is precipitated from the phenol ethanol supernatant by isopropanol precipitation The precipitated RNA DNA or protein is washed to remove impurities and then resuspended for use in downstream applications
    https://www.bioz.com/result/tbe/product/Thermo Fisher
    Average 99 stars, based on 11226 article reviews
    Price from $9.99 to $1999.99
    tbe - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "Broad application of a simple and affordable protocol for isolating plant RNA"

    Article Title: Broad application of a simple and affordable protocol for isolating plant RNA

    Journal: BMC Research Notes

    doi: 10.1186/s13104-015-1119-7

    Non-denaturing agarose gel (1.5%) electrophoresis in TBE with in-gel ethidium bromide staining of RNA isolated from Arabidopsis, wheat and tomato leaves using TRI reagent (SIGMA) (labelled as 1) or the protocol by Oñate-Sánchez and Vicente-Carbajosa [ 4 ] (labelled as 2). In all cases, 500 theoretical nanograms, according to the spectrophotometric quantification, were loaded per lane. Samples were extracted and analysed in triplicates. Starting material per sample was as follows: Arabidopsis – eight leaf discs (7 mm diameter) of five-week-old plants grown in short day conditions; wheat – eight leaf discs (7 mm diameter) of four-week-old plants grown in the glasshouse; tomato – eight leaf discs (7 mm diameter) of five-week-old plants grown in the glasshouse. Samples were resuspended in 30 μL of water in all cases. The RNA ladder is the 0.5-10 Kb RNA ladder (Life Technologies).
    Figure Legend Snippet: Non-denaturing agarose gel (1.5%) electrophoresis in TBE with in-gel ethidium bromide staining of RNA isolated from Arabidopsis, wheat and tomato leaves using TRI reagent (SIGMA) (labelled as 1) or the protocol by Oñate-Sánchez and Vicente-Carbajosa [ 4 ] (labelled as 2). In all cases, 500 theoretical nanograms, according to the spectrophotometric quantification, were loaded per lane. Samples were extracted and analysed in triplicates. Starting material per sample was as follows: Arabidopsis – eight leaf discs (7 mm diameter) of five-week-old plants grown in short day conditions; wheat – eight leaf discs (7 mm diameter) of four-week-old plants grown in the glasshouse; tomato – eight leaf discs (7 mm diameter) of five-week-old plants grown in the glasshouse. Samples were resuspended in 30 μL of water in all cases. The RNA ladder is the 0.5-10 Kb RNA ladder (Life Technologies).

    Techniques Used: Agarose Gel Electrophoresis, Electrophoresis, Staining, Isolation

    2) Product Images from "Negative Cellular Effects of Urban Particulate Matter on Human Keratinocytes Are Mediated by P38 MAPK and NF-κB-dependent Expression of TRPV 1"

    Article Title: Negative Cellular Effects of Urban Particulate Matter on Human Keratinocytes Are Mediated by P38 MAPK and NF-κB-dependent Expression of TRPV 1

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19092660

    UPM-induced expression of TRPV 1 is mediated through activation of p38 mitogen-activated protein kinase (MAPK) and NF-κB. HaCaT cells were treated with UPM (200 ppm) and then incubated for two days in the presence of the indicated concentration of MAPK and NF-κB inhibitors. ( A ) Total RNA was isolated from the cells, and the mRNA levels of the indicated genes were measured using real-time quantitative RT-PCR. The expressed results are relative to the untreated cells after normalization against the GAPDH level. * p
    Figure Legend Snippet: UPM-induced expression of TRPV 1 is mediated through activation of p38 mitogen-activated protein kinase (MAPK) and NF-κB. HaCaT cells were treated with UPM (200 ppm) and then incubated for two days in the presence of the indicated concentration of MAPK and NF-κB inhibitors. ( A ) Total RNA was isolated from the cells, and the mRNA levels of the indicated genes were measured using real-time quantitative RT-PCR. The expressed results are relative to the untreated cells after normalization against the GAPDH level. * p

    Techniques Used: Expressing, Activation Assay, Incubation, Concentration Assay, Isolation, Quantitative RT-PCR

    3) Product Images from "A novel mechanism of natural killer cell response to anti-CTLA-4 therapy identified by integrative analysis of mouse and human tumors"

    Article Title: A novel mechanism of natural killer cell response to anti-CTLA-4 therapy identified by integrative analysis of mouse and human tumors

    Journal: bioRxiv

    doi: 10.1101/2020.05.31.125625

    CD28 and CTLA-4 are coexpressed in circulating and tumoral NK cells. A. Flow cytometry for surface expression of CD28 in NK cell lines (NK-92, NKL, YT, KHYG-1). B. qRT-PCR analysis of CD28 expression in a CD28 null line (PANC-1), T cell lines (Jurkat, CEM), NK cell lines (NK-92, NKL, YT, KHYG-1). C. qRT-PCR demonstrating CD28 expression in CD56+ selected ex vivo unstimulated NK cells derived from PBMCs from healthy human donors. D. scRNAseq data demonstrating positive correlation (R 2 = 0.33, p = 0) between CD28 gene expression and CTLA-4 gene expression in human natural killer cells. E. B7.2 (CD86) mRNA expression levels in primary tumors versus paired normal (TCGA). Red asterisk (*) indicates cancer types with significantly overexpressed CD86 in tumor compared to normal tissue. F. Pan-cancer TCGA data demonstrating a negative correlation between pattern 7 weight and B7.2 expression. Significant correlation coefficients are indicated by a red asterisk (*). G. Schematic demonstrating proposed mechanism by which anti-CTLA-4 antibodies may enhance NK cell-mediated tumor clearance.
    Figure Legend Snippet: CD28 and CTLA-4 are coexpressed in circulating and tumoral NK cells. A. Flow cytometry for surface expression of CD28 in NK cell lines (NK-92, NKL, YT, KHYG-1). B. qRT-PCR analysis of CD28 expression in a CD28 null line (PANC-1), T cell lines (Jurkat, CEM), NK cell lines (NK-92, NKL, YT, KHYG-1). C. qRT-PCR demonstrating CD28 expression in CD56+ selected ex vivo unstimulated NK cells derived from PBMCs from healthy human donors. D. scRNAseq data demonstrating positive correlation (R 2 = 0.33, p = 0) between CD28 gene expression and CTLA-4 gene expression in human natural killer cells. E. B7.2 (CD86) mRNA expression levels in primary tumors versus paired normal (TCGA). Red asterisk (*) indicates cancer types with significantly overexpressed CD86 in tumor compared to normal tissue. F. Pan-cancer TCGA data demonstrating a negative correlation between pattern 7 weight and B7.2 expression. Significant correlation coefficients are indicated by a red asterisk (*). G. Schematic demonstrating proposed mechanism by which anti-CTLA-4 antibodies may enhance NK cell-mediated tumor clearance.

    Techniques Used: Flow Cytometry, Expressing, Quantitative RT-PCR, Ex Vivo, Derivative Assay

    CTLA-4 is expressed by both human NK cell lines and healthy human donor-derived NK cells. A. UMAP dimension reduction with cells colored by single-cell gene expression for CTLA-4 and representative immune activation genes in mouse (left) and human (right) intratumoral NK cells. The pattern of CTLA-4 expression is consistent with the reduced ability of scRNA-seq to capture low to moderately expressed genes. B. Flow cytometry for surface expression of CTLA-4 in positive control (Jurkat-CTLA4) and NK cell lines (NK-92, NKL, YT, KHYG-1). C. Quantitative real-time PCR (qrt-PCR) analysis of CTLA-4 expression in a CTLA-4 null line (PANC-1), T cell lines (Jurkat, CEM, HuT78), NK cell lines (NK-92, NKL, YT, KHYG-1). D. Western blot demonstrating CTLA-4 expression in human NK cell lines. E. qrt-PCR demonstrating CTLA-4 expression in CD56+ selected ex vivo unstimulated NK cells derived from healthy human donors. Graphs are representative of 4 donors. F. Western blot of CTLA-4 expression in CD56+ selected ex vivo unstimulated NK cells derived from healthy human donors. Blots are representative of 4 donors.
    Figure Legend Snippet: CTLA-4 is expressed by both human NK cell lines and healthy human donor-derived NK cells. A. UMAP dimension reduction with cells colored by single-cell gene expression for CTLA-4 and representative immune activation genes in mouse (left) and human (right) intratumoral NK cells. The pattern of CTLA-4 expression is consistent with the reduced ability of scRNA-seq to capture low to moderately expressed genes. B. Flow cytometry for surface expression of CTLA-4 in positive control (Jurkat-CTLA4) and NK cell lines (NK-92, NKL, YT, KHYG-1). C. Quantitative real-time PCR (qrt-PCR) analysis of CTLA-4 expression in a CTLA-4 null line (PANC-1), T cell lines (Jurkat, CEM, HuT78), NK cell lines (NK-92, NKL, YT, KHYG-1). D. Western blot demonstrating CTLA-4 expression in human NK cell lines. E. qrt-PCR demonstrating CTLA-4 expression in CD56+ selected ex vivo unstimulated NK cells derived from healthy human donors. Graphs are representative of 4 donors. F. Western blot of CTLA-4 expression in CD56+ selected ex vivo unstimulated NK cells derived from healthy human donors. Blots are representative of 4 donors.

    Techniques Used: Derivative Assay, Expressing, Activation Assay, Flow Cytometry, Positive Control, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Ex Vivo

    4) Product Images from "Molecular cloning and expression analysis of adiponectin and its receptors (AdipoR1 and AdipoR2) in the hypothalamus of the Huoyan goose during different stages of the egg-laying cycle"

    Article Title: Molecular cloning and expression analysis of adiponectin and its receptors (AdipoR1 and AdipoR2) in the hypothalamus of the Huoyan goose during different stages of the egg-laying cycle

    Journal: Reproductive Biology and Endocrinology : RB & E

    doi: 10.1186/s12958-015-0085-1

    Relative expressions of adiponectin, AdipoR1 and AdipoR2 protein in the hypothalamus of Huoyan geese during different stages of the egg-laying cycle. A comparison of adiponectin, AdipoR1 and AdipoR2 protein content were determined by western blotting analysis. Protein band density was analysed with GelQuant software. Beta-Actin was used as the internal control. Upper panels: representative immunoblots. Lower panels: densitometric analysis of adiponectin, AdipoR1 and AdipoR2 proteins relative to actin protein. Values are expressed as the mean ± SEM of arbitrary optical density units. Data were analysed by ANOVA followed by Tamhane’s T2 test post hoc test. Bars with different superscripts are significantly different ( P
    Figure Legend Snippet: Relative expressions of adiponectin, AdipoR1 and AdipoR2 protein in the hypothalamus of Huoyan geese during different stages of the egg-laying cycle. A comparison of adiponectin, AdipoR1 and AdipoR2 protein content were determined by western blotting analysis. Protein band density was analysed with GelQuant software. Beta-Actin was used as the internal control. Upper panels: representative immunoblots. Lower panels: densitometric analysis of adiponectin, AdipoR1 and AdipoR2 proteins relative to actin protein. Values are expressed as the mean ± SEM of arbitrary optical density units. Data were analysed by ANOVA followed by Tamhane’s T2 test post hoc test. Bars with different superscripts are significantly different ( P

    Techniques Used: Western Blot, Software

    Relative expression of adiponectin, AdipoR1 and AdipoR2 mRNA in the hypothalamus of Huoyan geese during different stages of the egg-laying cycle. The expression levels of adiponectin, AdipoR1 and AdipoR2 were normalized to 18S rRNA. The expression levels, calculated by the 2 −ΔΔCt method, are presented as arbitrary units (AU). The presented values are the means ± SEM. The data were analysed by ANOVA followed by Tamhane’s T2 test post hoc test. Bars with different superscripts are significantly different ( P
    Figure Legend Snippet: Relative expression of adiponectin, AdipoR1 and AdipoR2 mRNA in the hypothalamus of Huoyan geese during different stages of the egg-laying cycle. The expression levels of adiponectin, AdipoR1 and AdipoR2 were normalized to 18S rRNA. The expression levels, calculated by the 2 −ΔΔCt method, are presented as arbitrary units (AU). The presented values are the means ± SEM. The data were analysed by ANOVA followed by Tamhane’s T2 test post hoc test. Bars with different superscripts are significantly different ( P

    Techniques Used: Expressing

    5) Product Images from "Global Array-Based Transcriptomics from Minimal Input RNA Utilising an Optimal RNA Isolation Process Combined with SPIA cDNA Probes"

    Article Title: Global Array-Based Transcriptomics from Minimal Input RNA Utilising an Optimal RNA Isolation Process Combined with SPIA cDNA Probes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0017625

    Comparison of experimental workflows for optimised total RNA isolation from minimal cell numbers. ( A ) Titrated HUVEC samples were processed for RNA isolation through Process A or B to establish the optimal workflow for RNA yield from minimal cell numbers. ( B ) HUVECs were titrated over a range of 3000 - 200 cells per tube and split in to 6 aliquots for cell lysis and RNA isolation. Triplicate samples were processed using either process A or B. All samples were then quantified by reverse transcription of the entire yielded RNA and real-time quantitative PCR using SybrGreen probe and β-actin primers against a standard curve of known HUVEC RNA input. Error bars = standard deviation.
    Figure Legend Snippet: Comparison of experimental workflows for optimised total RNA isolation from minimal cell numbers. ( A ) Titrated HUVEC samples were processed for RNA isolation through Process A or B to establish the optimal workflow for RNA yield from minimal cell numbers. ( B ) HUVECs were titrated over a range of 3000 - 200 cells per tube and split in to 6 aliquots for cell lysis and RNA isolation. Triplicate samples were processed using either process A or B. All samples were then quantified by reverse transcription of the entire yielded RNA and real-time quantitative PCR using SybrGreen probe and β-actin primers against a standard curve of known HUVEC RNA input. Error bars = standard deviation.

    Techniques Used: Isolation, Lysis, Real-time Polymerase Chain Reaction, Standard Deviation

    Assignment of “quality score” to sscDNA probes. Quality scores of 1–4 for sscDNA were assigned following visual assessment of distribution of polynucleotide size, with representative electropherograms shown here. ( A ) sscDNA synthesised from 50 ng total HUVEC RNA represents an amplification positive control of highest quality achievable. ( B ) sscDNA synthesised from 300 pg total HUVEC RNA amplification positive control of highest quality that could be expected from 300 pg of input RNA. ( C–G ) sscDNA synthesised from 300 pg of total RNA from a vascular endothelial biopsy sample. sscDNA has a electropherogram similar to that seen in the positive controls, peaking at over 500 nts and so was attributed a cDNA quality of 1 (C). sscDNAs progressively shorter in size distribution were designated quality scores of 2, 3 or 4 (D–G). sscDNAs designated lowest quality and therefore not hybridised to GeneChips (C) were of predominantly
    Figure Legend Snippet: Assignment of “quality score” to sscDNA probes. Quality scores of 1–4 for sscDNA were assigned following visual assessment of distribution of polynucleotide size, with representative electropherograms shown here. ( A ) sscDNA synthesised from 50 ng total HUVEC RNA represents an amplification positive control of highest quality achievable. ( B ) sscDNA synthesised from 300 pg total HUVEC RNA amplification positive control of highest quality that could be expected from 300 pg of input RNA. ( C–G ) sscDNA synthesised from 300 pg of total RNA from a vascular endothelial biopsy sample. sscDNA has a electropherogram similar to that seen in the positive controls, peaking at over 500 nts and so was attributed a cDNA quality of 1 (C). sscDNAs progressively shorter in size distribution were designated quality scores of 2, 3 or 4 (D–G). sscDNAs designated lowest quality and therefore not hybridised to GeneChips (C) were of predominantly

    Techniques Used: Amplification, Positive Control

    Bioanalyser electropherograms of cDNA probes synthesised by the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system using a vascular endothelial cell biopsy sample. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and y-axis representing arbitrary signal intensity fluorescence units. Using a vascular endothelial cell biopsy sample a shift to shorter cDNA lengths is apparent compared to 500 pg total HUVEC RNA input in both the ( A ) WT-Ovation FFPE RNA amplification system V2 and the ( B ) WT-Ovation One-Direct RNA amplification system. NTC = no template control.
    Figure Legend Snippet: Bioanalyser electropherograms of cDNA probes synthesised by the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system using a vascular endothelial cell biopsy sample. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and y-axis representing arbitrary signal intensity fluorescence units. Using a vascular endothelial cell biopsy sample a shift to shorter cDNA lengths is apparent compared to 500 pg total HUVEC RNA input in both the ( A ) WT-Ovation FFPE RNA amplification system V2 and the ( B ) WT-Ovation One-Direct RNA amplification system. NTC = no template control.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Amplification, Fluorescence

    Experimental workflow to assess efficiency of NuGen probe generation technologies using low amounts of input RNA. HUVEC total RNA was titrated to cover a range of input RNA from 50 ng–10 pg. 50 ng (n = 1), 500 pg (n = 2) and 250 pg (n = 2) of total RNA was used as input for the WT-Ovation FFPE system V2 while 500 pg (n = 2), 250 pg (n = 2), 100 pg (n = 2), 50 pg (n = 2) and 10 pg (n = 2) were used as input for the WT-Ovation One-Direct system (NuGen Technologies, Inc). All cDNA reactions were purified via Zymo Research Clean and Concentrator™-25 or Qiagen RNeasy MinElute Cleanup kits (WT-Ovation FFPE V2 and WT-Ovation One-Direct systems respectively) as recommended. All purified cDNA probes were assessed for quantity and quality using the Agilent 2100 Bioanalyzer and the Nanodrop-8000 RNA Nano chips. FL-Ovation™ cDNA Biotin Module V2 (NuGEN) was used for fragmentation and biotin labelling of 5 µg of cDNA and used for subsequent hybridisation to Affymetrix HGU133 Plus 2.0 microarrays.
    Figure Legend Snippet: Experimental workflow to assess efficiency of NuGen probe generation technologies using low amounts of input RNA. HUVEC total RNA was titrated to cover a range of input RNA from 50 ng–10 pg. 50 ng (n = 1), 500 pg (n = 2) and 250 pg (n = 2) of total RNA was used as input for the WT-Ovation FFPE system V2 while 500 pg (n = 2), 250 pg (n = 2), 100 pg (n = 2), 50 pg (n = 2) and 10 pg (n = 2) were used as input for the WT-Ovation One-Direct system (NuGen Technologies, Inc). All cDNA reactions were purified via Zymo Research Clean and Concentrator™-25 or Qiagen RNeasy MinElute Cleanup kits (WT-Ovation FFPE V2 and WT-Ovation One-Direct systems respectively) as recommended. All purified cDNA probes were assessed for quantity and quality using the Agilent 2100 Bioanalyzer and the Nanodrop-8000 RNA Nano chips. FL-Ovation™ cDNA Biotin Module V2 (NuGEN) was used for fragmentation and biotin labelling of 5 µg of cDNA and used for subsequent hybridisation to Affymetrix HGU133 Plus 2.0 microarrays.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Purification, Hybridization

    Pearson's Correlation of the signal (MAS5.0) obtained from Affymetrix GeneChips hybridised with cDNA probes synthesised using the NuGen WT-Ovation RNA amplification systems and the effect of reducing RNA input on the resultant Affymetrix GeneChips. ( A ) R 2 values generated when comparing one GeneChip from 50 ng of input HUVEC RNA and duplicate GeneChips from 500 pg and 250 pg of input HUVEC RNA using MAS5.0 analysed data by Pearson's correlation of signal. ( B ) Duplicate GeneChips from 500 pg, 250 pg, 100 pg, 50 pg and 10 pg RNA input compared using MAS5.0 analysed data by Pearson's correlation of signal. C . Principal Component Analysis of all probe sets from GeneChips hybridised with cDNA probes from either WT-Ovation FFPE or WT-Ovation One-Direct RNA amplification systems.
    Figure Legend Snippet: Pearson's Correlation of the signal (MAS5.0) obtained from Affymetrix GeneChips hybridised with cDNA probes synthesised using the NuGen WT-Ovation RNA amplification systems and the effect of reducing RNA input on the resultant Affymetrix GeneChips. ( A ) R 2 values generated when comparing one GeneChip from 50 ng of input HUVEC RNA and duplicate GeneChips from 500 pg and 250 pg of input HUVEC RNA using MAS5.0 analysed data by Pearson's correlation of signal. ( B ) Duplicate GeneChips from 500 pg, 250 pg, 100 pg, 50 pg and 10 pg RNA input compared using MAS5.0 analysed data by Pearson's correlation of signal. C . Principal Component Analysis of all probe sets from GeneChips hybridised with cDNA probes from either WT-Ovation FFPE or WT-Ovation One-Direct RNA amplification systems.

    Techniques Used: Amplification, Generated, Formalin-fixed Paraffin-Embedded

    6) Product Images from "Effects of Secondary Metabolite Extract from Phomopsis occulta on β-Amyloid Aggregation"

    Article Title: Effects of Secondary Metabolite Extract from Phomopsis occulta on β-Amyloid Aggregation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0109438

    Screening of bioactive fractions from Ph. occulta secondary metabolite extracts using an E. coli cell model. A: Inhibitory effect on Aβ42 aggregation of Ph. occulta secondary metabolite extracts. ME: mycelia extracts; BE: broth extracts; 0, 1, 2 and 3 refer to molar salt concentrations in the cultures. B: Inhibitory effect on Aβ42 aggregation of ME0 fractions. ME0-W-F1 to ME0-W-F5 are water soluble fractions separated by column chromatography using Diaion-20 resin and a water/methanol mobile phase. Fraction concentrations were 200 µg/ml in each case; EGCG was used as positive control (100 µg/ml). Values represent mean of means ± SD of four separate experiments, each performed in triplicate.
    Figure Legend Snippet: Screening of bioactive fractions from Ph. occulta secondary metabolite extracts using an E. coli cell model. A: Inhibitory effect on Aβ42 aggregation of Ph. occulta secondary metabolite extracts. ME: mycelia extracts; BE: broth extracts; 0, 1, 2 and 3 refer to molar salt concentrations in the cultures. B: Inhibitory effect on Aβ42 aggregation of ME0 fractions. ME0-W-F1 to ME0-W-F5 are water soluble fractions separated by column chromatography using Diaion-20 resin and a water/methanol mobile phase. Fraction concentrations were 200 µg/ml in each case; EGCG was used as positive control (100 µg/ml). Values represent mean of means ± SD of four separate experiments, each performed in triplicate.

    Techniques Used: Column Chromatography, Positive Control

    ME0-W-F1 reduces aggregation of Aβ42-EGFP and the early onset familial mutation Aβ42E22G-mCherry). (A, B) HEK293 transiently transfected with pcDNA3-Aβ42–EGFP (grey bars) and the Aβ42E22G-mCherry construct pATNRW20 (black bars) and treated with ME0-W-F1 at 17.5 µg/ml (H) and 1.75 µg/ml (L), a positive control (10 µM EGCG) and a negative control (DMSO only). For each construct and treatment, three fields of approximately 200 cells (i.e. n = 200 for each field) were counted for aggregates and odds ratios calculated. Error bars indicate 95% confidence interval for the odds ratio. Treatments of the Aβ42E22G-mCherry transfections with ME0-W-F1 (H and L) and EGCG were all statistically significant with a probability of P
    Figure Legend Snippet: ME0-W-F1 reduces aggregation of Aβ42-EGFP and the early onset familial mutation Aβ42E22G-mCherry). (A, B) HEK293 transiently transfected with pcDNA3-Aβ42–EGFP (grey bars) and the Aβ42E22G-mCherry construct pATNRW20 (black bars) and treated with ME0-W-F1 at 17.5 µg/ml (H) and 1.75 µg/ml (L), a positive control (10 µM EGCG) and a negative control (DMSO only). For each construct and treatment, three fields of approximately 200 cells (i.e. n = 200 for each field) were counted for aggregates and odds ratios calculated. Error bars indicate 95% confidence interval for the odds ratio. Treatments of the Aβ42E22G-mCherry transfections with ME0-W-F1 (H and L) and EGCG were all statistically significant with a probability of P

    Techniques Used: Mutagenesis, Transfection, Construct, Positive Control, Negative Control

    Preparation and analysis of secondary metabolites produced by Phomopsis occulta . A: Proposed strategy for preparation and screening of bioactive fractions from secondary metabolite extracts. B C: Identification of ME0-W-F1 components by TLC analysis. B: visualized by iodide. C: visualized by ninhydrin.
    Figure Legend Snippet: Preparation and analysis of secondary metabolites produced by Phomopsis occulta . A: Proposed strategy for preparation and screening of bioactive fractions from secondary metabolite extracts. B C: Identification of ME0-W-F1 components by TLC analysis. B: visualized by iodide. C: visualized by ninhydrin.

    Techniques Used: Produced, Thin Layer Chromatography

    Effect of ME0-W-F1 on aggregation of Aβ42 analysed by SDS-PAGE and quantitative analysis with Quantitative One. Low (Aβ42: ME0-W-F1 = 1∶1) and high (Aβ42: ME0-W-F1 = 1∶4) concentrations of ME0-W-F1 were used. Values represent mean ± SD of three replicates. The data were evaluated by t -test, * p
    Figure Legend Snippet: Effect of ME0-W-F1 on aggregation of Aβ42 analysed by SDS-PAGE and quantitative analysis with Quantitative One. Low (Aβ42: ME0-W-F1 = 1∶1) and high (Aβ42: ME0-W-F1 = 1∶4) concentrations of ME0-W-F1 were used. Values represent mean ± SD of three replicates. The data were evaluated by t -test, * p

    Techniques Used: SDS Page

    Inhibitory effects of ME0-W-F1 on the transformation of secondary structure of Aβ42 by FT-IR. A: Aβ42 alone; B: Aβ42 with ME0-W-F1; C: change in β-sheet content during incubation with (+) or without (−) ME0-W-F1. Time: 0, 0.5, 1, 2 and 4 days. Values represent mean ± SD of three separate experiments.
    Figure Legend Snippet: Inhibitory effects of ME0-W-F1 on the transformation of secondary structure of Aβ42 by FT-IR. A: Aβ42 alone; B: Aβ42 with ME0-W-F1; C: change in β-sheet content during incubation with (+) or without (−) ME0-W-F1. Time: 0, 0.5, 1, 2 and 4 days. Values represent mean ± SD of three separate experiments.

    Techniques Used: Transformation Assay, Incubation

    Protective effect of ME0-W-F1 in SH-SY5Y cells against cytotoxicity induced by aggregation of Aβ42 (10 µM), as shown by MTT analysis. Four concentrations (µg/ml) were used, with EGCG as positive control. Values represent mean of means ± SD of four separate experiments, each performed in triplicate (i.e. n = 12). The data were evaluated by two-way analysis of variance (ANOVA) followed by a post hoc test. *, **, ***, statistically significant from each other, p
    Figure Legend Snippet: Protective effect of ME0-W-F1 in SH-SY5Y cells against cytotoxicity induced by aggregation of Aβ42 (10 µM), as shown by MTT analysis. Four concentrations (µg/ml) were used, with EGCG as positive control. Values represent mean of means ± SD of four separate experiments, each performed in triplicate (i.e. n = 12). The data were evaluated by two-way analysis of variance (ANOVA) followed by a post hoc test. *, **, ***, statistically significant from each other, p

    Techniques Used: MTT Assay, Positive Control

    7) Product Images from "Analysis of MADS-Box Gene Family Reveals Conservation in Floral Organ ABCDE Model of Moso Bamboo (Phyllostachys edulis)"

    Article Title: Analysis of MADS-Box Gene Family Reveals Conservation in Floral Organ ABCDE Model of Moso Bamboo (Phyllostachys edulis)

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2017.00656

    Analysis of an early flowering phenotype by overexpression of PheMADS15 in Arabidopsis . (A) The flowering phenotype of wild-type (WT), OEPheMADS15-L1 and OEPheMADS15-L2 grown for 4 weeks at 23°C under long-day conditions. (B) Flowering time scored as the number of rosette leaves at flowering of wild-type and transgenic plants at 23°C under long-day conditions. (C) Transcript levels of SOC1 , LFY , and TFL1 in wild-type and transgenic plants (L1 and L2) were evaluated by qPCR. Arabidopsis β-tubulin expression was used as a control. Total RNA from 5-week-old whole- Arabidopsis tissues, including leaves and shoot apex, were used for PheMADS15 , SOC1 , LFY , and TFL1 examination. Error bars indicate standard deviations. Asterisks indicate a statistically significant difference between wild-type and transgenic plants ( P
    Figure Legend Snippet: Analysis of an early flowering phenotype by overexpression of PheMADS15 in Arabidopsis . (A) The flowering phenotype of wild-type (WT), OEPheMADS15-L1 and OEPheMADS15-L2 grown for 4 weeks at 23°C under long-day conditions. (B) Flowering time scored as the number of rosette leaves at flowering of wild-type and transgenic plants at 23°C under long-day conditions. (C) Transcript levels of SOC1 , LFY , and TFL1 in wild-type and transgenic plants (L1 and L2) were evaluated by qPCR. Arabidopsis β-tubulin expression was used as a control. Total RNA from 5-week-old whole- Arabidopsis tissues, including leaves and shoot apex, were used for PheMADS15 , SOC1 , LFY , and TFL1 examination. Error bars indicate standard deviations. Asterisks indicate a statistically significant difference between wild-type and transgenic plants ( P

    Techniques Used: Over Expression, Transgenic Assay, Real-time Polymerase Chain Reaction, Expressing

    8) Product Images from "A new metabolic gene signature in prostate cancer regulated by JMJD3 and EZH2"

    Article Title: A new metabolic gene signature in prostate cancer regulated by JMJD3 and EZH2

    Journal: Oncotarget

    doi: 10.18632/oncotarget.25182

    Effects of treatment and cell line on the gene set expression ANOVA analysis of gene mRNA expression was performed on cell lines (PC-3 (black bars), DU 145 (gray bars) and LNCaP (white bars)) treated with GSK-J4 (neutral bar) or DZNeP (striped bar). Values shown are the average (mean ± S.E.M) from quadruplicate samples for each incubation condition and normalized to control without treatment; y -axis corresponds to relative mRNA quantification in Log. Test designated the statistically significant variables by the letters a, b and c. ( A ) Gene set expression was different according to cell lines, but ( B ) not significant according to treatments. ( C ) Combined effect of cell lines and treatments on whole gene expression. ( D ) Combined effect of cell lines and treatments gene per gene.
    Figure Legend Snippet: Effects of treatment and cell line on the gene set expression ANOVA analysis of gene mRNA expression was performed on cell lines (PC-3 (black bars), DU 145 (gray bars) and LNCaP (white bars)) treated with GSK-J4 (neutral bar) or DZNeP (striped bar). Values shown are the average (mean ± S.E.M) from quadruplicate samples for each incubation condition and normalized to control without treatment; y -axis corresponds to relative mRNA quantification in Log. Test designated the statistically significant variables by the letters a, b and c. ( A ) Gene set expression was different according to cell lines, but ( B ) not significant according to treatments. ( C ) Combined effect of cell lines and treatments on whole gene expression. ( D ) Combined effect of cell lines and treatments gene per gene.

    Techniques Used: Expressing, Incubation

    Effects of GSK-J4 or DZNeP on prostate cancer cell lines ( A ) Cell line viability treated with GSK-J4. PC-3, LNCaP and DU 145 cells were treated for 24 h, 48 h and 72 h with increasing concentrations of GSK-J4. Cell viability was assessed using the XTT assay. The percentage of viable cells was determined as percent of viability of untreated cells. Values shown are the average (mean ± S.E.M) from quadruplicate samples for each incubation condition. ( B – C ) Western-blot analysis of GSK-J4 and DZNeP efficacity. Cells were treated at IC 50 concentration (PC-3: 3.53 μM, LNCaP: 3.93 μM and DU 145: 22.87 μM for 48 h for GSK-J4 and 10 μM for 72 h for DZNeP) (TT) or untreated (N). Quantification representation were expressed as relative fold change in protein expression of JMJD3 and EZH2 in response to GSK-J4 or DZNeP exposure respectively after normalization to GAPDH density.
    Figure Legend Snippet: Effects of GSK-J4 or DZNeP on prostate cancer cell lines ( A ) Cell line viability treated with GSK-J4. PC-3, LNCaP and DU 145 cells were treated for 24 h, 48 h and 72 h with increasing concentrations of GSK-J4. Cell viability was assessed using the XTT assay. The percentage of viable cells was determined as percent of viability of untreated cells. Values shown are the average (mean ± S.E.M) from quadruplicate samples for each incubation condition. ( B – C ) Western-blot analysis of GSK-J4 and DZNeP efficacity. Cells were treated at IC 50 concentration (PC-3: 3.53 μM, LNCaP: 3.93 μM and DU 145: 22.87 μM for 48 h for GSK-J4 and 10 μM for 72 h for DZNeP) (TT) or untreated (N). Quantification representation were expressed as relative fold change in protein expression of JMJD3 and EZH2 in response to GSK-J4 or DZNeP exposure respectively after normalization to GAPDH density.

    Techniques Used: XTT Assay, Incubation, Western Blot, Concentration Assay, Expressing

    Interaction of GSK-J4 and DZNeP on PTEN and AR pathways Effects of JMJD3 and EZH2 inhibitors on key pathways involved in prostate cancer. JMJD3 inhibitor GSK-J4 enhanced H3K27me3 which inhibited PTEN expression and activated AKT; by contrast, DZNeP counteracted these effects. GSK-J4 acts on AR-driven transcription and interferes with proliferation. Arrows indicate an activation, blocked arrows indicate an inhibition and dotted arrows a presumed interaction.
    Figure Legend Snippet: Interaction of GSK-J4 and DZNeP on PTEN and AR pathways Effects of JMJD3 and EZH2 inhibitors on key pathways involved in prostate cancer. JMJD3 inhibitor GSK-J4 enhanced H3K27me3 which inhibited PTEN expression and activated AKT; by contrast, DZNeP counteracted these effects. GSK-J4 acts on AR-driven transcription and interferes with proliferation. Arrows indicate an activation, blocked arrows indicate an inhibition and dotted arrows a presumed interaction.

    Techniques Used: Expressing, Activation Assay, Inhibition

    9) Product Images from "Independent Action between DvSnf7 RNA and Cry3Bb1 Protein in Southern Corn Rootworm, Diabrotica undecimpunctata howardi and Colorado Potato Beetle, Leptinotarsa decemlineata"

    Article Title: Independent Action between DvSnf7 RNA and Cry3Bb1 Protein in Southern Corn Rootworm, Diabrotica undecimpunctata howardi and Colorado Potato Beetle, Leptinotarsa decemlineata

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0118622

    Concentration response curves for growth inhibition activity of DvSnf7 (A) and Cry3Bb1 (B) and for mortality of DvSnf7 and Cry3Bb1 (C) to SCR in 12-day diet incorporation bioassays. Data points represent mean values with standard errors of six replicated bioassays for growth inhibition curves (A and B) and of five replicated bioassays for mortality curves (C). The concentration-mortality response curves were plotted with log transformation of concentrations (X-abscissa) plotted against the percent mortality (Y-abscissa).
    Figure Legend Snippet: Concentration response curves for growth inhibition activity of DvSnf7 (A) and Cry3Bb1 (B) and for mortality of DvSnf7 and Cry3Bb1 (C) to SCR in 12-day diet incorporation bioassays. Data points represent mean values with standard errors of six replicated bioassays for growth inhibition curves (A and B) and of five replicated bioassays for mortality curves (C). The concentration-mortality response curves were plotted with log transformation of concentrations (X-abscissa) plotted against the percent mortality (Y-abscissa).

    Techniques Used: Concentration Assay, Inhibition, Activity Assay, Transformation Assay

    Observed growth inhibition (GI) with standard errors (SEs) of Cry3Bb1 and DvSnf7 and their mixtures at levels of low response (A), medium response (B) and high response (C) in 12-days diet incorporation bioassays. The predicted GI of the mixture in each level was calculated based on the GIs of two individuals using the RA model. The average GI and associated SEs were calculated from three replicates.
    Figure Legend Snippet: Observed growth inhibition (GI) with standard errors (SEs) of Cry3Bb1 and DvSnf7 and their mixtures at levels of low response (A), medium response (B) and high response (C) in 12-days diet incorporation bioassays. The predicted GI of the mixture in each level was calculated based on the GIs of two individuals using the RA model. The average GI and associated SEs were calculated from three replicates.

    Techniques Used: Inhibition

    Concentration response curves of CPB mortality for Cry3Bb1 alone (Δ) and Cry3Bb1 in the presence of DvSnf7 RNA (○) at a fixed concentration of 1000 ng/ml diet in 12-day diet bioassays. The responses were plotted based on the tested Cry3Bb1 concentrations, rather than the mixture concentrations. Data points represent mean mortality with standard errors of three replicated bioassays. LC 50 values and associated 95% confidence intervals are listed in Table 2 .
    Figure Legend Snippet: Concentration response curves of CPB mortality for Cry3Bb1 alone (Δ) and Cry3Bb1 in the presence of DvSnf7 RNA (○) at a fixed concentration of 1000 ng/ml diet in 12-day diet bioassays. The responses were plotted based on the tested Cry3Bb1 concentrations, rather than the mixture concentrations. Data points represent mean mortality with standard errors of three replicated bioassays. LC 50 values and associated 95% confidence intervals are listed in Table 2 .

    Techniques Used: Concentration Assay

    Observed concentration-mortality responses of Cry3Bb1 and DvSnf7 to the SCR. (A) Cry3Bb1 when applied alone (■) as compared to its mixture with a fixed sub-lethal concentration of DvSnf7 (●) and (B) DvSnf7 when applied alone (▲) as compared to its mixture with a fixed sub-lethal concentration of Cry3Bb1 (▲). The responses were plotted based on the tested Cry3Bb1 (A) or DvSnf7 concentrations (B), rather than the mixture concentrations. Data points represent mean mortality with standard errors of three replicated bioassays. LC 50 values and associated 95% confidence intervals are listed in Table 1 .
    Figure Legend Snippet: Observed concentration-mortality responses of Cry3Bb1 and DvSnf7 to the SCR. (A) Cry3Bb1 when applied alone (■) as compared to its mixture with a fixed sub-lethal concentration of DvSnf7 (●) and (B) DvSnf7 when applied alone (▲) as compared to its mixture with a fixed sub-lethal concentration of Cry3Bb1 (▲). The responses were plotted based on the tested Cry3Bb1 (A) or DvSnf7 concentrations (B), rather than the mixture concentrations. Data points represent mean mortality with standard errors of three replicated bioassays. LC 50 values and associated 95% confidence intervals are listed in Table 1 .

    Techniques Used: Concentration Assay

    Time-to-effect relationships for SCR growth inhibition (A) and mortality (B) of DvSnf7 (●) and Cry3Bb1 (■) in 12-day diet incorporation bioassays. Arrows indicate the onset of significant growth inhibition or mortality. The time-to-effect for growth inhibition was assessed at a concentration of 0.012 μg DvSnf7/ml diet and 25 μg Cry3Bb1/ml diet which approximated the GI 80 concentration. Each treatment included 15 plates and each plate had 42 individually housed SCR larvae. Water and buffer controls were included with the same number of plates and same number of SCR for each plate. Growth inhibition was calculated based on the reduction in mean body mass of surviving SCR in relation to the appropriate control. The time-to-effect for mortality was assessed at concentrations of 250 μg Cry3Bb1/ml diet and 0.050 μg DvSnf7/ml diet which approximated the LC 95 concentration. Mortality was recorded daily from the same plate with 36 individually housed SCR larvae and was corrected for background mortality.
    Figure Legend Snippet: Time-to-effect relationships for SCR growth inhibition (A) and mortality (B) of DvSnf7 (●) and Cry3Bb1 (■) in 12-day diet incorporation bioassays. Arrows indicate the onset of significant growth inhibition or mortality. The time-to-effect for growth inhibition was assessed at a concentration of 0.012 μg DvSnf7/ml diet and 25 μg Cry3Bb1/ml diet which approximated the GI 80 concentration. Each treatment included 15 plates and each plate had 42 individually housed SCR larvae. Water and buffer controls were included with the same number of plates and same number of SCR for each plate. Growth inhibition was calculated based on the reduction in mean body mass of surviving SCR in relation to the appropriate control. The time-to-effect for mortality was assessed at concentrations of 250 μg Cry3Bb1/ml diet and 0.050 μg DvSnf7/ml diet which approximated the LC 95 concentration. Mortality was recorded daily from the same plate with 36 individually housed SCR larvae and was corrected for background mortality.

    Techniques Used: Inhibition, Concentration Assay

    10) Product Images from "Mitochondrial dysfunction induced by leflunomide and its active metabolite"

    Article Title: Mitochondrial dysfunction induced by leflunomide and its active metabolite

    Journal: Toxicology

    doi: 10.1016/j.tox.2018.02.003

    Mitochondrial dysfunction modulates the level of ER stress. (A) HepG2 cells were pre-treated with MPTP inhibitor bongkrekic acid (BA) (10 μM) for 2 h before exposure to 200 μM of leflunomide for 6 h. Western blot showed that the presence of BA attenuated the leflunomide-induced activation of ATF4 and CHOP. (B) HepG2 cells were pre-treated with MPTP inhibitor cyclosporine A (CsA) (1 μM) for 2 h before exposure to 200 μM of leflunomide for 6 h. Representative Western blot showed that CsA failed to alleviate the ER stress induced by leflunomide. (C) Glucose-grown and galactose-grown HepG2 cells were exposed to 50 to 300 μM of leflunomide for 6 h. Western blot showed that galactose-grown HepG2 cells exhibited more prominent increase in both ATF4 and CHOP protein levels. Similar Western results were obtained from three independent experiments. Intensities of bands were normalized to the amount of GAPDH. *p
    Figure Legend Snippet: Mitochondrial dysfunction modulates the level of ER stress. (A) HepG2 cells were pre-treated with MPTP inhibitor bongkrekic acid (BA) (10 μM) for 2 h before exposure to 200 μM of leflunomide for 6 h. Western blot showed that the presence of BA attenuated the leflunomide-induced activation of ATF4 and CHOP. (B) HepG2 cells were pre-treated with MPTP inhibitor cyclosporine A (CsA) (1 μM) for 2 h before exposure to 200 μM of leflunomide for 6 h. Representative Western blot showed that CsA failed to alleviate the ER stress induced by leflunomide. (C) Glucose-grown and galactose-grown HepG2 cells were exposed to 50 to 300 μM of leflunomide for 6 h. Western blot showed that galactose-grown HepG2 cells exhibited more prominent increase in both ATF4 and CHOP protein levels. Similar Western results were obtained from three independent experiments. Intensities of bands were normalized to the amount of GAPDH. *p

    Techniques Used: Western Blot, Activation Assay

    Cytotoxicity of leflunomide (A) and A77 1726 (B) in human hepatoma HepG2 cells. HepG2 cells were treated with DMSO as vehicle control and leflunomide or A77 1726 for 2, 6, and 24 h. Cytotoxicity was assessed by cellular ATP depletion (bars) and LDH release (lines). Data were analyzed for statistical significance using one-way ANOVA followed by Dunnett’s test. Data are expressed as mean ± SD (n = 3). Statistical significance compared with control: **, p
    Figure Legend Snippet: Cytotoxicity of leflunomide (A) and A77 1726 (B) in human hepatoma HepG2 cells. HepG2 cells were treated with DMSO as vehicle control and leflunomide or A77 1726 for 2, 6, and 24 h. Cytotoxicity was assessed by cellular ATP depletion (bars) and LDH release (lines). Data were analyzed for statistical significance using one-way ANOVA followed by Dunnett’s test. Data are expressed as mean ± SD (n = 3). Statistical significance compared with control: **, p

    Techniques Used:

    Effects of leflunomide (A) and A77 1726 (B) on mitochondrial function assessed by glucose/galactose assay. Dose responses at 24 h for glucose-grown (25 mM) (open symbol) and galactose-grown (10 mM) (filled symbol) HepG2 cells treated with leflunomide (A) and A77 1726 (B). Data were analyzed for statistical significance using two-tailed unpaired Student’s t -test. Data are expressed as mean ± SD (n = 3). Statistical significance compared between two culture conditions: *** p
    Figure Legend Snippet: Effects of leflunomide (A) and A77 1726 (B) on mitochondrial function assessed by glucose/galactose assay. Dose responses at 24 h for glucose-grown (25 mM) (open symbol) and galactose-grown (10 mM) (filled symbol) HepG2 cells treated with leflunomide (A) and A77 1726 (B). Data were analyzed for statistical significance using two-tailed unpaired Student’s t -test. Data are expressed as mean ± SD (n = 3). Statistical significance compared between two culture conditions: *** p

    Techniques Used: Galactose Assay, Two Tailed Test

    Effects of mitochondrial permeability transition (MPT) inhibitors on ATP depletion, LDH release and mitochondrial membrane potential (MMP) induced by leflunomide (A) and A771726 (B). HepG2 cells were pre-treated with 1 μM cyclosporine A (CsA) or 10 μM bongkrekic acid (BA) followed by a 24-h exposure to leflunomide or A77 1726. MMP was assessed by the JC-1 fluorescent probe. Data were analyzed for statistical significance using one-way ANOVA followed by Dunnett’s test. Data are expressed as mean ± SD (n = 3). Statistical significance compared with control: **, p
    Figure Legend Snippet: Effects of mitochondrial permeability transition (MPT) inhibitors on ATP depletion, LDH release and mitochondrial membrane potential (MMP) induced by leflunomide (A) and A771726 (B). HepG2 cells were pre-treated with 1 μM cyclosporine A (CsA) or 10 μM bongkrekic acid (BA) followed by a 24-h exposure to leflunomide or A77 1726. MMP was assessed by the JC-1 fluorescent probe. Data were analyzed for statistical significance using one-way ANOVA followed by Dunnett’s test. Data are expressed as mean ± SD (n = 3). Statistical significance compared with control: **, p

    Techniques Used: Permeability

    11) Product Images from "Genome-wide and single-cell analyses reveal a context dependent relationship between CBP recruitment and gene expression"

    Article Title: Genome-wide and single-cell analyses reveal a context dependent relationship between CBP recruitment and gene expression

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gku827

    Differential HIF1α, CBP or p300 recruitment does not explain variable dependence on CBP and p300 for DP-inducible expression. ( A – D ) qRT-PCR on WT ( n = 2) and CBP Δflox/Δflox ;p300 Δflox/Δflox (dKO, n = 3) MEFs treated for 3 h with DP or ethanol vehicle (EtOH, mean ± SEM). Expression between samples normalized to expression of Ubc . ( E – H ) ChIP using antibodies to proteins indicated (NRS is normal rabbit serum control) at hypoxia response elements located as indicated relative to gene TSS following 2 h treatment with DP. ChIP signals are normalized to the input signal for each sample. ( n = 2, mean ± SEM).
    Figure Legend Snippet: Differential HIF1α, CBP or p300 recruitment does not explain variable dependence on CBP and p300 for DP-inducible expression. ( A – D ) qRT-PCR on WT ( n = 2) and CBP Δflox/Δflox ;p300 Δflox/Δflox (dKO, n = 3) MEFs treated for 3 h with DP or ethanol vehicle (EtOH, mean ± SEM). Expression between samples normalized to expression of Ubc . ( E – H ) ChIP using antibodies to proteins indicated (NRS is normal rabbit serum control) at hypoxia response elements located as indicated relative to gene TSS following 2 h treatment with DP. ChIP signals are normalized to the input signal for each sample. ( n = 2, mean ± SEM).

    Techniques Used: Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation

    CBP recruitment does not guarantee induction of HIF target genes. ( A ) IGV browser views of CBP ChIP-seq data showing DP-inducible recruitment of CBP to the promoter of Ddah1 in mouse embryonic fibroblasts (MEFs). CBP Δflox/Δflox MEFs provide a negative control for the CBP antibody used. Read height for each track set to 150. ( B ) ChIP with pooled antibodies against CBP and p300 or normal rabbit serum (NRS) control at Ddah1 promoter in WT and CBP Δflox/Δflo x ;p300 Δflox/Δflox (dKO) MEFs. ( C ) ChIP of HIF1α and Pol II at Ddah1 promoter in WT MEFs. ( D ) qRT-PCR of Ddah1 message in WT MEFs treated with DP for 3, 8 and 24 h. No statistical differences were found by one-way ANOVA with Tukey pairwise post tests. ( E ) qRT-PCR primary transcript analysis of Ddah1 unspliced primary transcript in WT MEFs. DNaseI-treated samples were prepared with and without reverse transcription (RT) as a control for potential genomic DNA contamination. No statistical differences were found by one-way ANOVA with Tukey pairwise post tests of reverse transcribed samples.
    Figure Legend Snippet: CBP recruitment does not guarantee induction of HIF target genes. ( A ) IGV browser views of CBP ChIP-seq data showing DP-inducible recruitment of CBP to the promoter of Ddah1 in mouse embryonic fibroblasts (MEFs). CBP Δflox/Δflox MEFs provide a negative control for the CBP antibody used. Read height for each track set to 150. ( B ) ChIP with pooled antibodies against CBP and p300 or normal rabbit serum (NRS) control at Ddah1 promoter in WT and CBP Δflox/Δflo x ;p300 Δflox/Δflox (dKO) MEFs. ( C ) ChIP of HIF1α and Pol II at Ddah1 promoter in WT MEFs. ( D ) qRT-PCR of Ddah1 message in WT MEFs treated with DP for 3, 8 and 24 h. No statistical differences were found by one-way ANOVA with Tukey pairwise post tests. ( E ) qRT-PCR primary transcript analysis of Ddah1 unspliced primary transcript in WT MEFs. DNaseI-treated samples were prepared with and without reverse transcription (RT) as a control for potential genomic DNA contamination. No statistical differences were found by one-way ANOVA with Tukey pairwise post tests of reverse transcribed samples.

    Techniques Used: Chromatin Immunoprecipitation, Negative Control, Quantitative RT-PCR

    12) Product Images from "Mice harboring the human SLC30A8 R138X loss-of-function mutation have increased insulin secretory capacity"

    Article Title: Mice harboring the human SLC30A8 R138X loss-of-function mutation have increased insulin secretory capacity

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1721418115

    Analysis of Slc30a8 RNA and protein in islets from male R138X mice on chow diet. ( A ) Slc30a8 RNA in situ hybridization of pancreatic islets isolated from wild-type, knockout, and R138X mice. KO islets were used as negative control. Red, glucagon RNA; green, insulin RNA; white, Slc30a8 RNA. ( B ) Quantification of islet Slc30a8 RNA levels using qPCR analysis. n.d., not detected. ( C ) Western blot of islets isolated from chow-fed WT, KO, and R138X mice. KO islets were used as negative control. The arrow indicates SLC30A8 protein; asterisks denote unspecific bands. ( D ) Dithizone staining of pancreatic islets isolated from WT, KO, and R138X mice.
    Figure Legend Snippet: Analysis of Slc30a8 RNA and protein in islets from male R138X mice on chow diet. ( A ) Slc30a8 RNA in situ hybridization of pancreatic islets isolated from wild-type, knockout, and R138X mice. KO islets were used as negative control. Red, glucagon RNA; green, insulin RNA; white, Slc30a8 RNA. ( B ) Quantification of islet Slc30a8 RNA levels using qPCR analysis. n.d., not detected. ( C ) Western blot of islets isolated from chow-fed WT, KO, and R138X mice. KO islets were used as negative control. The arrow indicates SLC30A8 protein; asterisks denote unspecific bands. ( D ) Dithizone staining of pancreatic islets isolated from WT, KO, and R138X mice.

    Techniques Used: Mouse Assay, RNA In Situ Hybridization, Isolation, Knock-Out, Negative Control, Real-time Polymerase Chain Reaction, Western Blot, Staining

    13) Product Images from "Role of miR-27a, miR-181a and miR-20b in gastric cancer hypoxia-induced chemoresistance"

    Article Title: Role of miR-27a, miR-181a and miR-20b in gastric cancer hypoxia-induced chemoresistance

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2016.1139244

    Median levels ofmiR-27a, miR-181a and miR-20b in GC patients with PD vs those with DCR (A). Percentage of miR-27a, miR-181a and miR-20b overexpression in GC patients with PD vs those with DCR (B). Median levels of HIF1A, MDR1 and HIPK2 in GC patients
    Figure Legend Snippet: Median levels ofmiR-27a, miR-181a and miR-20b in GC patients with PD vs those with DCR (A). Percentage of miR-27a, miR-181a and miR-20b overexpression in GC patients with PD vs those with DCR (B). Median levels of HIF1A, MDR1 and HIPK2 in GC patients

    Techniques Used: Over Expression

    Correlation analysis between HIF1A and miR-20bexpression (A) MDR1 and miR-27a levels (B) and HIPK2 and miR-181a (C) and HIPK2 and miR-27a (D) in GC patients.
    Figure Legend Snippet: Correlation analysis between HIF1A and miR-20bexpression (A) MDR1 and miR-27a levels (B) and HIPK2 and miR-181a (C) and HIPK2 and miR-27a (D) in GC patients.

    Techniques Used:

    14) Product Images from "Endoplasmic reticulum stress and MAPK signaling pathway activation underlie leflunomide-induced toxicity in HepG2 Cells"

    Article Title: Endoplasmic reticulum stress and MAPK signaling pathway activation underlie leflunomide-induced toxicity in HepG2 Cells

    Journal: Toxicology

    doi: 10.1016/j.tox.2017.10.002

    Leflunomide triggers ER stress and UPR in HepG2 cells ( A , B ) Total cellular proteins from HepG2 cells were extracted after 6 h leflunomide treatment at the concentrations indicated. Protein levels of CHOP, ATF-4, spliced XBP1 (XBP1s), p-eIF2α, total eIF2α, and GAPDH as internal control were assessed by Western blotting ( A ) and quantified ( B ). ( C ) HepG2-Gluc-Fluc cells were exposed to 25–300 µM of leflunomide for 2, 6, and 24 h, and the activity of Gluc was measured. ( D ) HepG2 cells were treated with 4-PBA for 2 h before exposure to leflunomide for additional 24 h. The protective effect of 4-PBA was assessed by cellular ATP content. ( E ) Representative Western blots showed that 4-PBA attenuated leflunomide-induced increase of CHOP and ATF-4. The results shown are mean ± S.D. from three independent experiments. * p
    Figure Legend Snippet: Leflunomide triggers ER stress and UPR in HepG2 cells ( A , B ) Total cellular proteins from HepG2 cells were extracted after 6 h leflunomide treatment at the concentrations indicated. Protein levels of CHOP, ATF-4, spliced XBP1 (XBP1s), p-eIF2α, total eIF2α, and GAPDH as internal control were assessed by Western blotting ( A ) and quantified ( B ). ( C ) HepG2-Gluc-Fluc cells were exposed to 25–300 µM of leflunomide for 2, 6, and 24 h, and the activity of Gluc was measured. ( D ) HepG2 cells were treated with 4-PBA for 2 h before exposure to leflunomide for additional 24 h. The protective effect of 4-PBA was assessed by cellular ATP content. ( E ) Representative Western blots showed that 4-PBA attenuated leflunomide-induced increase of CHOP and ATF-4. The results shown are mean ± S.D. from three independent experiments. * p

    Techniques Used: Western Blot, Activity Assay

    15) Product Images from "Establishment of LIF-Dependent Human iPS Cells Closely Related to Basic FGF-Dependent Authentic iPS Cells"

    Article Title: Establishment of LIF-Dependent Human iPS Cells Closely Related to Basic FGF-Dependent Authentic iPS Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0039022

    Genome-wide gene expression patterns comparing various pluripotent stem cells. (A) Scatter plots comparing levels of whole transcripts obtained from two WA09 ESC cultures, two dermal fibroblast cultures, two lines each for L-iPSCs, F-iPSCs and LF-iPSCs. Two independently cultured cells were used for WA09 cells. All these iPSCs were established from fibroblasts obtained from a single adult female. Two different cultures of fibroblasts obtained from the same individual were used. (B) Cluster analysis of the genome-wide gene expression patterns of cells analyzed in (A). (C) Scatter plots comparing levels of whole transcripts between L-iPSCs and other cell types used in this study. Average values obtained from two cultures or two lines for each cell type were used.
    Figure Legend Snippet: Genome-wide gene expression patterns comparing various pluripotent stem cells. (A) Scatter plots comparing levels of whole transcripts obtained from two WA09 ESC cultures, two dermal fibroblast cultures, two lines each for L-iPSCs, F-iPSCs and LF-iPSCs. Two independently cultured cells were used for WA09 cells. All these iPSCs were established from fibroblasts obtained from a single adult female. Two different cultures of fibroblasts obtained from the same individual were used. (B) Cluster analysis of the genome-wide gene expression patterns of cells analyzed in (A). (C) Scatter plots comparing levels of whole transcripts between L-iPSCs and other cell types used in this study. Average values obtained from two cultures or two lines for each cell type were used.

    Techniques Used: Genome Wide, Expressing, Cell Culture

    Characterization of gene expression patterns, karyotype, and teratoma formation in L-iPSCs prepared with M 3 O-SKM. (A) Immunofluorescence staining of L-iPSCs with antibodies against OCT4, NANOG, TRA1-60 and TRA1-81 on day 120. Alkaline phosphatase staining is also shown. Bar, 100 µm. (B) Quantitative RT-PCR analysis of genes that are typically expressed in pluripotent stem cells. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (C) Karyotype analysis of an L-iPSC on day 120. (D) Haematoxylin and eosin staining of histological sections of teratomas derived from L-iPSCs. Bar, 200 µm.
    Figure Legend Snippet: Characterization of gene expression patterns, karyotype, and teratoma formation in L-iPSCs prepared with M 3 O-SKM. (A) Immunofluorescence staining of L-iPSCs with antibodies against OCT4, NANOG, TRA1-60 and TRA1-81 on day 120. Alkaline phosphatase staining is also shown. Bar, 100 µm. (B) Quantitative RT-PCR analysis of genes that are typically expressed in pluripotent stem cells. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (C) Karyotype analysis of an L-iPSC on day 120. (D) Haematoxylin and eosin staining of histological sections of teratomas derived from L-iPSCs. Bar, 200 µm.

    Techniques Used: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Derivative Assay

    Characterization of colony morphology, gene expression patterns, and sensitivity to an FGF receptor inhibitor in L-iPSCs obtained with M 3 O-SKM. (A) Phase contrast images of L-iPSC colonies obtained on day 90. Bar, 100 µm. (B) Immunofluorescence staining of an L-iPSC colony with antibodies against NANOG and SSEA4 on day 28. Bar, 100 µm. (C) Quantitative RT-PCR analysis of the OCT4 , SOX2 , KLF4 and c- MYC genes in various pluripotent stem cells on day 120. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (D) RT-PCR analysis of the expression of M 3 O in day-3 fibroblasts transduced with M 3 O-SKM, day-120 L-iPSCs, and parent fibroblasts. Expression of GAPDH mRNA was monitored as a control. Bar, 100 µm. (E) Effects of SU5402 on iPSC colony formation with LIF and bFGF. SU5402 dissolved in dimethyl sulfoxide (DMSO) was added to culture medium between day 8 and 10, and the colonies were counted on day 12. DMSO was used as a control.
    Figure Legend Snippet: Characterization of colony morphology, gene expression patterns, and sensitivity to an FGF receptor inhibitor in L-iPSCs obtained with M 3 O-SKM. (A) Phase contrast images of L-iPSC colonies obtained on day 90. Bar, 100 µm. (B) Immunofluorescence staining of an L-iPSC colony with antibodies against NANOG and SSEA4 on day 28. Bar, 100 µm. (C) Quantitative RT-PCR analysis of the OCT4 , SOX2 , KLF4 and c- MYC genes in various pluripotent stem cells on day 120. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (D) RT-PCR analysis of the expression of M 3 O in day-3 fibroblasts transduced with M 3 O-SKM, day-120 L-iPSCs, and parent fibroblasts. Expression of GAPDH mRNA was monitored as a control. Bar, 100 µm. (E) Effects of SU5402 on iPSC colony formation with LIF and bFGF. SU5402 dissolved in dimethyl sulfoxide (DMSO) was added to culture medium between day 8 and 10, and the colonies were counted on day 12. DMSO was used as a control.

    Techniques Used: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Transduction

    Characterization of L-iPSCs by using gene expression analyses and immunofluorescence staining of H3K27me3. (A) Quantitative RT-PCR analysis of genes that are typically expressed in mouse naïve ( REX1 and STELLA ) or primed ( FGF5 and T ) pluripotent stem cells. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (B) Immunofluorescence staining of H3K27me3 as an indicator for X chromosome inactivation. Parent fibroblasts were stained as a positive control. Nuclei surrounded by squares were magnified (left panels). Arrows indicate H3K27me-positive areas, corresponding to inactive X chromosomes. Bar, 50 µm.
    Figure Legend Snippet: Characterization of L-iPSCs by using gene expression analyses and immunofluorescence staining of H3K27me3. (A) Quantitative RT-PCR analysis of genes that are typically expressed in mouse naïve ( REX1 and STELLA ) or primed ( FGF5 and T ) pluripotent stem cells. Expression levels of each gene were normalized to GAPDH. The normalized value for ESCs was defined as 1.0. Mean + SD obtained from three independent experiments are shown. (B) Immunofluorescence staining of H3K27me3 as an indicator for X chromosome inactivation. Parent fibroblasts were stained as a positive control. Nuclei surrounded by squares were magnified (left panels). Arrows indicate H3K27me-positive areas, corresponding to inactive X chromosomes. Bar, 50 µm.

    Techniques Used: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Positive Control

    16) Product Images from "Global Array-Based Transcriptomics from Minimal Input RNA Utilising an Optimal RNA Isolation Process Combined with SPIA cDNA Probes"

    Article Title: Global Array-Based Transcriptomics from Minimal Input RNA Utilising an Optimal RNA Isolation Process Combined with SPIA cDNA Probes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0017625

    Assignment of “quality score” to sscDNA probes. Quality scores of 1–4 for sscDNA were assigned following visual assessment of distribution of polynucleotide size, with representative electropherograms shown here. ( A ) sscDNA synthesised from 50 ng total HUVEC RNA represents an amplification positive control of highest quality achievable. ( B ) sscDNA synthesised from 300 pg total HUVEC RNA amplification positive control of highest quality that could be expected from 300 pg of input RNA. ( C–G ) sscDNA synthesised from 300 pg of total RNA from a vascular endothelial biopsy sample. sscDNA has a electropherogram similar to that seen in the positive controls, peaking at over 500 nts and so was attributed a cDNA quality of 1 (C). sscDNAs progressively shorter in size distribution were designated quality scores of 2, 3 or 4 (D–G). sscDNAs designated lowest quality and therefore not hybridised to GeneChips (C) were of predominantly
    Figure Legend Snippet: Assignment of “quality score” to sscDNA probes. Quality scores of 1–4 for sscDNA were assigned following visual assessment of distribution of polynucleotide size, with representative electropherograms shown here. ( A ) sscDNA synthesised from 50 ng total HUVEC RNA represents an amplification positive control of highest quality achievable. ( B ) sscDNA synthesised from 300 pg total HUVEC RNA amplification positive control of highest quality that could be expected from 300 pg of input RNA. ( C–G ) sscDNA synthesised from 300 pg of total RNA from a vascular endothelial biopsy sample. sscDNA has a electropherogram similar to that seen in the positive controls, peaking at over 500 nts and so was attributed a cDNA quality of 1 (C). sscDNAs progressively shorter in size distribution were designated quality scores of 2, 3 or 4 (D–G). sscDNAs designated lowest quality and therefore not hybridised to GeneChips (C) were of predominantly

    Techniques Used: Amplification, Positive Control

    Bioanalyser electropherograms of cDNA probes synthesised by the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system using a vascular endothelial cell biopsy sample. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and y-axis representing arbitrary signal intensity fluorescence units. Using a vascular endothelial cell biopsy sample a shift to shorter cDNA lengths is apparent compared to 500 pg total HUVEC RNA input in both the ( A ) WT-Ovation FFPE RNA amplification system V2 and the ( B ) WT-Ovation One-Direct RNA amplification system. NTC = no template control.
    Figure Legend Snippet: Bioanalyser electropherograms of cDNA probes synthesised by the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system using a vascular endothelial cell biopsy sample. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and y-axis representing arbitrary signal intensity fluorescence units. Using a vascular endothelial cell biopsy sample a shift to shorter cDNA lengths is apparent compared to 500 pg total HUVEC RNA input in both the ( A ) WT-Ovation FFPE RNA amplification system V2 and the ( B ) WT-Ovation One-Direct RNA amplification system. NTC = no template control.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Amplification, Fluorescence

    Bioanalyser electropherograms of cDNA probes generated using the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and the y-axis representing arbitrary signal intensity fluorescence units. Electropherograms of representative cDNA from each WT-Ovation system are shown. ( A ) WT-Ovation FFPE system sscDNA synthesised from 50 ng of RNA is distributed 500–1000 nucleotides. The sscDNA synthesised from 500 pg and 250 pg of RNA input in the WT-Ovation FFPE system show a similar distribution to the sscDNA synthesised from 50 ng of RNA. ( B ) The majority of dscDNA fragments synthesised in the WT-Ovation One-Direct system average in length at approximately 100–150 nucleotides. RNA input level does not influence polynucleotide length (500 and 250 pg input shown). A significant difference in polynucleotide distribution is observed in 500 pg and 250 pg input RNA reactions depending on which WT-Ovation system used for probe synthesis. ( C ) In the WT-Ovation One-Direct system, the dscDNA probes generated from a reaction containing RNA template is distinct from that generated in a parallel no template control.
    Figure Legend Snippet: Bioanalyser electropherograms of cDNA probes generated using the WT-Ovation FFPE RNA amplification system V2 and the WT-Ovation One-Direct RNA amplification system. cDNA quality assessed by distribution of size with the x-axis representing polynucleotide length and the y-axis representing arbitrary signal intensity fluorescence units. Electropherograms of representative cDNA from each WT-Ovation system are shown. ( A ) WT-Ovation FFPE system sscDNA synthesised from 50 ng of RNA is distributed 500–1000 nucleotides. The sscDNA synthesised from 500 pg and 250 pg of RNA input in the WT-Ovation FFPE system show a similar distribution to the sscDNA synthesised from 50 ng of RNA. ( B ) The majority of dscDNA fragments synthesised in the WT-Ovation One-Direct system average in length at approximately 100–150 nucleotides. RNA input level does not influence polynucleotide length (500 and 250 pg input shown). A significant difference in polynucleotide distribution is observed in 500 pg and 250 pg input RNA reactions depending on which WT-Ovation system used for probe synthesis. ( C ) In the WT-Ovation One-Direct system, the dscDNA probes generated from a reaction containing RNA template is distinct from that generated in a parallel no template control.

    Techniques Used: Generated, Formalin-fixed Paraffin-Embedded, Amplification, Fluorescence

    Experimental workflow to assess efficiency of NuGen probe generation technologies using low amounts of input RNA. HUVEC total RNA was titrated to cover a range of input RNA from 50 ng–10 pg. 50 ng (n = 1), 500 pg (n = 2) and 250 pg (n = 2) of total RNA was used as input for the WT-Ovation FFPE system V2 while 500 pg (n = 2), 250 pg (n = 2), 100 pg (n = 2), 50 pg (n = 2) and 10 pg (n = 2) were used as input for the WT-Ovation One-Direct system (NuGen Technologies, Inc). All cDNA reactions were purified via Zymo Research Clean and Concentrator™-25 or Qiagen RNeasy MinElute Cleanup kits (WT-Ovation FFPE V2 and WT-Ovation One-Direct systems respectively) as recommended. All purified cDNA probes were assessed for quantity and quality using the Agilent 2100 Bioanalyzer and the Nanodrop-8000 RNA Nano chips. FL-Ovation™ cDNA Biotin Module V2 (NuGEN) was used for fragmentation and biotin labelling of 5 µg of cDNA and used for subsequent hybridisation to Affymetrix HGU133 Plus 2.0 microarrays.
    Figure Legend Snippet: Experimental workflow to assess efficiency of NuGen probe generation technologies using low amounts of input RNA. HUVEC total RNA was titrated to cover a range of input RNA from 50 ng–10 pg. 50 ng (n = 1), 500 pg (n = 2) and 250 pg (n = 2) of total RNA was used as input for the WT-Ovation FFPE system V2 while 500 pg (n = 2), 250 pg (n = 2), 100 pg (n = 2), 50 pg (n = 2) and 10 pg (n = 2) were used as input for the WT-Ovation One-Direct system (NuGen Technologies, Inc). All cDNA reactions were purified via Zymo Research Clean and Concentrator™-25 or Qiagen RNeasy MinElute Cleanup kits (WT-Ovation FFPE V2 and WT-Ovation One-Direct systems respectively) as recommended. All purified cDNA probes were assessed for quantity and quality using the Agilent 2100 Bioanalyzer and the Nanodrop-8000 RNA Nano chips. FL-Ovation™ cDNA Biotin Module V2 (NuGEN) was used for fragmentation and biotin labelling of 5 µg of cDNA and used for subsequent hybridisation to Affymetrix HGU133 Plus 2.0 microarrays.

    Techniques Used: Formalin-fixed Paraffin-Embedded, Purification, Hybridization

    Pearson's Correlation of the signal (MAS5.0) obtained from Affymetrix GeneChips hybridised with cDNA probes synthesised using the NuGen WT-Ovation RNA amplification systems and the effect of reducing RNA input on the resultant Affymetrix GeneChips. ( A ) R 2 values generated when comparing one GeneChip from 50 ng of input HUVEC RNA and duplicate GeneChips from 500 pg and 250 pg of input HUVEC RNA using MAS5.0 analysed data by Pearson's correlation of signal. ( B ) Duplicate GeneChips from 500 pg, 250 pg, 100 pg, 50 pg and 10 pg RNA input compared using MAS5.0 analysed data by Pearson's correlation of signal. C . Principal Component Analysis of all probe sets from GeneChips hybridised with cDNA probes from either WT-Ovation FFPE or WT-Ovation One-Direct RNA amplification systems.
    Figure Legend Snippet: Pearson's Correlation of the signal (MAS5.0) obtained from Affymetrix GeneChips hybridised with cDNA probes synthesised using the NuGen WT-Ovation RNA amplification systems and the effect of reducing RNA input on the resultant Affymetrix GeneChips. ( A ) R 2 values generated when comparing one GeneChip from 50 ng of input HUVEC RNA and duplicate GeneChips from 500 pg and 250 pg of input HUVEC RNA using MAS5.0 analysed data by Pearson's correlation of signal. ( B ) Duplicate GeneChips from 500 pg, 250 pg, 100 pg, 50 pg and 10 pg RNA input compared using MAS5.0 analysed data by Pearson's correlation of signal. C . Principal Component Analysis of all probe sets from GeneChips hybridised with cDNA probes from either WT-Ovation FFPE or WT-Ovation One-Direct RNA amplification systems.

    Techniques Used: Amplification, Generated, Formalin-fixed Paraffin-Embedded

    17) Product Images from "Effects of Secondary Metabolite Extract from Phomopsis occulta on β-Amyloid Aggregation"

    Article Title: Effects of Secondary Metabolite Extract from Phomopsis occulta on β-Amyloid Aggregation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0109438

    Effect of ME0-W-F1 on aggregation of Aβ42 analysed by SDS-PAGE and quantitative analysis with Quantitative One. Low (Aβ42: ME0-W-F1 = 1∶1) and high (Aβ42: ME0-W-F1 = 1∶4) concentrations of ME0-W-F1 were used. Values represent mean ± SD of three replicates. The data were evaluated by t -test, * p
    Figure Legend Snippet: Effect of ME0-W-F1 on aggregation of Aβ42 analysed by SDS-PAGE and quantitative analysis with Quantitative One. Low (Aβ42: ME0-W-F1 = 1∶1) and high (Aβ42: ME0-W-F1 = 1∶4) concentrations of ME0-W-F1 were used. Values represent mean ± SD of three replicates. The data were evaluated by t -test, * p

    Techniques Used: SDS Page

    18) Product Images from "Spliceosome-Mediated Pre-mRNA trans-Splicing Can Repair CEP290 mRNA"

    Article Title: Spliceosome-Mediated Pre-mRNA trans-Splicing Can Repair CEP290 mRNA

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2018.05.014

    Editing of CEP290 Transcripts Occurs in HEK293T after Transfection with PTMs (A) Illustration of adeno-associated virus genome arrangement of a CEP290 5′ PTM with binding domain 07 (PTM_07) with either no poly-adenylation signal (NPA) or with a bovine growth hormone poly-adenylation signal (PA). ITR, inverted terminal repeat; CMV, cytomegalovirus promoter; PCDS, partial coding DNA sequence of CEP290 ; 5′ SS, 5′ splice site. (B) qPCR was performed on HEK293T transfected with a plasmid encoding GFP as a transfection control or the plasmids in (A). TaqMan probes were designed to the junctions indicated: CEP290 exons 26 and 27 (hEx26-hEx27), a region within the 5′ codon optimized partial coding DNA sequence (coPCDS), or the novel junction of 5′ coPCDS and endogenous Homo sapiens exon 27 to signify trans -splicing (coPCDS-hEx27). Significant variation within replicates for coPCDS-hEx27 was present because the Ct was crossed at 35 cycles with either PTM; however, no amplification was observed through 40 cycles in GFP-treated samples. Samples were standardized to β-2-microglobin and normalized to NPA. Error bars are relative quantity minimum and maximum 95% confidence intervals. (C) Agarose gel electrophoresis of one of the replicate reactions from (B). (D) Densitometry analysis of the bands in (C). (E) Sanger sequencing following TOPO-cloning of the PCR product comprising the coPCDS-hEx27 junction visualized in (C). Nucleotide differences between Homo sapiens and codon-optimized CEP290 are noted by asterisks. The junction between the 5′ coPCDS and endogenous exon 27 is marked by a vertical dashed line.
    Figure Legend Snippet: Editing of CEP290 Transcripts Occurs in HEK293T after Transfection with PTMs (A) Illustration of adeno-associated virus genome arrangement of a CEP290 5′ PTM with binding domain 07 (PTM_07) with either no poly-adenylation signal (NPA) or with a bovine growth hormone poly-adenylation signal (PA). ITR, inverted terminal repeat; CMV, cytomegalovirus promoter; PCDS, partial coding DNA sequence of CEP290 ; 5′ SS, 5′ splice site. (B) qPCR was performed on HEK293T transfected with a plasmid encoding GFP as a transfection control or the plasmids in (A). TaqMan probes were designed to the junctions indicated: CEP290 exons 26 and 27 (hEx26-hEx27), a region within the 5′ codon optimized partial coding DNA sequence (coPCDS), or the novel junction of 5′ coPCDS and endogenous Homo sapiens exon 27 to signify trans -splicing (coPCDS-hEx27). Significant variation within replicates for coPCDS-hEx27 was present because the Ct was crossed at 35 cycles with either PTM; however, no amplification was observed through 40 cycles in GFP-treated samples. Samples were standardized to β-2-microglobin and normalized to NPA. Error bars are relative quantity minimum and maximum 95% confidence intervals. (C) Agarose gel electrophoresis of one of the replicate reactions from (B). (D) Densitometry analysis of the bands in (C). (E) Sanger sequencing following TOPO-cloning of the PCR product comprising the coPCDS-hEx27 junction visualized in (C). Nucleotide differences between Homo sapiens and codon-optimized CEP290 are noted by asterisks. The junction between the 5′ coPCDS and endogenous exon 27 is marked by a vertical dashed line.

    Techniques Used: Transfection, Binding Assay, Sequencing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Amplification, Agarose Gel Electrophoresis, Clone Assay, Polymerase Chain Reaction

    Editing of Mini- CEP290 Transcripts Occurs In Vivo following Sub-retinal Injection of 7m8AAV-5′ PTMs (A) Diagram of the CEP290 intron 26 mini-gene. The mini-gene is driven by the murine Rhodopsin promoter (mRho). CEP290 exons 25 and 26 are joined and followed by the complete intron 26-27 and exon 27. An A > G mutation corresponding to c.2991+1655 was included to assess exon X splicing in the model. An amino-terminal Myc and a carboxy-terminal FLAG tag were added to flank the mini-gene. Additionally, an internal ribosomal entry site (IRES) was added to drive expression of EGFP and is terminated by a bovine growth hormone poly-adenylation signal sequence (pA). (B) Immunofluorescence staining of a retinal cross-section from a mini- CEP290 mouse. OS, outer segment. ONL, outer nuclear layer. INL, inner nuclear layer. Scale bar is 50 μm. (C) Illustration of potential splicing outcomes within the mini- CEP290 mouse. Canonical cis- splicing would result in a predicted 24.3-kDa peptide. Alternative cis- splicing with exon X would yield a predicted non-sense media decay transcript encoding a truncated peptide with Myc at 17.1 kDa and no FLAG translation. Trans -splicing with a 5′ PTM would replace the amino-terminal Myc and generate a 124.1-kDa peptide with a FLAG tag. Arrow pairs indicate locations of PCR primers used to detect specific splicing events. (D) Representative western blot images of Myc and FLAG at 24.3 kDa showing OD and OS lanes per animal for each contralateral treatment cohort. (E) Densitometry quantification showing the mean log 10 values of each contralateral treatment cohort. Samples were standardized to α-tubulin. Error bars are SEM. Sample sizes as indicated. Individual animal data is available in Figure S1 A. (F) qPCR from cDNA generated by RNA extracts from whole eyes of mini- CEP290 mice. TaqMan probes were designed to the junctions of Homo sapiens exon 27 and FLAG to detect total expression of the mini-gene (left) to a region within the PCDS to detect total expression of the PTM (center) or to the novel junction of codon-optimized CEP290 PCDS and Homo sapiens CEP290 exon 27 (right). Values from treatment-matched samples were averaged as biological groups, standardized to murine β-2-microglobin and normalized to PTM_NBD. p
    Figure Legend Snippet: Editing of Mini- CEP290 Transcripts Occurs In Vivo following Sub-retinal Injection of 7m8AAV-5′ PTMs (A) Diagram of the CEP290 intron 26 mini-gene. The mini-gene is driven by the murine Rhodopsin promoter (mRho). CEP290 exons 25 and 26 are joined and followed by the complete intron 26-27 and exon 27. An A > G mutation corresponding to c.2991+1655 was included to assess exon X splicing in the model. An amino-terminal Myc and a carboxy-terminal FLAG tag were added to flank the mini-gene. Additionally, an internal ribosomal entry site (IRES) was added to drive expression of EGFP and is terminated by a bovine growth hormone poly-adenylation signal sequence (pA). (B) Immunofluorescence staining of a retinal cross-section from a mini- CEP290 mouse. OS, outer segment. ONL, outer nuclear layer. INL, inner nuclear layer. Scale bar is 50 μm. (C) Illustration of potential splicing outcomes within the mini- CEP290 mouse. Canonical cis- splicing would result in a predicted 24.3-kDa peptide. Alternative cis- splicing with exon X would yield a predicted non-sense media decay transcript encoding a truncated peptide with Myc at 17.1 kDa and no FLAG translation. Trans -splicing with a 5′ PTM would replace the amino-terminal Myc and generate a 124.1-kDa peptide with a FLAG tag. Arrow pairs indicate locations of PCR primers used to detect specific splicing events. (D) Representative western blot images of Myc and FLAG at 24.3 kDa showing OD and OS lanes per animal for each contralateral treatment cohort. (E) Densitometry quantification showing the mean log 10 values of each contralateral treatment cohort. Samples were standardized to α-tubulin. Error bars are SEM. Sample sizes as indicated. Individual animal data is available in Figure S1 A. (F) qPCR from cDNA generated by RNA extracts from whole eyes of mini- CEP290 mice. TaqMan probes were designed to the junctions of Homo sapiens exon 27 and FLAG to detect total expression of the mini-gene (left) to a region within the PCDS to detect total expression of the PTM (center) or to the novel junction of codon-optimized CEP290 PCDS and Homo sapiens CEP290 exon 27 (right). Values from treatment-matched samples were averaged as biological groups, standardized to murine β-2-microglobin and normalized to PTM_NBD. p

    Techniques Used: In Vivo, Injection, Mutagenesis, FLAG-tag, Expressing, Sequencing, Immunofluorescence, Staining, Polymerase Chain Reaction, Western Blot, Real-time Polymerase Chain Reaction, Generated, Mouse Assay

    19) Product Images from "Human Melanocyte-Derived Spheroids: A Precise Test System for Drug Screening and a Multicellular Unit for Tissue Engineering"

    Article Title: Human Melanocyte-Derived Spheroids: A Precise Test System for Drug Screening and a Multicellular Unit for Tissue Engineering

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2020.00540

    Real-time PCR analysis of TYR and MC1R expression in MelanoDerm™ tissue equivalents and spheroids from human melanocytes on Days 1, 3, and 7 in the presence of fucoxanthin and in the control group with growth medium. * p
    Figure Legend Snippet: Real-time PCR analysis of TYR and MC1R expression in MelanoDerm™ tissue equivalents and spheroids from human melanocytes on Days 1, 3, and 7 in the presence of fucoxanthin and in the control group with growth medium. * p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    20) Product Images from "A Single Nucleotide Polymorphism Associated with Hepatitis C Virus Infections Located in the Distal Region of the IL28B Promoter Influences NF-?B-Mediated Gene Transcription"

    Article Title: A Single Nucleotide Polymorphism Associated with Hepatitis C Virus Infections Located in the Distal Region of the IL28B Promoter Influences NF-?B-Mediated Gene Transcription

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0075495

    The distal fragment of the IL28B promoter is responsive to NF-κB. A) A schematic of the 1.4 kb fragment of the IL28B promoter fragment cloned in to pGL3basic vector. The rectangles show the different transcription factor binding sites identified by Osterlund et al. (2007) [21] . The vertical lines next to the non-consensus NF-κB site indicate the position of two SNPs rs28416813 and rs71356849 respectively. ISRE, Interferon-stimulated response element, PRDI, positive-regulatory domain I. The TSS is shown by the arrow before the non-consensus NF-κB-binding site (at position +115 with respect to TSS). B) Luciferase assays carried out with HEK293T cells in 6-well plates. The transfection of the p1.4IL28BC-T construct was done either in presence or absence of p50+p65 and activity compared with the empty pGL3basic vector. The error bars show SD from triplicates.
    Figure Legend Snippet: The distal fragment of the IL28B promoter is responsive to NF-κB. A) A schematic of the 1.4 kb fragment of the IL28B promoter fragment cloned in to pGL3basic vector. The rectangles show the different transcription factor binding sites identified by Osterlund et al. (2007) [21] . The vertical lines next to the non-consensus NF-κB site indicate the position of two SNPs rs28416813 and rs71356849 respectively. ISRE, Interferon-stimulated response element, PRDI, positive-regulatory domain I. The TSS is shown by the arrow before the non-consensus NF-κB-binding site (at position +115 with respect to TSS). B) Luciferase assays carried out with HEK293T cells in 6-well plates. The transfection of the p1.4IL28BC-T construct was done either in presence or absence of p50+p65 and activity compared with the empty pGL3basic vector. The error bars show SD from triplicates.

    Techniques Used: Clone Assay, Plasmid Preparation, Binding Assay, Luciferase, Transfection, Construct, Activity Assay

    21) Product Images from "Genetic and Epigenetic Profiling Reveals EZH2-mediated Down Regulation of OCT-4 Involves NR2F2 during Cardiac Differentiation of Human Embryonic Stem Cells"

    Article Title: Genetic and Epigenetic Profiling Reveals EZH2-mediated Down Regulation of OCT-4 Involves NR2F2 during Cardiac Differentiation of Human Embryonic Stem Cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-13442-9

    Distribution of H3K27me3 mark on OCT4. Binding profile of H3K27me3 mark on the OCT4 gene during cardiac differentiation generated by IGV genome browser mapped to human hg 19 genome.
    Figure Legend Snippet: Distribution of H3K27me3 mark on OCT4. Binding profile of H3K27me3 mark on the OCT4 gene during cardiac differentiation generated by IGV genome browser mapped to human hg 19 genome.

    Techniques Used: Binding Assay, Generated

    Distribution of H3K27me3 mark on NR2F2. Occupancy of H3K27me3 mark on the NR2F2 locus at days 0, 12 and 20 generated by IGV gene browser mapped to human hg 19 genome.
    Figure Legend Snippet: Distribution of H3K27me3 mark on NR2F2. Occupancy of H3K27me3 mark on the NR2F2 locus at days 0, 12 and 20 generated by IGV gene browser mapped to human hg 19 genome.

    Techniques Used: Generated

    22) Product Images from "Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells"

    Article Title: Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.6318

    Optical density values of K562 cells following 48 h drug-treatment with (A) DAC (EC 50 , 6457 nM), (B) nilotinib (EC 50 , 10.55 nM) and (C) IM (EC 50 , 227.5 nM) were measured using a Cell Counting Kit-8. (D) K562 cells treated with 5 µM DAC combined with 0.1 µM IM or 5 nM nilotinib exhibited proliferation inhibition that was statistically significant compared with the IM or nilotinib monotherapy treatment groups. ***P
    Figure Legend Snippet: Optical density values of K562 cells following 48 h drug-treatment with (A) DAC (EC 50 , 6457 nM), (B) nilotinib (EC 50 , 10.55 nM) and (C) IM (EC 50 , 227.5 nM) were measured using a Cell Counting Kit-8. (D) K562 cells treated with 5 µM DAC combined with 0.1 µM IM or 5 nM nilotinib exhibited proliferation inhibition that was statistically significant compared with the IM or nilotinib monotherapy treatment groups. ***P

    Techniques Used: Cell Counting, Inhibition

    23) Product Images from "Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells"

    Article Title: Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.6318

    Optical density values of K562 cells following 48 h drug-treatment with (A) DAC (EC 50 , 6457 nM), (B) nilotinib (EC 50 , 10.55 nM) and (C) IM (EC 50 , 227.5 nM) were measured using a Cell Counting Kit-8. (D) K562 cells treated with 5 µM DAC combined with 0.1 µM IM or 5 nM nilotinib exhibited proliferation inhibition that was statistically significant compared with the IM or nilotinib monotherapy treatment groups. ***P
    Figure Legend Snippet: Optical density values of K562 cells following 48 h drug-treatment with (A) DAC (EC 50 , 6457 nM), (B) nilotinib (EC 50 , 10.55 nM) and (C) IM (EC 50 , 227.5 nM) were measured using a Cell Counting Kit-8. (D) K562 cells treated with 5 µM DAC combined with 0.1 µM IM or 5 nM nilotinib exhibited proliferation inhibition that was statistically significant compared with the IM or nilotinib monotherapy treatment groups. ***P

    Techniques Used: Cell Counting, Inhibition

    24) Product Images from "LSD1 dual function in mediating epigenetic corruption of the vitamin D signaling in prostate cancer"

    Article Title: LSD1 dual function in mediating epigenetic corruption of the vitamin D signaling in prostate cancer

    Journal: Clinical Epigenetics

    doi: 10.1186/s13148-017-0382-y

    a Immunoprecipitation (IP) and western blotting (WB) data showing that LSD1 and VDR belong to the same transcriptional complex. IP was performed from nuclear lysate in samples treated with vehicle control or 1,25-D 3 using the same LSD1 antibody described for IHC and WB. In post-IP, the samples were probed for LSD1 and VDR. The same double band visible in Fig. 1 was also detected in this sample. b , c Effect of LSD1 knockdown and 1,25-D 3 treatment on gene expression of VDR target genes. Every graph compares the effect of vitamin D in control (CTR) cells and LSD1 knockdown (siLSD1) cells. Each bar is the mean of at least three biological replicates with SEM, showing the fold changes of treated (+ D3) vs. vehicle-treated (− Veh) samples. The columns indicate, from left to right , siCTR + Veh, siCTR + 1,25-D 3 , siLSD1 + Veh, and siLSD1 + 1,25-D 3 . Transcript levels were measured for b E2f1 and Cdkn1a and c Cyp24a1 and S100g. Statistical significance was evaluated with one-way ANOVA and Tukey post hoc correction (*** p
    Figure Legend Snippet: a Immunoprecipitation (IP) and western blotting (WB) data showing that LSD1 and VDR belong to the same transcriptional complex. IP was performed from nuclear lysate in samples treated with vehicle control or 1,25-D 3 using the same LSD1 antibody described for IHC and WB. In post-IP, the samples were probed for LSD1 and VDR. The same double band visible in Fig. 1 was also detected in this sample. b , c Effect of LSD1 knockdown and 1,25-D 3 treatment on gene expression of VDR target genes. Every graph compares the effect of vitamin D in control (CTR) cells and LSD1 knockdown (siLSD1) cells. Each bar is the mean of at least three biological replicates with SEM, showing the fold changes of treated (+ D3) vs. vehicle-treated (− Veh) samples. The columns indicate, from left to right , siCTR + Veh, siCTR + 1,25-D 3 , siLSD1 + Veh, and siLSD1 + 1,25-D 3 . Transcript levels were measured for b E2f1 and Cdkn1a and c Cyp24a1 and S100g. Statistical significance was evaluated with one-way ANOVA and Tukey post hoc correction (*** p

    Techniques Used: Immunoprecipitation, Western Blot, Immunohistochemistry, Expressing

    ChIP analysis of BC1A cells stably transfected with shLSD1 lentiviral vector and treated for 24 h with 100 nM 1,25-D 3 . Each bar indicates the percentage of binding relative to INPUT and represents the mean of at least three biological replicates with SEM. The columns indicate, from left to right , shCTR + Veh, shCTR + 1,25-D 3 , shLSD1 + Veh, and shLSD1 + 1,25-D 3 . The basal levels of the following protein/histone marks were evaluated, from left to right , in each graph: IgG , VDR , DNMT1 , H3K4me2 , and H3K9Ac . The regions analyzed were a Cdkn1a TSS, b Cdkn1a VDRE, c E2f1 TSS, d Cyp24a1 TSS, and e S100g TSS. IgG was used as a control for non-specific binding/enrichment. Statistical significance was calculated using one-way ANOVA and Tukey post hoc correction (*** p
    Figure Legend Snippet: ChIP analysis of BC1A cells stably transfected with shLSD1 lentiviral vector and treated for 24 h with 100 nM 1,25-D 3 . Each bar indicates the percentage of binding relative to INPUT and represents the mean of at least three biological replicates with SEM. The columns indicate, from left to right , shCTR + Veh, shCTR + 1,25-D 3 , shLSD1 + Veh, and shLSD1 + 1,25-D 3 . The basal levels of the following protein/histone marks were evaluated, from left to right , in each graph: IgG , VDR , DNMT1 , H3K4me2 , and H3K9Ac . The regions analyzed were a Cdkn1a TSS, b Cdkn1a VDRE, c E2f1 TSS, d Cyp24a1 TSS, and e S100g TSS. IgG was used as a control for non-specific binding/enrichment. Statistical significance was calculated using one-way ANOVA and Tukey post hoc correction (*** p

    Techniques Used: Chromatin Immunoprecipitation, Stable Transfection, Transfection, Plasmid Preparation, Binding Assay

    25) Product Images from "Amino acid deprivation triggers a novel GCN2-independent response leading to the transcriptional reactivation of non-native DNA sequences"

    Article Title: Amino acid deprivation triggers a novel GCN2-independent response leading to the transcriptional reactivation of non-native DNA sequences

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0200783

    Transgene reactivation is abolished by MAPK inhibitors, and is induced by ribosomal inhibitors. (A) Relative transgene (OA1) mRNA abundance in HeLa-OA1 and HepG2-OA1 cells cultured in full medium or Met/Cys-deprived medium, in the presence or absence of inhibitors for MEK1/2 (U0126; 50 μM), and JNK1/2/3 (SP600125; 20–50 μM) for 24 h. For HeLa-OA1 cells data represent the mean ± SEM of 4 (CTRL and SP600125), or the mean ± range of 2 (U0126) independent experiments; for HepG2-OA1 cells, data represent the mean ± SEM of 3 independent experiments. Results are expressed as fold change vs. control (full medium = 1). * P
    Figure Legend Snippet: Transgene reactivation is abolished by MAPK inhibitors, and is induced by ribosomal inhibitors. (A) Relative transgene (OA1) mRNA abundance in HeLa-OA1 and HepG2-OA1 cells cultured in full medium or Met/Cys-deprived medium, in the presence or absence of inhibitors for MEK1/2 (U0126; 50 μM), and JNK1/2/3 (SP600125; 20–50 μM) for 24 h. For HeLa-OA1 cells data represent the mean ± SEM of 4 (CTRL and SP600125), or the mean ± range of 2 (U0126) independent experiments; for HepG2-OA1 cells, data represent the mean ± SEM of 3 independent experiments. Results are expressed as fold change vs. control (full medium = 1). * P

    Techniques Used: Cell Culture

    EAA deprivation induces reactivation of silent transgenes in HeLa and HepG2 cells. Relative transgene (OA1) and CHOP mRNA abundance in HeLa-OA1 (A) and HepG2-OA1 (B) cells, cultured in various AA-deprived media for 48 h and 24 h, respectively, compared to full medium. Mean ± SEM of 3 independent experiments. Data are expressed as fold change vs. control (full medium = 1). *P
    Figure Legend Snippet: EAA deprivation induces reactivation of silent transgenes in HeLa and HepG2 cells. Relative transgene (OA1) and CHOP mRNA abundance in HeLa-OA1 (A) and HepG2-OA1 (B) cells, cultured in various AA-deprived media for 48 h and 24 h, respectively, compared to full medium. Mean ± SEM of 3 independent experiments. Data are expressed as fold change vs. control (full medium = 1). *P

    Techniques Used: Cell Culture

    mTOR inhibition and GCN2 activation differently affect transgene expression in HeLa and HepG2 cells. Relative transgene (OA1) and CHOP mRNA abundance in HeLa-OA1 (A) and HepG2-OA1 (B) cells, cultured in Met/Cys-deprived medium, or in the presence of PP242 (mTOR inhibitor; 1–3 μM) or L-Histidinol (HisOH, GCN2 activator; 4–16 mM), either alone or in combination for 24–48 h, compared to full medium. Mean ± SEM of 4 (A) or 3 (B) independent experiments. Data are expressed as fold change vs. control (full medium = 1). *P
    Figure Legend Snippet: mTOR inhibition and GCN2 activation differently affect transgene expression in HeLa and HepG2 cells. Relative transgene (OA1) and CHOP mRNA abundance in HeLa-OA1 (A) and HepG2-OA1 (B) cells, cultured in Met/Cys-deprived medium, or in the presence of PP242 (mTOR inhibitor; 1–3 μM) or L-Histidinol (HisOH, GCN2 activator; 4–16 mM), either alone or in combination for 24–48 h, compared to full medium. Mean ± SEM of 4 (A) or 3 (B) independent experiments. Data are expressed as fold change vs. control (full medium = 1). *P

    Techniques Used: Inhibition, Activation Assay, Expressing, Cell Culture

    26) Product Images from "A20 deubiquitinase controls PGC-1α expression in the adipose tissue"

    Article Title: A20 deubiquitinase controls PGC-1α expression in the adipose tissue

    Journal: Lipids in Health and Disease

    doi: 10.1186/s12944-018-0740-6

    Inhibiting A20 expression. Obese Swiss mice were treated once a day, for 7 days, with a single intraperitoneal injection of a 100 μl solution containing either A20 antisense (ASO) or scrambled (SCR) oligonucleotide. Fragments from subcutaneous (SC) ( a-d ), visceral (VI) ( e-g ) or brown (BAT) ( h-j ) adipose tissue were employed in quantitative real-time PCR experiments. In a , A20 transcript expression was evaluated in a dose-response experiment to determine the efficiency of the method. In b-l , the doses of ASO and SCR employed were always 2 nmol in 100 μl/dose. In b-j , the expressions of A20 ( b , e , h ); PGC-1α ( c , f , i ); and PPARγ ( d , g , j ) were determined and are represented graphically. The blood glucose levels during a glucose tolerance test ( k ) and the respective area under the glucose curve ( l ) are represented graphically. In all experiments n = 8; * p
    Figure Legend Snippet: Inhibiting A20 expression. Obese Swiss mice were treated once a day, for 7 days, with a single intraperitoneal injection of a 100 μl solution containing either A20 antisense (ASO) or scrambled (SCR) oligonucleotide. Fragments from subcutaneous (SC) ( a-d ), visceral (VI) ( e-g ) or brown (BAT) ( h-j ) adipose tissue were employed in quantitative real-time PCR experiments. In a , A20 transcript expression was evaluated in a dose-response experiment to determine the efficiency of the method. In b-l , the doses of ASO and SCR employed were always 2 nmol in 100 μl/dose. In b-j , the expressions of A20 ( b , e , h ); PGC-1α ( c , f , i ); and PPARγ ( d , g , j ) were determined and are represented graphically. The blood glucose levels during a glucose tolerance test ( k ) and the respective area under the glucose curve ( l ) are represented graphically. In all experiments n = 8; * p

    Techniques Used: Expressing, Mouse Assay, Injection, Allele-specific Oligonucleotide, Real-time Polymerase Chain Reaction, Pyrolysis Gas Chromatography

    27) Product Images from "The grapevine VvibZIPC22 transcription factor is involved in the regulation of flavonoid biosynthesis"

    Article Title: The grapevine VvibZIPC22 transcription factor is involved in the regulation of flavonoid biosynthesis

    Journal: Journal of Experimental Botany

    doi: 10.1093/jxb/erw181

    Functional characterization of VvibZIPC22 in tobacco overexpressing lines. (A) Stamens and corollas of wild-type (WT) plants and transgenic lines (L), and (B) average VvibZIPC22 relative expression in stamens (S) and petal limbs (P) of three WT and six L at two developmental stages (S1=open flower without pollen, S2= mature flower). (C) Variation of cyanidin 3-rutinoside (Cyan 3-rut), quercetin 3-rutinoside (Que 3-rut), and kaempferol 3-rutinoside (Kaempf 3-rut) in stamens and of proanthocyanidins (PAs) in petal limbs of L and WT plants at the two stages. Anthocyanin and flavonol contents were determined by HPLC-DAD, while PA content was determined by DMACA. (D) Fold induction in gene expression of tobacco flavonoid structural genes between L and WT stamens and petal limbs. Transcript levels in both (B) and (D) were determined by qRT-PCR using gene-specific primers ( Supplementary Table S1 ), calibration against the average expression value in all the lines at each stage, and normalization against NteIF4a and NtUbiquitin relative expression. Average values with error bars indicating the SE are shown. Asterisks indicate significant changes ( P
    Figure Legend Snippet: Functional characterization of VvibZIPC22 in tobacco overexpressing lines. (A) Stamens and corollas of wild-type (WT) plants and transgenic lines (L), and (B) average VvibZIPC22 relative expression in stamens (S) and petal limbs (P) of three WT and six L at two developmental stages (S1=open flower without pollen, S2= mature flower). (C) Variation of cyanidin 3-rutinoside (Cyan 3-rut), quercetin 3-rutinoside (Que 3-rut), and kaempferol 3-rutinoside (Kaempf 3-rut) in stamens and of proanthocyanidins (PAs) in petal limbs of L and WT plants at the two stages. Anthocyanin and flavonol contents were determined by HPLC-DAD, while PA content was determined by DMACA. (D) Fold induction in gene expression of tobacco flavonoid structural genes between L and WT stamens and petal limbs. Transcript levels in both (B) and (D) were determined by qRT-PCR using gene-specific primers ( Supplementary Table S1 ), calibration against the average expression value in all the lines at each stage, and normalization against NteIF4a and NtUbiquitin relative expression. Average values with error bars indicating the SE are shown. Asterisks indicate significant changes ( P

    Techniques Used: Functional Assay, Transgenic Assay, Expressing, High Performance Liquid Chromatography, Quantitative RT-PCR

    (A) Profiles of VvibZIPC22 relative expression and of six flavonol aglycons at different Pinot Noir berry developmental stages. (B) VvibZIPC22 relative expression in a panel of 15 Pinot Noir organs. Transcript levels (NRQs) were determined by qRT-PCR using gene-specific primers ( Supplementary Table S1 ), calibration against the mean expression value in all the stages, and normalization against VviGADPH and VviACTIN relative expression. Flavonol content was determined by HPLC-DAD analysis. Each value corresponds to the mean and SE of three different biological replicates. F, flowering (50% opened flowers, E-L 23); P, pepper corn (E-L 29); PV, pre-véraison (hard green berries, E-L 33); V, véraison (50% coloured berries, E-L 35); 16°, post-véraison (berries at 16° Brix, E-L 36); 18°, maturity (berries at 18° Brix, E-L 38); NRQ, normalized relative quantity; FW, fresh weight.
    Figure Legend Snippet: (A) Profiles of VvibZIPC22 relative expression and of six flavonol aglycons at different Pinot Noir berry developmental stages. (B) VvibZIPC22 relative expression in a panel of 15 Pinot Noir organs. Transcript levels (NRQs) were determined by qRT-PCR using gene-specific primers ( Supplementary Table S1 ), calibration against the mean expression value in all the stages, and normalization against VviGADPH and VviACTIN relative expression. Flavonol content was determined by HPLC-DAD analysis. Each value corresponds to the mean and SE of three different biological replicates. F, flowering (50% opened flowers, E-L 23); P, pepper corn (E-L 29); PV, pre-véraison (hard green berries, E-L 33); V, véraison (50% coloured berries, E-L 35); 16°, post-véraison (berries at 16° Brix, E-L 36); 18°, maturity (berries at 18° Brix, E-L 38); NRQ, normalized relative quantity; FW, fresh weight.

    Techniques Used: Expressing, Quantitative RT-PCR, High Performance Liquid Chromatography

    UV light responsiveness of VvibZIPC22 and flavonol pathway genes. (A) Induction of VvibZIPC22 , VviFLS1 , and VviMYBF1 in Chardonnay leaves after UV light treatment. Each bar corresponds to the fold induction in gene expression between treated and control leaves as determined by qRT-PCR. Transcript levels were measured using gene-specific primers ( Supplementary Table S1 ), calibration against the expression value in the control sample at 0h post-treatment, and normalization against VviGADPH and VviUbiquitin relative expression. (B) Accumulation of flavonols in Chardonnay leaves after UV light treatment. Each bar corresponds to the fold induction in the total content of flavonol aglycons between treated and control leaves as detected by HPLC-DAD analysis. For both measurements, each data point corresponds to the mean and SE of three independent extractions of the same biological material. (C) Putative light regulatory elements identified in the promoter of VvibZIPC22 (PN, Ch) in this study (underlined) and in the promoters of VviMYBF1 and VviFLS1 ( Czemmel et al. , 2009 ). These cis -elements were aligned with the corresponding Arabidopsis elements taken from the PLACE database ( Higo et al. , 1999 ) using GeneDoc software (v. 1.6). Nucleotides are labelled from black to light grey according to their conservation. Numbers in front of the alignment indicate the relative distance of each element from the putative transcriptional start site (+1). CHS, chalcone synthase; FLS, flavonol synthase; PN, Pinot Noir; Ch, Chardonnay; ACS, ACGT-containing sequence similar to ACE in the Arabidopsis CHS promoter (accession no. S000355); MRS, MYB recognition sequence similar to MRE in the Arabidopsis CHS promoter (S000356); MYBCORE (S000176), IBOXCORE element (S000199); RRS, R response sequence similar to RRE in the Arabidopsis CHS promoter (S000407).
    Figure Legend Snippet: UV light responsiveness of VvibZIPC22 and flavonol pathway genes. (A) Induction of VvibZIPC22 , VviFLS1 , and VviMYBF1 in Chardonnay leaves after UV light treatment. Each bar corresponds to the fold induction in gene expression between treated and control leaves as determined by qRT-PCR. Transcript levels were measured using gene-specific primers ( Supplementary Table S1 ), calibration against the expression value in the control sample at 0h post-treatment, and normalization against VviGADPH and VviUbiquitin relative expression. (B) Accumulation of flavonols in Chardonnay leaves after UV light treatment. Each bar corresponds to the fold induction in the total content of flavonol aglycons between treated and control leaves as detected by HPLC-DAD analysis. For both measurements, each data point corresponds to the mean and SE of three independent extractions of the same biological material. (C) Putative light regulatory elements identified in the promoter of VvibZIPC22 (PN, Ch) in this study (underlined) and in the promoters of VviMYBF1 and VviFLS1 ( Czemmel et al. , 2009 ). These cis -elements were aligned with the corresponding Arabidopsis elements taken from the PLACE database ( Higo et al. , 1999 ) using GeneDoc software (v. 1.6). Nucleotides are labelled from black to light grey according to their conservation. Numbers in front of the alignment indicate the relative distance of each element from the putative transcriptional start site (+1). CHS, chalcone synthase; FLS, flavonol synthase; PN, Pinot Noir; Ch, Chardonnay; ACS, ACGT-containing sequence similar to ACE in the Arabidopsis CHS promoter (accession no. S000355); MRS, MYB recognition sequence similar to MRE in the Arabidopsis CHS promoter (S000356); MYBCORE (S000176), IBOXCORE element (S000199); RRS, R response sequence similar to RRE in the Arabidopsis CHS promoter (S000407).

    Techniques Used: Expressing, Quantitative RT-PCR, High Performance Liquid Chromatography, Software, Sequencing

    28) Product Images from "Edwardsiella tarda Outer Membrane Protein C: An Immunogenic Protein Induces Highly Protective Effects in Flounder (Paralichthys olivaceus) against Edwardsiellosis"

    Article Title: Edwardsiella tarda Outer Membrane Protein C: An Immunogenic Protein Induces Highly Protective Effects in Flounder (Paralichthys olivaceus) against Edwardsiellosis

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms17071117

    qRT-PCR analysis of the expression of MHCIIα ( A ) and CD4-1 ( B ). The liver, spleen, and head kidney were sampled at 1, 2, 3, 5, 7, 14, 21 and 28 days after immunization. The mRNA level of each immune-related gene was normalized to that of 18S . For each gene, the mRNA level of the control fish was set as 1. Results are expressed as means ± SEM ( n = 3). The asterisk indicates the statistical significance compared with the control group (* p
    Figure Legend Snippet: qRT-PCR analysis of the expression of MHCIIα ( A ) and CD4-1 ( B ). The liver, spleen, and head kidney were sampled at 1, 2, 3, 5, 7, 14, 21 and 28 days after immunization. The mRNA level of each immune-related gene was normalized to that of 18S . For each gene, the mRNA level of the control fish was set as 1. Results are expressed as means ± SEM ( n = 3). The asterisk indicates the statistical significance compared with the control group (* p

    Techniques Used: Quantitative RT-PCR, Expressing, Fluorescence In Situ Hybridization

    qRT-PCR analysis of the expression of IL-1β ( A ), TNF-α ( B ) and IL-6 ( C ). The liver, spleen, and head kidney were sampled at 6 h, 12 h, 24 h, 48 h, 96 h, 5 days and 7 days after immunization. The mRNA levels of each immune-related gene were normalized to those of 18S . For each gene, the mRNA level of the control fish was set as 1. Results are expressed as means ± SEM ( n = 3). The asterisk indicates the statistical significance compared with the control group (* p
    Figure Legend Snippet: qRT-PCR analysis of the expression of IL-1β ( A ), TNF-α ( B ) and IL-6 ( C ). The liver, spleen, and head kidney were sampled at 6 h, 12 h, 24 h, 48 h, 96 h, 5 days and 7 days after immunization. The mRNA levels of each immune-related gene were normalized to those of 18S . For each gene, the mRNA level of the control fish was set as 1. Results are expressed as means ± SEM ( n = 3). The asterisk indicates the statistical significance compared with the control group (* p

    Techniques Used: Quantitative RT-PCR, Expressing, Fluorescence In Situ Hybridization

    29) Product Images from "Stemness and differentiation potential-recovery effects of sinapic acid against ultraviolet-A-induced damage through the regulation of p38 MAPK and NF-κB"

    Article Title: Stemness and differentiation potential-recovery effects of sinapic acid against ultraviolet-A-induced damage through the regulation of p38 MAPK and NF-κB

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-01089-5

    Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF gene expression. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) Intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. ( B ) The triglyceride content was measured using a triglyceride assay kit (Cayman Chemical, Ann Arbor, MI). The results were verified by three repetitions of the experiments, each of which was conducted in triplicate. ( C ) At day 14 after the induction of differentiation, the supernatants were harvested for MIF measurement. Data are expressed as the means ± S.D. ( D ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the MIF gene were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. * p
    Figure Legend Snippet: Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF gene expression. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) Intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. ( B ) The triglyceride content was measured using a triglyceride assay kit (Cayman Chemical, Ann Arbor, MI). The results were verified by three repetitions of the experiments, each of which was conducted in triplicate. ( C ) At day 14 after the induction of differentiation, the supernatants were harvested for MIF measurement. Data are expressed as the means ± S.D. ( D ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the MIF gene were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. * p

    Techniques Used: Expressing, Irradiation, Staining, Isolation, Quantitative RT-PCR

    Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF-AMPK-KLF2 signaling by inhibiting NF-κB. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) At 14 days after the induction of differentiation, the total lysates were analyzed by Western blot using the indicated antibodies. ( B ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the KLF2 and PPARγ genes were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. ( C ) At 1 h after the induction of differentiation, cell lysates were analyzed using a Multi-Target Sandwich ELISA Kit. All of the results were verified by repeating the experiments, each of which was conducted in duplicate, three times. SA: sinapic acid, DIF: differentiation media, UVA: ultraviolet A. Data are expressed as the means ± S.D. * p
    Figure Legend Snippet: Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF-AMPK-KLF2 signaling by inhibiting NF-κB. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) At 14 days after the induction of differentiation, the total lysates were analyzed by Western blot using the indicated antibodies. ( B ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the KLF2 and PPARγ genes were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. ( C ) At 1 h after the induction of differentiation, cell lysates were analyzed using a Multi-Target Sandwich ELISA Kit. All of the results were verified by repeating the experiments, each of which was conducted in duplicate, three times. SA: sinapic acid, DIF: differentiation media, UVA: ultraviolet A. Data are expressed as the means ± S.D. * p

    Techniques Used: Irradiation, Western Blot, Isolation, Quantitative RT-PCR, Sandwich ELISA

    30) Product Images from "Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells"

    Article Title: Role and mechanism of decitabine combined with tyrosine kinase inhibitors in advanced chronic myeloid leukemia cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.6318

    RT-qPCR analysis of SHP-1 and BCR/ABL mRNA expression. (A) SHP-1 and BCR/ABL mRNA expression in K562 cells following 48 h drug-treated through RT-qPCR analysis. Expression of (B) SHP-1 and (C) BCR/ABL mRNA expression in chronic myeloid leukemia-chronic phase mononuclear cells following 48 h drug treatment through RT-qPCR analysis. *P
    Figure Legend Snippet: RT-qPCR analysis of SHP-1 and BCR/ABL mRNA expression. (A) SHP-1 and BCR/ABL mRNA expression in K562 cells following 48 h drug-treated through RT-qPCR analysis. Expression of (B) SHP-1 and (C) BCR/ABL mRNA expression in chronic myeloid leukemia-chronic phase mononuclear cells following 48 h drug treatment through RT-qPCR analysis. *P

    Techniques Used: Quantitative RT-PCR, Expressing

    31) Product Images from "Establishment of leukemia inhibitory factor (LIF)-independent iPS cells with potentiated Oct4"

    Article Title: Establishment of leukemia inhibitory factor (LIF)-independent iPS cells with potentiated Oct4

    Journal: Stem cell research

    doi: 10.1016/j.scr.2015.09.002

    Comparison of global expression patterns of mRNA in iPSCs, ESCs, and MEFs
    Figure Legend Snippet: Comparison of global expression patterns of mRNA in iPSCs, ESCs, and MEFs

    Techniques Used: Expressing

    Expression levels of selected miRNAs in MEFs, ESCs, and iPSCs
    Figure Legend Snippet: Expression levels of selected miRNAs in MEFs, ESCs, and iPSCs

    Techniques Used: Expressing

    32) Product Images from "Role of miR-27a, miR-181a and miR-20b in gastric cancer hypoxia-induced chemoresistance"

    Article Title: Role of miR-27a, miR-181a and miR-20b in gastric cancer hypoxia-induced chemoresistance

    Journal: Cancer Biology & Therapy

    doi: 10.1080/15384047.2016.1139244

    Median levels ofmiR-27a, miR-181a and miR-20b in GC patients with PD vs those with DCR (A). Percentage of miR-27a, miR-181a and miR-20b overexpression in GC patients with PD vs those with DCR (B). Median levels of HIF1A, MDR1 and HIPK2 in GC patients
    Figure Legend Snippet: Median levels ofmiR-27a, miR-181a and miR-20b in GC patients with PD vs those with DCR (A). Percentage of miR-27a, miR-181a and miR-20b overexpression in GC patients with PD vs those with DCR (B). Median levels of HIF1A, MDR1 and HIPK2 in GC patients

    Techniques Used: Over Expression

    Correlation analysis between HIF1A and miR-20bexpression (A) MDR1 and miR-27a levels (B) and HIPK2 and miR-181a (C) and HIPK2 and miR-27a (D) in GC patients.
    Figure Legend Snippet: Correlation analysis between HIF1A and miR-20bexpression (A) MDR1 and miR-27a levels (B) and HIPK2 and miR-181a (C) and HIPK2 and miR-27a (D) in GC patients.

    Techniques Used:

    33) Product Images from "Effects of Bni5 Binding on Septin Filament Organization"

    Article Title: Effects of Bni5 Binding on Septin Filament Organization

    Journal: Journal of molecular biology

    doi: 10.1016/j.jmb.2016.10.024

    FRET analysis of Bni5-septin filament interaction. (a) Coomassie and Typhoon scans of Bni5 C144 AF647 ( upper left ), Bni5 C266 AF647 ( upper right ), Bni5 C375 AF647 ( lower left ) and Cdc11 AF555 -capped septin hetero-octamers ( lower right ). Black arrows , labeled protein; gray arrows , degradation products. (b) Maximum efficFigiency of transfer (FRET signal) at an excess concentration of the indicated single-Cys labeled Bni5 AF647 protein (500 nM) with filaments composed of Cdc11 AF555 septin hetero-octamers (25 nM). (c) Coomassie and Typhoon scans of Bni5(1–313) C144 AF647 ( upper left) , Bni5(314–448) C375 AF647 ( upper right ) Cdc11(ΔCTE) AF555 capped hetero-octamers ( lower left ) and Cdc11(Δα0) AF555 capped hetero-octamers. Black arrows , labeled protein. (d) Maximum efficiency of transfer (FRET signal) at an excess concentration of the indicated single-Cys labeled Bni5 AF647 protein (500 nM) with filaments composed of the indicated Cdc11 AF555 septin hetero-octamer (25 nM). Asterisks , for all three types of hetero-octamers [Cdc11-, Cdc11(ΔCTE)- and Cdc11(Δα0)-capped], the reduction in FRET observed for Bni5(1–313) was statistically significant (p
    Figure Legend Snippet: FRET analysis of Bni5-septin filament interaction. (a) Coomassie and Typhoon scans of Bni5 C144 AF647 ( upper left ), Bni5 C266 AF647 ( upper right ), Bni5 C375 AF647 ( lower left ) and Cdc11 AF555 -capped septin hetero-octamers ( lower right ). Black arrows , labeled protein; gray arrows , degradation products. (b) Maximum efficFigiency of transfer (FRET signal) at an excess concentration of the indicated single-Cys labeled Bni5 AF647 protein (500 nM) with filaments composed of Cdc11 AF555 septin hetero-octamers (25 nM). (c) Coomassie and Typhoon scans of Bni5(1–313) C144 AF647 ( upper left) , Bni5(314–448) C375 AF647 ( upper right ) Cdc11(ΔCTE) AF555 capped hetero-octamers ( lower left ) and Cdc11(Δα0) AF555 capped hetero-octamers. Black arrows , labeled protein. (d) Maximum efficiency of transfer (FRET signal) at an excess concentration of the indicated single-Cys labeled Bni5 AF647 protein (500 nM) with filaments composed of the indicated Cdc11 AF555 septin hetero-octamer (25 nM). Asterisks , for all three types of hetero-octamers [Cdc11-, Cdc11(ΔCTE)- and Cdc11(Δα0)-capped], the reduction in FRET observed for Bni5(1–313) was statistically significant (p

    Techniques Used: Labeling, Concentration Assay

    34) Product Images from "Development and Validation of Burkholderia pseudomallei-Specific Real-Time PCR Assays for Clinical, Environmental or Forensic Detection Applications"

    Article Title: Development and Validation of Burkholderia pseudomallei-Specific Real-Time PCR Assays for Clinical, Environmental or Forensic Detection Applications

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0037723

    122018 TaqMan dual probe assay. Red, the B. pseudomallei- specific TaqMan probe amplifies only B. pseudomallei template; the non- B.pseudomallei TaqMan probe (green) amplifies well with B. thailandensis and B. thailandensis- like species and weakly with B. oklahomensis templates but not B. pseudomallei . Other Burkholderia spp. do not amplify with either probe.
    Figure Legend Snippet: 122018 TaqMan dual probe assay. Red, the B. pseudomallei- specific TaqMan probe amplifies only B. pseudomallei template; the non- B.pseudomallei TaqMan probe (green) amplifies well with B. thailandensis and B. thailandensis- like species and weakly with B. oklahomensis templates but not B. pseudomallei . Other Burkholderia spp. do not amplify with either probe.

    Techniques Used:

    266152 TaqMan dual probe assay. Green, the B. pseudomallei -specific TaqMan probe preferentially amplifies B. pseudomallei template; the non- B.pseudomallei TaqMan probe (red) amplifies well in B. thailandensis- like species and weakly in B. thailandensis and B. oklahomensis . Other Burkholderia spp. do not amplify.
    Figure Legend Snippet: 266152 TaqMan dual probe assay. Green, the B. pseudomallei -specific TaqMan probe preferentially amplifies B. pseudomallei template; the non- B.pseudomallei TaqMan probe (red) amplifies well in B. thailandensis- like species and weakly in B. thailandensis and B. oklahomensis . Other Burkholderia spp. do not amplify.

    Techniques Used:

    35) Product Images from "Modulation of human embryonic stem cell-derived cardiomyocyte growth: A testbed for studying human cardiac hypertrophy?"

    Article Title: Modulation of human embryonic stem cell-derived cardiomyocyte growth: A testbed for studying human cardiac hypertrophy?

    Journal: Journal of Molecular and Cellular Cardiology

    doi: 10.1016/j.yjmcc.2010.10.029

    Phenylephrine modulates cell size independently of cell cycle. (A) Representative immunofluorescence image showing hESC-CM stained positive for the myosin heavy chain α/β (MHCα/β, green), DAPI (blue), Ki67 (nuclear, red), and atrial natriuretic factor (ANF, perinuclear, orange) in the presence of cell cycle inhibitor blebbistatin (10 μM) at 30 days after differentiation. Scale bar represents 50 μm. Bar graphs showing cell size of hESC-CM (B), percentage of binucleated hESC-CM (C), and percentage of Ki67-positive hESC-CM (D) treated with phenylephrine (PE) in the presence of blebbistatin (Bleb, solid bar) or vehicle (light grey bar). Results are shown as mean ± SEM ( n > 100 MHC-positive cells per well, in triplicate, n = 2 preparations). The Cellomics Cell Cycle BioApplication classified hESC-CM treated with blebbistatin, nocodazole, or control medium into their cell cycle phase based on the total nuclear intensity of DNA binding DAPI (E). Cell cycle distribution presented as histogram where the Y -axis represents the number of instances and the X -axis represents the total nuclear intensity. The positions of the 2 N and 4 N DNA contents as well G0/G1, G2/M, and S phases are indicated ( n = 600 from 3 experiments). * P
    Figure Legend Snippet: Phenylephrine modulates cell size independently of cell cycle. (A) Representative immunofluorescence image showing hESC-CM stained positive for the myosin heavy chain α/β (MHCα/β, green), DAPI (blue), Ki67 (nuclear, red), and atrial natriuretic factor (ANF, perinuclear, orange) in the presence of cell cycle inhibitor blebbistatin (10 μM) at 30 days after differentiation. Scale bar represents 50 μm. Bar graphs showing cell size of hESC-CM (B), percentage of binucleated hESC-CM (C), and percentage of Ki67-positive hESC-CM (D) treated with phenylephrine (PE) in the presence of blebbistatin (Bleb, solid bar) or vehicle (light grey bar). Results are shown as mean ± SEM ( n > 100 MHC-positive cells per well, in triplicate, n = 2 preparations). The Cellomics Cell Cycle BioApplication classified hESC-CM treated with blebbistatin, nocodazole, or control medium into their cell cycle phase based on the total nuclear intensity of DNA binding DAPI (E). Cell cycle distribution presented as histogram where the Y -axis represents the number of instances and the X -axis represents the total nuclear intensity. The positions of the 2 N and 4 N DNA contents as well G0/G1, G2/M, and S phases are indicated ( n = 600 from 3 experiments). * P

    Techniques Used: Immunofluorescence, Staining, Binding Assay

    36) Product Images from "Modular cell-internalizing aptamer nanostructure enables targeted delivery of large functional RNAs in cancer cell lines"

    Article Title: Modular cell-internalizing aptamer nanostructure enables targeted delivery of large functional RNAs in cancer cell lines

    Journal: Nature Communications

    doi: 10.1038/s41467-018-04691-x

    Aptamer-mediated targeted delivery of large and small RNAs. Cell-targeting aptamers, Waz and C10.36, and their respective controls were assembled with 3WJdB ( a ) or AF488-anti-tail ( b ), and their ability to bind the same batch of NALM6 cells was assessed after 1 h incubation by flow cytometry. Similarly, Waz and C10.36, and their respective controls were annealed with AF488-labeled 3WJdB ( c ) or AF488-anti-tail ( d ), and their targeting properties were assessed using the same batch of NALM6 cells. Representative flow cytometry curves illustrate a shift in fluorescence for aptamer nanostructures bearing both Waz (blue) and C10.36 (magenta) assembled with the RNA payloads. Cells treated only with DFHBI-1T (20 μM) are shown in green. All non-targeted controls are shown in orange: free 3WJdB in a , control aptamer assembled with AF488-anti-tail in b , d , free 3WJdB-AF488 in c . Gray filled curves: unstained cells. Normalized cell counts are reported on the y- axis, while on the x -axis is shown a log scale of fluorescence intensity. Geometric mean fluorescence intensity of Waz (blue), C10.36 (magenta), and non-targeted controls (orange) are shown above the respective curves in c , d . All curves in c , d are representative of two independent experiments. Geometric mean fluorescence intensity values for leukemia cell lines are reported in e using 3WJdB stained with DFHBI-1T as payload and in f . Values are the mean ± SD for at least three independent experiments. Statistical analysis for comparing multiple groups in each cell line was analyzed by one-way ANOVA with post hoc Tukey’s test. Brackets with asterisks represent statistical difference: ns not significant; * p
    Figure Legend Snippet: Aptamer-mediated targeted delivery of large and small RNAs. Cell-targeting aptamers, Waz and C10.36, and their respective controls were assembled with 3WJdB ( a ) or AF488-anti-tail ( b ), and their ability to bind the same batch of NALM6 cells was assessed after 1 h incubation by flow cytometry. Similarly, Waz and C10.36, and their respective controls were annealed with AF488-labeled 3WJdB ( c ) or AF488-anti-tail ( d ), and their targeting properties were assessed using the same batch of NALM6 cells. Representative flow cytometry curves illustrate a shift in fluorescence for aptamer nanostructures bearing both Waz (blue) and C10.36 (magenta) assembled with the RNA payloads. Cells treated only with DFHBI-1T (20 μM) are shown in green. All non-targeted controls are shown in orange: free 3WJdB in a , control aptamer assembled with AF488-anti-tail in b , d , free 3WJdB-AF488 in c . Gray filled curves: unstained cells. Normalized cell counts are reported on the y- axis, while on the x -axis is shown a log scale of fluorescence intensity. Geometric mean fluorescence intensity of Waz (blue), C10.36 (magenta), and non-targeted controls (orange) are shown above the respective curves in c , d . All curves in c , d are representative of two independent experiments. Geometric mean fluorescence intensity values for leukemia cell lines are reported in e using 3WJdB stained with DFHBI-1T as payload and in f . Values are the mean ± SD for at least three independent experiments. Statistical analysis for comparing multiple groups in each cell line was analyzed by one-way ANOVA with post hoc Tukey’s test. Brackets with asterisks represent statistical difference: ns not significant; * p

    Techniques Used: Incubation, Flow Cytometry, Cytometry, Labeling, Fluorescence, Staining

    37) Product Images from "CRISPR/Cas9-mediated hypoxia inducible factor-1α knockout enhances the antitumor effect of transarterial embolization in hepatocellular carcinoma"

    Article Title: CRISPR/Cas9-mediated hypoxia inducible factor-1α knockout enhances the antitumor effect of transarterial embolization in hepatocellular carcinoma

    Journal: Oncology Reports

    doi: 10.3892/or.2018.6667

    Generation of the CRISPR/Cas9-based lentivirus and HIF-1 α knockout in SMMC-7721 cells and in the SMMC-7721-induced tumor tissues of mice. (A) Immunohistochemical analysis for the detection of HIF-1α expression in hepatocellular carcinoma tissues following infection with LV-Ctrl or LV-H721 in the subcutaneous animal model (bars, 100 µm). (B) Diagram illustration of the lentiviral vector ( Lenti-CAS9-sgRNA-egfp ). Three sgRNAs (sgRNA719, sgRNA720 and sgRNA721) targeting HIF-1α were designed, and Lenti-CAS9-sgRNA719 / 720 /721 plasmids were constructed. (C) Gel electrophoresis and DNA analysis of the HIF-1α genomic frame shift mutation conducted after the T7E1 endonuclease assay in SMMC-7721 cells infected with LV-Ctrl or LV-H719/720/721. (D) Western blot analysis of HIF-1α expression in the different lentivirus-infected SMMC-7721 cells with CoCl 2 -simulated hypoxia. (E) GFP-positive cells analyzed by flow cytometry following infection with LV-Ctrl and LV-H721. (F) Relative mRNA and (G) protein expression levels of VEGF and MDR1 in different cell groups were examined by reverse transcription-quantitative polymerase chain reaction and western blot analysis. β-actin served as an internal control. Data are a representation of four repeated experiments, and histograms represent the mean ± standard deviation. ***P
    Figure Legend Snippet: Generation of the CRISPR/Cas9-based lentivirus and HIF-1 α knockout in SMMC-7721 cells and in the SMMC-7721-induced tumor tissues of mice. (A) Immunohistochemical analysis for the detection of HIF-1α expression in hepatocellular carcinoma tissues following infection with LV-Ctrl or LV-H721 in the subcutaneous animal model (bars, 100 µm). (B) Diagram illustration of the lentiviral vector ( Lenti-CAS9-sgRNA-egfp ). Three sgRNAs (sgRNA719, sgRNA720 and sgRNA721) targeting HIF-1α were designed, and Lenti-CAS9-sgRNA719 / 720 /721 plasmids were constructed. (C) Gel electrophoresis and DNA analysis of the HIF-1α genomic frame shift mutation conducted after the T7E1 endonuclease assay in SMMC-7721 cells infected with LV-Ctrl or LV-H719/720/721. (D) Western blot analysis of HIF-1α expression in the different lentivirus-infected SMMC-7721 cells with CoCl 2 -simulated hypoxia. (E) GFP-positive cells analyzed by flow cytometry following infection with LV-Ctrl and LV-H721. (F) Relative mRNA and (G) protein expression levels of VEGF and MDR1 in different cell groups were examined by reverse transcription-quantitative polymerase chain reaction and western blot analysis. β-actin served as an internal control. Data are a representation of four repeated experiments, and histograms represent the mean ± standard deviation. ***P

    Techniques Used: CRISPR, Knock-Out, Mouse Assay, Immunohistochemistry, Expressing, Infection, Animal Model, Plasmid Preparation, Construct, Nucleic Acid Electrophoresis, Mutagenesis, Western Blot, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction, Standard Deviation

    38) Product Images from "Mice harboring the human SLC30A8 R138X loss-of-function mutation have increased insulin secretory capacity"

    Article Title: Mice harboring the human SLC30A8 R138X loss-of-function mutation have increased insulin secretory capacity

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1721418115

    Analysis of Slc30a8 RNA and protein in islets from male R138X mice on chow diet. ( A ) Slc30a8 RNA in situ hybridization of pancreatic islets isolated from wild-type, knockout, and R138X mice. KO islets were used as negative control. Red, glucagon RNA; green, insulin RNA; white, Slc30a8 RNA. ( B ) Quantification of islet Slc30a8 RNA levels using qPCR analysis. n.d., not detected. ( C ) Western blot of islets isolated from chow-fed WT, KO, and R138X mice. KO islets were used as negative control. The arrow indicates SLC30A8 protein; asterisks denote unspecific bands. ( D ) Dithizone staining of pancreatic islets isolated from WT, KO, and R138X mice.
    Figure Legend Snippet: Analysis of Slc30a8 RNA and protein in islets from male R138X mice on chow diet. ( A ) Slc30a8 RNA in situ hybridization of pancreatic islets isolated from wild-type, knockout, and R138X mice. KO islets were used as negative control. Red, glucagon RNA; green, insulin RNA; white, Slc30a8 RNA. ( B ) Quantification of islet Slc30a8 RNA levels using qPCR analysis. n.d., not detected. ( C ) Western blot of islets isolated from chow-fed WT, KO, and R138X mice. KO islets were used as negative control. The arrow indicates SLC30A8 protein; asterisks denote unspecific bands. ( D ) Dithizone staining of pancreatic islets isolated from WT, KO, and R138X mice.

    Techniques Used: Mouse Assay, RNA In Situ Hybridization, Isolation, Knock-Out, Negative Control, Real-time Polymerase Chain Reaction, Western Blot, Staining

    39) Product Images from "Pospiviroid Infection of Tomato Regulates the Expression of Genes Involved in Flower and Fruit Development"

    Article Title: Pospiviroid Infection of Tomato Regulates the Expression of Genes Involved in Flower and Fruit Development

    Journal: Viruses

    doi: 10.3390/v10100516

    Stem-loop quantitative RT-PCR of miRNA accumulation in TPMVd-infected leaf tissue 4 w. p.i. relative to mock-inoculated controls to which a value of 1 was assigned. U6snRNA was used as an internal reference. Error bars indicate two times the value of SD for the corresponding data set. All fold changes were statistically significant from mock-inoculated controls when analyzed using a paired t -test ( p
    Figure Legend Snippet: Stem-loop quantitative RT-PCR of miRNA accumulation in TPMVd-infected leaf tissue 4 w. p.i. relative to mock-inoculated controls to which a value of 1 was assigned. U6snRNA was used as an internal reference. Error bars indicate two times the value of SD for the corresponding data set. All fold changes were statistically significant from mock-inoculated controls when analyzed using a paired t -test ( p

    Techniques Used: Quantitative RT-PCR, Infection

    40) Product Images from "Stemness and differentiation potential-recovery effects of sinapic acid against ultraviolet-A-induced damage through the regulation of p38 MAPK and NF-κB"

    Article Title: Stemness and differentiation potential-recovery effects of sinapic acid against ultraviolet-A-induced damage through the regulation of p38 MAPK and NF-κB

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-01089-5

    Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF gene expression. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) Intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. ( B ) The triglyceride content was measured using a triglyceride assay kit (Cayman Chemical, Ann Arbor, MI). The results were verified by three repetitions of the experiments, each of which was conducted in triplicate. ( C ) At day 14 after the induction of differentiation, the supernatants were harvested for MIF measurement. Data are expressed as the means ± S.D. ( D ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the MIF gene were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. * p
    Figure Legend Snippet: Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF gene expression. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) Intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. ( B ) The triglyceride content was measured using a triglyceride assay kit (Cayman Chemical, Ann Arbor, MI). The results were verified by three repetitions of the experiments, each of which was conducted in triplicate. ( C ) At day 14 after the induction of differentiation, the supernatants were harvested for MIF measurement. Data are expressed as the means ± S.D. ( D ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the MIF gene were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. * p

    Techniques Used: Expressing, Irradiation, Staining, Isolation, Quantitative RT-PCR

    Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF-AMPK-KLF2 signaling by inhibiting NF-κB. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) At 14 days after the induction of differentiation, the total lysates were analyzed by Western blot using the indicated antibodies. ( B ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the KLF2 and PPARγ genes were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. ( C ) At 1 h after the induction of differentiation, cell lysates were analyzed using a Multi-Target Sandwich ELISA Kit. All of the results were verified by repeating the experiments, each of which was conducted in duplicate, three times. SA: sinapic acid, DIF: differentiation media, UVA: ultraviolet A. Data are expressed as the means ± S.D. * p
    Figure Legend Snippet: Sinapic acid attenuates UVA-induced suppression of adipogenic differentiation through downregulation of MIF-AMPK-KLF2 signaling by inhibiting NF-κB. Two-day post confluent hAMSCs (day 0) were irradiated with UVA (5 J/cm 2 ) and then treated with sinapic acid (400 μM), followed by stimulation with STEM PRO ® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO ® adipocyte differentiation media every three days until the end of the experiment at day 14. These assays were performed on fully differentiated adipocytes (day 14). ( A ) At 14 days after the induction of differentiation, the total lysates were analyzed by Western blot using the indicated antibodies. ( B ) At 14 days after the induction of differentiation, the total RNA was isolated and the mRNA levels of the KLF2 and PPARγ genes were measured by real-time quantitative RT-PCR. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. ( C ) At 1 h after the induction of differentiation, cell lysates were analyzed using a Multi-Target Sandwich ELISA Kit. All of the results were verified by repeating the experiments, each of which was conducted in duplicate, three times. SA: sinapic acid, DIF: differentiation media, UVA: ultraviolet A. Data are expressed as the means ± S.D. * p

    Techniques Used: Irradiation, Western Blot, Isolation, Quantitative RT-PCR, Sandwich ELISA

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