qiasymphony system  (Qiagen)

 
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    Name:
    QIAsymphony AS
    Description:
    For fully integrated automation of complete workflows from sample preparation to assay setup Kit contents QIAsymphony assay setup module includes1 year warranty on parts and labor Benefits Innovative easy to use modular system with built in touchscreen Purification of DNA and RNA from a wide range of samples Automatic transfer of eluates to the QIAsymphony AS for assay setup Continuous sample loading with bar code reading for sample tracking Import of sample lists and export of sampl
    Catalog Number:
    9001301
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    Category:
    QIAsymphony SP AS instruments
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    Structured Review

    Qiagen qiasymphony system
    QIAsymphony AS
    For fully integrated automation of complete workflows from sample preparation to assay setup Kit contents QIAsymphony assay setup module includes1 year warranty on parts and labor Benefits Innovative easy to use modular system with built in touchscreen Purification of DNA and RNA from a wide range of samples Automatic transfer of eluates to the QIAsymphony AS for assay setup Continuous sample loading with bar code reading for sample tracking Import of sample lists and export of sampl
    https://www.bioz.com/result/qiasymphony system/product/Qiagen
    Average 99 stars, based on 7230 article reviews
    Price from $9.99 to $1999.99
    qiasymphony system - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.
    Figure Legend Snippet: Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.

    Techniques Used: Standard Deviation

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    2) Product Images from "Inhibition of MMP-9 expression by ritonavir or saquinavir is associated with inactivation of the AKT/Fra-1 pathway in cervical intraepithelial neoplasia cells"

    Article Title: Inhibition of MMP-9 expression by ritonavir or saquinavir is associated with inactivation of the AKT/Fra-1 pathway in cervical intraepithelial neoplasia cells

    Journal: Oncology Letters

    doi: 10.3892/ol.2017.5835

    SQV and RTV counteract EGF-induced cell invasion, MMP-9 expression, AKT phosphorylation and nuclear Fra-1 protein expression in W12 cells. W12 cells were cultured for 96 h in the presence of 10 µM SQV or RTV, or in their absence (control). (A) Cells were stimulated to invade a reconstituted basement membrane in response to 20 ng/ml human recombinant EGF, or to its suspension buffer (0.1% bovine serum albumin in phosphate-buffered saline, indicated here as EGF 0 ng/ml). Results are expressed as the mean ± standard deviation from 3 experiments, each performed in duplicate chambers. (B) Representative zymography of EGF-supplemented, serum-free supernatants. The de-stained areas indicate gelatinolytic activity corresponding to MMP-9 (92 kDa) released by the cells. (C) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-9 messenger RNA levels (relative to GADPH) in cells cultured in EGF-supplemented growth medium, in the absence or presence of 10 µM SQV/RTV. Results are expressed as the mean ± standard deviation from 3 experiments. (D) Representative western blot analysis and quantification by densitometry of pAKT protein levels (relative to β-actin) in W12 cells lysed following a 30-min exposure to EGF. (E) Representative western blot analysis and quantification by densitometry of nuclear Fra-1 protein levels (relative to C23) in W12 cells lysed following a 6-h exposure to EGF. *P
    Figure Legend Snippet: SQV and RTV counteract EGF-induced cell invasion, MMP-9 expression, AKT phosphorylation and nuclear Fra-1 protein expression in W12 cells. W12 cells were cultured for 96 h in the presence of 10 µM SQV or RTV, or in their absence (control). (A) Cells were stimulated to invade a reconstituted basement membrane in response to 20 ng/ml human recombinant EGF, or to its suspension buffer (0.1% bovine serum albumin in phosphate-buffered saline, indicated here as EGF 0 ng/ml). Results are expressed as the mean ± standard deviation from 3 experiments, each performed in duplicate chambers. (B) Representative zymography of EGF-supplemented, serum-free supernatants. The de-stained areas indicate gelatinolytic activity corresponding to MMP-9 (92 kDa) released by the cells. (C) Reverse transcription-quantitative polymerase chain reaction analysis of MMP-9 messenger RNA levels (relative to GADPH) in cells cultured in EGF-supplemented growth medium, in the absence or presence of 10 µM SQV/RTV. Results are expressed as the mean ± standard deviation from 3 experiments. (D) Representative western blot analysis and quantification by densitometry of pAKT protein levels (relative to β-actin) in W12 cells lysed following a 30-min exposure to EGF. (E) Representative western blot analysis and quantification by densitometry of nuclear Fra-1 protein levels (relative to C23) in W12 cells lysed following a 6-h exposure to EGF. *P

    Techniques Used: Expressing, Cell Culture, Recombinant, Standard Deviation, Zymography, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot

    3) Product Images from "Keeping it cool: Soil sample cold pack storage and DNA shipment up to 1 month does not impact metabarcoding results. Soil sample cold pack storage and DNA shipment up to 1 month does not impact metabarcoding results"

    Article Title: Keeping it cool: Soil sample cold pack storage and DNA shipment up to 1 month does not impact metabarcoding results. Soil sample cold pack storage and DNA shipment up to 1 month does not impact metabarcoding results

    Journal: Ecology and Evolution

    doi: 10.1002/ece3.6219

    Gene copy number for 16S (bacteria; a) and ITS (fungi; b) based on storage and location, with 95% confidence intervals. Lower gene copies of both bacteria (a, p = 1.62E−08) and fungi (b, p = 2.55E−04) were found in Saskatchewan for samples stored in RNAlater
    Figure Legend Snippet: Gene copy number for 16S (bacteria; a) and ITS (fungi; b) based on storage and location, with 95% confidence intervals. Lower gene copies of both bacteria (a, p = 1.62E−08) and fungi (b, p = 2.55E−04) were found in Saskatchewan for samples stored in RNAlater

    Techniques Used:

    4) Product Images from "RNA Helicase DDX1 Converts RNA G-Quadruplex Structures into R-Loops to Promote IgH Class Switch Recombination"

    Article Title: RNA Helicase DDX1 Converts RNA G-Quadruplex Structures into R-Loops to Promote IgH Class Switch Recombination

    Journal: Molecular Cell

    doi: 10.1016/j.molcel.2018.04.001

    Impaired CSR in DDX1 Knockout B Cells (A) Diagram of conditional DDX1 ( DDX1 loxP ) and Cre-deleted DDX1 ( DDX1 del ) alleles. PCR primers are indicated. (B) PCR analysis of genomic DNA from DDX1 loxP/loxP splenic B cells treated with TAT-Cre (2 replicates). (C) Western blot of WT and DDX1-deleted (DDX1Δ) B cells (day 4, 2 replicates). (D) Flow cytometric analysis for surface IgG3 and IgG1 expression and CFSE in WT and DDX1Δ B cells stimulated for 4 days. (E) Quantification of CSR for cultures shown in (D). Each symbol represents B cell cultures from individual mice (n ≥ 3, mean ± SD). (F) Percentage of IgG1 + cells analyzed per number of cell divisions in CFSE-labeled LPS plus IL-4 cultures at day 4. One representative experiment with 3 mice per genotype (mean ± SD). (G) Serum Ig concentrations in naive WT and DDX1 cKO mice. Each symbol represents individual mice and horizontal lines indicate the mean. (H) Antigen-specific IgG and IgM immune responses in WT and DDX1 cKO mice immunized with ovalbumin (OVA) antigen. Serum was analyzed by ELISA and endpoint titers were determined (n = 2, mean ± SD; 3–7 mice in each time point except week 10 where 2–3 mice were analyzed). See also Figure S1 .
    Figure Legend Snippet: Impaired CSR in DDX1 Knockout B Cells (A) Diagram of conditional DDX1 ( DDX1 loxP ) and Cre-deleted DDX1 ( DDX1 del ) alleles. PCR primers are indicated. (B) PCR analysis of genomic DNA from DDX1 loxP/loxP splenic B cells treated with TAT-Cre (2 replicates). (C) Western blot of WT and DDX1-deleted (DDX1Δ) B cells (day 4, 2 replicates). (D) Flow cytometric analysis for surface IgG3 and IgG1 expression and CFSE in WT and DDX1Δ B cells stimulated for 4 days. (E) Quantification of CSR for cultures shown in (D). Each symbol represents B cell cultures from individual mice (n ≥ 3, mean ± SD). (F) Percentage of IgG1 + cells analyzed per number of cell divisions in CFSE-labeled LPS plus IL-4 cultures at day 4. One representative experiment with 3 mice per genotype (mean ± SD). (G) Serum Ig concentrations in naive WT and DDX1 cKO mice. Each symbol represents individual mice and horizontal lines indicate the mean. (H) Antigen-specific IgG and IgM immune responses in WT and DDX1 cKO mice immunized with ovalbumin (OVA) antigen. Serum was analyzed by ELISA and endpoint titers were determined (n = 2, mean ± SD; 3–7 mice in each time point except week 10 where 2–3 mice were analyzed). See also Figure S1 .

    Techniques Used: Knock-Out, Polymerase Chain Reaction, Western Blot, Flow Cytometry, Expressing, Mouse Assay, Labeling, Enzyme-linked Immunosorbent Assay

    5) Product Images from "Multiple Targets of Nitric Oxide in the Tricarboxylic Acid (TCA) Cycle of Salmonella enterica Serovar Typhimurium"

    Article Title: Multiple Targets of Nitric Oxide in the Tricarboxylic Acid (TCA) Cycle of Salmonella enterica Serovar Typhimurium

    Journal: Cell host & microbe

    doi: 10.1016/j.chom.2011.06.004

    NO· creates a regulatory block that inhibits the reductive branch of the TCA cycle A. Expression levels of sdhC or frdA as determined by Q-RT PCR of three independent RNA samples from WT and isogenic Δ fnr S. Typhimurium strain 14028s either untreated (blue columns) or exposed to NO· (red columns). Transcript levels relative to rpoD were determined by a modified ΔΔC t method as previously described. B. Expressing the reductive TCA cycle in the presence of NO· restores MK prototrophy to S. Typhimurium. The frdABCD genes from 14028s were cloned and expressed from an IPTG-inducible promoter (p trc ). IPTG was added at 100 μM for each growth curve. M9 Glc medium was either unsupplemented (None), supplemented with all 20 amino acids (All), or with a mixture of amino acids lacking both Met and Lys (−MK). pTrc99a was used as a vector control. A Δ sdh mutation was introduced to prevent futile cycling by high expression of both SDH and FRD activities.
    Figure Legend Snippet: NO· creates a regulatory block that inhibits the reductive branch of the TCA cycle A. Expression levels of sdhC or frdA as determined by Q-RT PCR of three independent RNA samples from WT and isogenic Δ fnr S. Typhimurium strain 14028s either untreated (blue columns) or exposed to NO· (red columns). Transcript levels relative to rpoD were determined by a modified ΔΔC t method as previously described. B. Expressing the reductive TCA cycle in the presence of NO· restores MK prototrophy to S. Typhimurium. The frdABCD genes from 14028s were cloned and expressed from an IPTG-inducible promoter (p trc ). IPTG was added at 100 μM for each growth curve. M9 Glc medium was either unsupplemented (None), supplemented with all 20 amino acids (All), or with a mixture of amino acids lacking both Met and Lys (−MK). pTrc99a was used as a vector control. A Δ sdh mutation was introduced to prevent futile cycling by high expression of both SDH and FRD activities.

    Techniques Used: Blocking Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Modification, Clone Assay, Gas Chromatography, Plasmid Preparation, Mutagenesis

    6) Product Images from "Genetic variants of calcium and vitamin D metabolism in kidney stone disease"

    Article Title: Genetic variants of calcium and vitamin D metabolism in kidney stone disease

    Journal: Nature Communications

    doi: 10.1038/s41467-019-13145-x

    CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p
    Figure Legend Snippet: CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Concentration Assay, Incubation

    7) Product Images from "Abdominal microbial communities in ants depend on colony membership rather than caste and are linked to colony productivity. Abdominal microbial communities in ants depend on colony membership rather than caste and are linked to colony productivity"

    Article Title: Abdominal microbial communities in ants depend on colony membership rather than caste and are linked to colony productivity. Abdominal microbial communities in ants depend on colony membership rather than caste and are linked to colony productivity

    Journal: Ecology and Evolution

    doi: 10.1002/ece3.5801

    An overview of the sampling procedure and subsequent high‐throughput sequencing of the 16S rRNA gene. In May, fourteen Temnothorax nylanderi colonies were collected from the forest and kept in the laboratory for 2 months. Although 14 colonies were initially collected from the forest in May, from only 11 colonies we have worker samples for the Field May and Laboratory July sampling group, due to loss of samples from three colonies. In July, 10 new colonies were collected from the same field site
    Figure Legend Snippet: An overview of the sampling procedure and subsequent high‐throughput sequencing of the 16S rRNA gene. In May, fourteen Temnothorax nylanderi colonies were collected from the forest and kept in the laboratory for 2 months. Although 14 colonies were initially collected from the forest in May, from only 11 colonies we have worker samples for the Field May and Laboratory July sampling group, due to loss of samples from three colonies. In July, 10 new colonies were collected from the same field site

    Techniques Used: Sampling, Next-Generation Sequencing

    8) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.
    Figure Legend Snippet: Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.

    Techniques Used: Standard Deviation

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    9) Product Images from "Deregulation of CREB Signaling Pathway Induced by Chronic Hyperglycemia Downregulates NeuroD Transcription"

    Article Title: Deregulation of CREB Signaling Pathway Induced by Chronic Hyperglycemia Downregulates NeuroD Transcription

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0034860

    Chronic hyperglycemia alters the responsiveness to cAMP in pancreatic islets. ( A ) Rat islet cells were cultured in the presence of 5 mM or 30 mM glucose for 8 days and stimulated with 30 µM forskolin for 0–12 h after 2 h preconditioning in 5 mM glucose. ( B∼F ) Real-time PCR was carried out using SYBR green to quantitate the mRNA levels of indicated genes in diverse conditions shown in A . Relative mRNA levels were estimated from Ct values summarized in Supporting Table S1 using 2 −ΔΔCt method. The data from three independent experiments are presented as average fold ratios of relative mRNA expression compare with the 5 mM glucose-cultured islets before forskolin treatment in 5 mM glucose-cultured islets. Significant effects of forskolin (*, P
    Figure Legend Snippet: Chronic hyperglycemia alters the responsiveness to cAMP in pancreatic islets. ( A ) Rat islet cells were cultured in the presence of 5 mM or 30 mM glucose for 8 days and stimulated with 30 µM forskolin for 0–12 h after 2 h preconditioning in 5 mM glucose. ( B∼F ) Real-time PCR was carried out using SYBR green to quantitate the mRNA levels of indicated genes in diverse conditions shown in A . Relative mRNA levels were estimated from Ct values summarized in Supporting Table S1 using 2 −ΔΔCt method. The data from three independent experiments are presented as average fold ratios of relative mRNA expression compare with the 5 mM glucose-cultured islets before forskolin treatment in 5 mM glucose-cultured islets. Significant effects of forskolin (*, P

    Techniques Used: Cell Culture, Real-time Polymerase Chain Reaction, SYBR Green Assay, Expressing

    Chronic hyperglycemia alters the β-cell specific gene expression in pancreatic islets. ( A ) Rat islet cells were cultured in the presence of 5 mM or 30 mM glucose for 8 days, and stimulated with 15 mM glucose for 0–6 h after 2 h preconditioning in 5 mM glucose. ( B ) Dithizone staining of islet cells after 8 days of culture. Scale bar: 100 µm. ( C ) In low (5 mM) glucose condition, insulin secretion was normal in response to acute stimulation with 15 mM glucose after 8 day culture whereas insulin secretion was impaired after 8-day exposure to high (30 mM) glucose. Under high glucose conditions, cellular insulin content were significantly reduced, and total proteins were decreased slightly. ( D∼H ) Real-time PCR was carried out using SYBR green to quantitate the mRNA levels of indicated genes in diverse conditions shown in A . Relative mRNA levels were estimated from of Ct values summarized in Supporting Table S1 using 2 −ΔΔCt method. The data from three independent experiments are presented as average fold ratios (means ± S.E.) of relative mRNA expression compare with the 5 mM glucose-cultured islets before glucose stimulation. Simultaneous decreases in the mRNA levels and the intracellular insulin content correlated well. Significant effects of 15 mM glucose (*, P
    Figure Legend Snippet: Chronic hyperglycemia alters the β-cell specific gene expression in pancreatic islets. ( A ) Rat islet cells were cultured in the presence of 5 mM or 30 mM glucose for 8 days, and stimulated with 15 mM glucose for 0–6 h after 2 h preconditioning in 5 mM glucose. ( B ) Dithizone staining of islet cells after 8 days of culture. Scale bar: 100 µm. ( C ) In low (5 mM) glucose condition, insulin secretion was normal in response to acute stimulation with 15 mM glucose after 8 day culture whereas insulin secretion was impaired after 8-day exposure to high (30 mM) glucose. Under high glucose conditions, cellular insulin content were significantly reduced, and total proteins were decreased slightly. ( D∼H ) Real-time PCR was carried out using SYBR green to quantitate the mRNA levels of indicated genes in diverse conditions shown in A . Relative mRNA levels were estimated from of Ct values summarized in Supporting Table S1 using 2 −ΔΔCt method. The data from three independent experiments are presented as average fold ratios (means ± S.E.) of relative mRNA expression compare with the 5 mM glucose-cultured islets before glucose stimulation. Simultaneous decreases in the mRNA levels and the intracellular insulin content correlated well. Significant effects of 15 mM glucose (*, P

    Techniques Used: Expressing, Cell Culture, Staining, Real-time Polymerase Chain Reaction, SYBR Green Assay

    10) Product Images from "FOXP3 expression is modulated by TGF-β1/NOTCH1 pathway in human melanoma"

    Article Title: FOXP3 expression is modulated by TGF-β1/NOTCH1 pathway in human melanoma

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2018.3618

    FOXP3 expression in human melanoma cell lines at different stages. (A) RT-qPCR of FOXP3 in melanocytes (NHEM), primary (WM35) and metastatic (A375 and A2058) melanoma cells. The melanoma cell lines expressed FOXP3 mRNA. Melanocytes served as a control. A375 and A2058 cells showed the highest levels of FOXP3 gene expression. (B and C) Effect of TGFβ-1 treatment on protein and FOXP3 mRNA levels in melanoma cell lines. Treatment with rhTGF-β1 (5 ng/ml) for 48 h induced a higher increase of FOXP3 mRNA and their own protein levels in WM35, A375 and A2058 melanoma cells. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of multiple groups was performed by ANOVA and Tukey's test. High significance ( *** P
    Figure Legend Snippet: FOXP3 expression in human melanoma cell lines at different stages. (A) RT-qPCR of FOXP3 in melanocytes (NHEM), primary (WM35) and metastatic (A375 and A2058) melanoma cells. The melanoma cell lines expressed FOXP3 mRNA. Melanocytes served as a control. A375 and A2058 cells showed the highest levels of FOXP3 gene expression. (B and C) Effect of TGFβ-1 treatment on protein and FOXP3 mRNA levels in melanoma cell lines. Treatment with rhTGF-β1 (5 ng/ml) for 48 h induced a higher increase of FOXP3 mRNA and their own protein levels in WM35, A375 and A2058 melanoma cells. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of multiple groups was performed by ANOVA and Tukey's test. High significance ( *** P

    Techniques Used: Expressing, Quantitative RT-PCR

    Effect of GSI on FOXP3 expression in melanoma cell lines. (A) Inhibition of FOXP3 mRNA is shown after 72 h of 5-, 10- and 20 μ M GSI treatment in melanoma cells. RT-qPCR shows that FOXP3 mRNA levels were downregulated in GSI-treated WM35, A375 and A2058 melanoma cells in a dose-dependent manner. Maximum inhibition of FOXP3 was observed at 20 μ M of GSI. (B) Inhibition of FOXP3 mRNA after 24, 48 and 72 h with 20 μ M/GSI treatment in WM35, A375 and A2058 melanoma cell lines. A statistically significant time-dependent decrease in FOXP3 mRNA level was observed in each melanoma cell line. (C) Inhibition of FOXP3 protein expression after 72 h of 5-, 10- and 20 μ M GSI treatment in WM35 melanoma cells. Western blot analysis showed that the protein levels of FOXP3 were downregulated in GSI-treated WM35 cells in a dose-dependent manner. GAPDH expression was used as a loading control. (D and E) Effect of GSI/TGF-β1 treatment on FOXP3 mRNA and protein expression in melanoma cell lines. Inhibition of FOXP3 mRNA and protein levels are shown after 72 h of GSI treatment in WM35, A375 and A2058 melanoma cells. In vitro GSI treatment downregulated TGF-β1-induced FOXP3 mRNA and protein levels in all the melanoma cell lines. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of mRNA FOXP3 expression in multiple groups was performed by ANOVA and Tukey's test. GSI, γ-secretase inhibitor; FOXP3, forkhead box protein 3; TGF-β, transforming growth factor-β; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. * P
    Figure Legend Snippet: Effect of GSI on FOXP3 expression in melanoma cell lines. (A) Inhibition of FOXP3 mRNA is shown after 72 h of 5-, 10- and 20 μ M GSI treatment in melanoma cells. RT-qPCR shows that FOXP3 mRNA levels were downregulated in GSI-treated WM35, A375 and A2058 melanoma cells in a dose-dependent manner. Maximum inhibition of FOXP3 was observed at 20 μ M of GSI. (B) Inhibition of FOXP3 mRNA after 24, 48 and 72 h with 20 μ M/GSI treatment in WM35, A375 and A2058 melanoma cell lines. A statistically significant time-dependent decrease in FOXP3 mRNA level was observed in each melanoma cell line. (C) Inhibition of FOXP3 protein expression after 72 h of 5-, 10- and 20 μ M GSI treatment in WM35 melanoma cells. Western blot analysis showed that the protein levels of FOXP3 were downregulated in GSI-treated WM35 cells in a dose-dependent manner. GAPDH expression was used as a loading control. (D and E) Effect of GSI/TGF-β1 treatment on FOXP3 mRNA and protein expression in melanoma cell lines. Inhibition of FOXP3 mRNA and protein levels are shown after 72 h of GSI treatment in WM35, A375 and A2058 melanoma cells. In vitro GSI treatment downregulated TGF-β1-induced FOXP3 mRNA and protein levels in all the melanoma cell lines. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of mRNA FOXP3 expression in multiple groups was performed by ANOVA and Tukey's test. GSI, γ-secretase inhibitor; FOXP3, forkhead box protein 3; TGF-β, transforming growth factor-β; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. * P

    Techniques Used: Expressing, Inhibition, Quantitative RT-PCR, Western Blot, In Vitro

    Expression of NOTCH1 NICD and NOTCH-specific target gene HES1 in melanoma cell lines. (A) mRNA of NOTCH1 NICD and HES1 was measured by RT-qPCR in NHEM, WM35 and A375 and A2058 melanoma cells. Melanocytes served as the control. WM35 showed a higher level of NOTCH1 NICD mRNA and HES1 mRNA than A375 and A2058 cells. (B) Protein level of NOTCH1 NICD and HES1 was measured by western blot analysis in WM35, A375 and A2058 melanoma cell lines. All of the melanoma cell lines positively expressed NOTCH1 NICD and HES1. (B-D) Effect of TGFβ-1 treatment on NOTCH1 NICD , HES1 mRNA and protein levels in melanoma cell lines. Treatment with rhTGF-β1 (5 ng/ml) for 48 h induced a higher increase of NOTCH1 NICD and HES1 mRNA and their own protein levels in WM35, A375 and A2058 melanoma cells. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of mRNA NOTCH-1 and HES1 expression in multiple groups was performed by ANOVA and Tukey's test. HES1, hairy and enhancer of split 1; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor-β; rh, recombinant human; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. ** P
    Figure Legend Snippet: Expression of NOTCH1 NICD and NOTCH-specific target gene HES1 in melanoma cell lines. (A) mRNA of NOTCH1 NICD and HES1 was measured by RT-qPCR in NHEM, WM35 and A375 and A2058 melanoma cells. Melanocytes served as the control. WM35 showed a higher level of NOTCH1 NICD mRNA and HES1 mRNA than A375 and A2058 cells. (B) Protein level of NOTCH1 NICD and HES1 was measured by western blot analysis in WM35, A375 and A2058 melanoma cell lines. All of the melanoma cell lines positively expressed NOTCH1 NICD and HES1. (B-D) Effect of TGFβ-1 treatment on NOTCH1 NICD , HES1 mRNA and protein levels in melanoma cell lines. Treatment with rhTGF-β1 (5 ng/ml) for 48 h induced a higher increase of NOTCH1 NICD and HES1 mRNA and their own protein levels in WM35, A375 and A2058 melanoma cells. As an internal control, GAPDH was used for normalization. Data are shown as mean ± SD of three independent experiments. The comparison of mRNA NOTCH-1 and HES1 expression in multiple groups was performed by ANOVA and Tukey's test. HES1, hairy and enhancer of split 1; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; TGF-β, transforming growth factor-β; rh, recombinant human; GAPDH, glyceraldehyde 3-phosphate dehydrogenase. ** P

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

    11) Product Images from "Gastrointestinal cancer cells treatment with bevacizumab activates a VEGF autoregulatory mechanism involving telomerase catalytic subunit hTERT via PI3K-AKT, HIF-1α and VEGF receptors"

    Article Title: Gastrointestinal cancer cells treatment with bevacizumab activates a VEGF autoregulatory mechanism involving telomerase catalytic subunit hTERT via PI3K-AKT, HIF-1α and VEGF receptors

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0179202

    Bevacizumab increases VEGFR1 and VEGFR2 expression. (A, B) Telomerase inhibitors were combined to bevacizumab (100 μ g/ml) and then VEGFR1 (A) and VEGFR2 (B) were quantified by real-time PCR. (C, D) Cells were transiently transfected with either control siRNA or hTERT siRNA and treated with bevacizumab (100 μ g/ml). VEGFR1 (C) and VEGFR2 (D) were quantified by real-time qPCR. (E, F) HUVECs were cultured in the presence of telomerase inhibitors for 48 h and hTERT siRNA for 72 h. The expression of VEGFR1 (E) and VEGFR2 (F) were quantified by real-time PCR. Results were expressed as the mean ± SD from three experiments.
    Figure Legend Snippet: Bevacizumab increases VEGFR1 and VEGFR2 expression. (A, B) Telomerase inhibitors were combined to bevacizumab (100 μ g/ml) and then VEGFR1 (A) and VEGFR2 (B) were quantified by real-time PCR. (C, D) Cells were transiently transfected with either control siRNA or hTERT siRNA and treated with bevacizumab (100 μ g/ml). VEGFR1 (C) and VEGFR2 (D) were quantified by real-time qPCR. (E, F) HUVECs were cultured in the presence of telomerase inhibitors for 48 h and hTERT siRNA for 72 h. The expression of VEGFR1 (E) and VEGFR2 (F) were quantified by real-time PCR. Results were expressed as the mean ± SD from three experiments.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Cell Culture

    12) Product Images from "Gastrointestinal cancer cells treatment with bevacizumab activates a VEGF autoregulatory mechanism involving telomerase catalytic subunit hTERT via PI3K-AKT, HIF-1α and VEGF receptors"

    Article Title: Gastrointestinal cancer cells treatment with bevacizumab activates a VEGF autoregulatory mechanism involving telomerase catalytic subunit hTERT via PI3K-AKT, HIF-1α and VEGF receptors

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0179202

    Telomerase inhibition decreases the proliferation and tube formation of HUVECs. (A) HUVECs were treated with BIBR-1532 and costunolide. VEGFR2 protein levels were quantified by ELISA using the cell protein extracts. (B, C) HUVECs were transiently transfected with either a control siRNA or hTERT siRNA for 72 h. VEGFR1 and VEGFR2 expression was then assessed by real-time PCR. (D) HUVECs treated with BIBR-1532 and costunolide were collected and counted using trypan blue. (E) HUVECs were cultured in a 96-well plate and treated with telomerase inhibitors. Tetrazolium salt was added to each well and changes in absorbance following formazan formation was detected at 450 nm. (F, G) HUVECs were cultured on an ECM in the presence of recombinant VEGF 165 (50 ng/ml), telomerase inhibitors and bevacizumab. Capillary-like structures were quantified in the presence (F) and absence (G) of recombinant VEGF 165 .Values are represented as the mean ± SE of 10 randomly chosen fields.
    Figure Legend Snippet: Telomerase inhibition decreases the proliferation and tube formation of HUVECs. (A) HUVECs were treated with BIBR-1532 and costunolide. VEGFR2 protein levels were quantified by ELISA using the cell protein extracts. (B, C) HUVECs were transiently transfected with either a control siRNA or hTERT siRNA for 72 h. VEGFR1 and VEGFR2 expression was then assessed by real-time PCR. (D) HUVECs treated with BIBR-1532 and costunolide were collected and counted using trypan blue. (E) HUVECs were cultured in a 96-well plate and treated with telomerase inhibitors. Tetrazolium salt was added to each well and changes in absorbance following formazan formation was detected at 450 nm. (F, G) HUVECs were cultured on an ECM in the presence of recombinant VEGF 165 (50 ng/ml), telomerase inhibitors and bevacizumab. Capillary-like structures were quantified in the presence (F) and absence (G) of recombinant VEGF 165 .Values are represented as the mean ± SE of 10 randomly chosen fields.

    Techniques Used: Inhibition, Enzyme-linked Immunosorbent Assay, Transfection, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Recombinant

    Telomerase regulates VEGF secretion and expression independently of telomerase activity. (A, B) VEGF secretion was assessed by ELISA following a 48-h treatment with telomerase inhibitors BIBR-1532 (10 μ M) and costunolide (10 μ M), and after 72 h of hTERT knockdown with siRNA. (C, D) Total RNAs were isolated from the cells treated in (A), and VEGF expression was analyzed by real-time PCR with GAPDH as the internal control. (E, F, G) Saos-2 cells were transiently transfected with an empty vector, hTERT-WT, and hTERT-DN. hTERT (E) and VEGF expression (F) were quantified by real-time PCR. Secreted VEGF was quantified by ELISA (G). Results were expressed as the mean ±SD from a minimum of three experiments.
    Figure Legend Snippet: Telomerase regulates VEGF secretion and expression independently of telomerase activity. (A, B) VEGF secretion was assessed by ELISA following a 48-h treatment with telomerase inhibitors BIBR-1532 (10 μ M) and costunolide (10 μ M), and after 72 h of hTERT knockdown with siRNA. (C, D) Total RNAs were isolated from the cells treated in (A), and VEGF expression was analyzed by real-time PCR with GAPDH as the internal control. (E, F, G) Saos-2 cells were transiently transfected with an empty vector, hTERT-WT, and hTERT-DN. hTERT (E) and VEGF expression (F) were quantified by real-time PCR. Secreted VEGF was quantified by ELISA (G). Results were expressed as the mean ±SD from a minimum of three experiments.

    Techniques Used: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Isolation, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation

    Telomerase regulates VEGF, VEGFR1, and VEGFR2 expression. AGS (A), Caco2 (B), and HepG2/C3A (C) cells were treated with control siRNA or hTERT siRNA with or without bevacizumab, and VEGF secretion was evaluated by ELISA. (D, E) Cells were treated with BIBR-1532, costunolide, or transiently transfected with hTERT siRNA for 72h. VEGFR1 (D) and VEGFR2 (E) transcript amounts were quantified with real-time PCR. (F, G) Saos-2 cells were transiently transfected with an empty vector, hTERT-WT or hTERT-DN. VEGFR1 (F) and VEGFR2 (G) were then quantified with real-time PCR. Results were expressed as the mean ± SD from three experiments.
    Figure Legend Snippet: Telomerase regulates VEGF, VEGFR1, and VEGFR2 expression. AGS (A), Caco2 (B), and HepG2/C3A (C) cells were treated with control siRNA or hTERT siRNA with or without bevacizumab, and VEGF secretion was evaluated by ELISA. (D, E) Cells were treated with BIBR-1532, costunolide, or transiently transfected with hTERT siRNA for 72h. VEGFR1 (D) and VEGFR2 (E) transcript amounts were quantified with real-time PCR. (F, G) Saos-2 cells were transiently transfected with an empty vector, hTERT-WT or hTERT-DN. VEGFR1 (F) and VEGFR2 (G) were then quantified with real-time PCR. Results were expressed as the mean ± SD from three experiments.

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Transfection, Real-time Polymerase Chain Reaction, Plasmid Preparation

    Bevacizumab increases VEGFR1 and VEGFR2 expression. (A, B) Telomerase inhibitors were combined to bevacizumab (100 μ g/ml) and then VEGFR1 (A) and VEGFR2 (B) were quantified by real-time PCR. (C, D) Cells were transiently transfected with either control siRNA or hTERT siRNA and treated with bevacizumab (100 μ g/ml). VEGFR1 (C) and VEGFR2 (D) were quantified by real-time qPCR. (E, F) HUVECs were cultured in the presence of telomerase inhibitors for 48 h and hTERT siRNA for 72 h. The expression of VEGFR1 (E) and VEGFR2 (F) were quantified by real-time PCR. Results were expressed as the mean ± SD from three experiments.
    Figure Legend Snippet: Bevacizumab increases VEGFR1 and VEGFR2 expression. (A, B) Telomerase inhibitors were combined to bevacizumab (100 μ g/ml) and then VEGFR1 (A) and VEGFR2 (B) were quantified by real-time PCR. (C, D) Cells were transiently transfected with either control siRNA or hTERT siRNA and treated with bevacizumab (100 μ g/ml). VEGFR1 (C) and VEGFR2 (D) were quantified by real-time qPCR. (E, F) HUVECs were cultured in the presence of telomerase inhibitors for 48 h and hTERT siRNA for 72 h. The expression of VEGFR1 (E) and VEGFR2 (F) were quantified by real-time PCR. Results were expressed as the mean ± SD from three experiments.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Cell Culture

    Telomerase regulates tube formation by HUVECs. HUVECs previously treated with BIBR-1532 (10 μM), costunolide (10 μM), bevacizumab (5 ng/ml or 100 μg/ml), or hTERT siRNA were cultured on an extracellular matrix (ECM) in the presence or absence of recombinant VEGF165 (50 ng/ml). Angiogenesis began after 4 hours, and tube formation was then visualized after 6 hours, using an inverted microscope (100X).
    Figure Legend Snippet: Telomerase regulates tube formation by HUVECs. HUVECs previously treated with BIBR-1532 (10 μM), costunolide (10 μM), bevacizumab (5 ng/ml or 100 μg/ml), or hTERT siRNA were cultured on an extracellular matrix (ECM) in the presence or absence of recombinant VEGF165 (50 ng/ml). Angiogenesis began after 4 hours, and tube formation was then visualized after 6 hours, using an inverted microscope (100X).

    Techniques Used: Cell Culture, Recombinant, Inverted Microscopy

    VEGF inhibition with bevacizumab increases hTERT expression and telomerase activity. (A) Semi-quantitative RT-PCR analysis of hTERT expression from AGS, Caco2, and HepG2/C3A cells treated for 48 h with bevacizumab at 5 ng/ml and 100 μ g/ml. GAPDH was used as a loading control. (B) The amount of transcripts referred to in (A) were quantified by real-time PCR with GAPDH as an internal control. (C, D, E) Telomerase activity was detected in AGS (C), Caco2 (D), and HepG2/C3A (E). Results were expressed as the mean ± SD from three experiments.
    Figure Legend Snippet: VEGF inhibition with bevacizumab increases hTERT expression and telomerase activity. (A) Semi-quantitative RT-PCR analysis of hTERT expression from AGS, Caco2, and HepG2/C3A cells treated for 48 h with bevacizumab at 5 ng/ml and 100 μ g/ml. GAPDH was used as a loading control. (B) The amount of transcripts referred to in (A) were quantified by real-time PCR with GAPDH as an internal control. (C, D, E) Telomerase activity was detected in AGS (C), Caco2 (D), and HepG2/C3A (E). Results were expressed as the mean ± SD from three experiments.

    Techniques Used: Inhibition, Expressing, Activity Assay, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

    13) Product Images from "Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR"

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-1023-5

    Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)
    Figure Legend Snippet: Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)

    Techniques Used: Real-time Polymerase Chain Reaction, Multiplex Assay

    Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )
    Figure Legend Snippet: Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )

    Techniques Used: Sequencing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Positive Control, Multiplex Assay, Software

    14) Product Images from "No association of xenotropic murine leukemia virus-related virus with prostate cancer or chronic fatigue syndrome in Japan"

    Article Title: No association of xenotropic murine leukemia virus-related virus with prostate cancer or chronic fatigue syndrome in Japan

    Journal: Retrovirology

    doi: 10.1186/1742-4690-8-20

    Detection of XMRV RNA and DNA in viral Ab-positive samples . (A) RNA was purified from 1 mL of coculture supernatant of activated PBMCs and LNCap-FGC cells (lane 2) or 1 ml plasma (lanes 3-6). For one-step RT-PCR, 15 μl of 60 μl eluted RNA was amplified in a 25 μl volume. CFS patients C4 and C32 tested positive for XMRV Abs but C1 was negative. (B) Detection of XMRV env by TaqMan real-time PCR assay. Duplicated test samples of diluted XMRV plasmid (VP62) were amplified. The detection limit of the TaqMan real-time PCR was 4 copies/reaction determined by VP62 plasmid. (C) Duplicated test samples without template DNA in negative control (N) or with genomic DNA extracted from PBMCs of a viral Ab-positive PC patient (P24) and healthy volunteers (HV) were amplified as for (B).
    Figure Legend Snippet: Detection of XMRV RNA and DNA in viral Ab-positive samples . (A) RNA was purified from 1 mL of coculture supernatant of activated PBMCs and LNCap-FGC cells (lane 2) or 1 ml plasma (lanes 3-6). For one-step RT-PCR, 15 μl of 60 μl eluted RNA was amplified in a 25 μl volume. CFS patients C4 and C32 tested positive for XMRV Abs but C1 was negative. (B) Detection of XMRV env by TaqMan real-time PCR assay. Duplicated test samples of diluted XMRV plasmid (VP62) were amplified. The detection limit of the TaqMan real-time PCR was 4 copies/reaction determined by VP62 plasmid. (C) Duplicated test samples without template DNA in negative control (N) or with genomic DNA extracted from PBMCs of a viral Ab-positive PC patient (P24) and healthy volunteers (HV) were amplified as for (B).

    Techniques Used: Purification, Reverse Transcription Polymerase Chain Reaction, Amplification, Real-time Polymerase Chain Reaction, Plasmid Preparation, Negative Control

    15) Product Images from "Genetic variants of calcium and vitamin D metabolism in kidney stone disease"

    Article Title: Genetic variants of calcium and vitamin D metabolism in kidney stone disease

    Journal: Nature Communications

    doi: 10.1038/s41467-019-13145-x

    CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p
    Figure Legend Snippet: CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Concentration Assay, Incubation

    16) Product Images from "Effects of Trichostatin A on the Chondrogenesis from Human Mesenchymal Stem Cells"

    Article Title: Effects of Trichostatin A on the Chondrogenesis from Human Mesenchymal Stem Cells

    Journal: Tissue Engineering and Regenerative Medicine

    doi: 10.1007/s13770-017-0041-6

    TSA inhibits chondrogenic differentiation of hBMMSCs. A Representative images of chondrogenic pellets after day 21 ( scale = 1 mm), DNA quantity, and GAG per DNA. B qRT-PCR of chondrogenic markers in the chondrogenic pellets. C Hematoxylin and eosin (H E), Alcian blue, and Safranin-O staining of chondrogenic pellets ( scale bars = 0.3 mm). n = 3, * p
    Figure Legend Snippet: TSA inhibits chondrogenic differentiation of hBMMSCs. A Representative images of chondrogenic pellets after day 21 ( scale = 1 mm), DNA quantity, and GAG per DNA. B qRT-PCR of chondrogenic markers in the chondrogenic pellets. C Hematoxylin and eosin (H E), Alcian blue, and Safranin-O staining of chondrogenic pellets ( scale bars = 0.3 mm). n = 3, * p

    Techniques Used: Quantitative RT-PCR, Staining

    qRT-PCR validation of TSA-induced gene expression changes. A Col10Al, TGF-β3, and SOX9 were downregulated, and B BMP4 and FGFR3 were upregulated in TSA-treated pellets; n = 3, * p
    Figure Legend Snippet: qRT-PCR validation of TSA-induced gene expression changes. A Col10Al, TGF-β3, and SOX9 were downregulated, and B BMP4 and FGFR3 were upregulated in TSA-treated pellets; n = 3, * p

    Techniques Used: Quantitative RT-PCR, Expressing

    17) Product Images from "A genetically engineered microRNA-34a prodrug demonstrates anti-tumor activity in a canine model of osteosarcoma"

    Article Title: A genetically engineered microRNA-34a prodrug demonstrates anti-tumor activity in a canine model of osteosarcoma

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0209941

    Bioengineered tRNA/miR-34a prodrug was processed to mature miR-34a in canine OS cell lines. Relative expression levels of mature miR-34a in cells 24 hours after treatment (10 nM tRNA/MSA or tRNA/miR-34a) were analyzed by qRT-PCR and normalized versus geometric mean of housekeeping genes Rps5 , Gapdh and Hnrnph1 . Data (mean±SEM) were analyzed independently analyzed by Student’s t -test.
    Figure Legend Snippet: Bioengineered tRNA/miR-34a prodrug was processed to mature miR-34a in canine OS cell lines. Relative expression levels of mature miR-34a in cells 24 hours after treatment (10 nM tRNA/MSA or tRNA/miR-34a) were analyzed by qRT-PCR and normalized versus geometric mean of housekeeping genes Rps5 , Gapdh and Hnrnph1 . Data (mean±SEM) were analyzed independently analyzed by Student’s t -test.

    Techniques Used: Expressing, Quantitative RT-PCR

    18) Product Images from "Genetic variants of calcium and vitamin D metabolism in kidney stone disease"

    Article Title: Genetic variants of calcium and vitamin D metabolism in kidney stone disease

    Journal: Nature Communications

    doi: 10.1038/s41467-019-13145-x

    CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p
    Figure Legend Snippet: CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Concentration Assay, Incubation

    19) Product Images from "Genetic variants of calcium and vitamin D metabolism in kidney stone disease"

    Article Title: Genetic variants of calcium and vitamin D metabolism in kidney stone disease

    Journal: Nature Communications

    doi: 10.1038/s41467-019-13145-x

    CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p
    Figure Legend Snippet: CaSR-mediated SRE responses following DGKD knockdown and effect of cinacalcet treatment in HEK-CaSR-SRE cells. a Relative expression of DGKD , as assessed by quantitative real-time PCR of HEK-CaSR-SRE cells treated with scrambled (WT) or DGKD (DGKD-KD) siRNA and used for SRE experiments. Samples were normalized to a geometric mean of four housekeeper genes: PGK1 , GAPDH , TUB1A , CDNK1B . n = 8 biologically independent transfections. b Representative western blot of lysates from HEK-CaSR cells treated with scrambled or DGKD siRNA and used for SRE experiments. α−Tubulin was used as a loading control. c Relative expression of DGKD, as assessed by densitometry of western blots from cells treated with scrambled or DGKD siRNA demonstrating a ~50% reduction in expression of DGKD following treatment with DGKD siRNA. Samples were normalized to PGK1. n = 6 biologically independent transfections for WT, n = 9 biologically independent transfections for DGKD-KD. d SRE responses of HEK-CaSR-SRE cells in response to changes in extracellular calcium concentration. Cells were treated with scrambled (WT) or DGKD (DGKD-KD) siRNA. The responses ± SEM are shown for n = 8 biologically independent transfections for WT and DGKD-KD cells and n = 4 biologically independent transfections for DGKD-KD + 5 nM cinacalcet cells. Treatment with DGKD siRNA led to a reduction in maximal response (red line) compared to cells treated with scrambled siRNA (black line). This loss-of-function could be rectified by treatment with 5 nM cinacalcet (blue line). Post desensitization points are shown but were not included in the analysis (gray, light red, and light blue). e Mean maximal responses with SEM of cells treated with scrambled siRNA (WT, black), DGKD siRNA (DGKD-KD, red) and DGKD siRNA incubated with 5 nM cinacalcet (blue). Statistical comparisons of maximal response were undertaken using F test. Student’s t-tests were used to compare relative expression. Two-way ANOVA was used to compare points on dose response curve with reference to WT. Data are shown as mean ± SEM with ** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Concentration Assay, Incubation

    20) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    21) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    22) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.
    Figure Legend Snippet: Comparison of cytomegalovirus (CMV, N=38) and Epstein-Barr virus (EBV, N=17) results using the QIAsymphony RGQ and QIA-cube systems. (A) Passing-Bablok regression for CMV detection. (B) Bland-Altman plot for CMV detection. (C) Passing-Bablok regression for EBV detection. (D) Bland-Altman plot for EBV detection. In the regression plot, the solid line indicates the regression line and dashed lines indicate 95% confidence interval. In the Bland-Altman plot, the bold line indicates the mean difference between values, the dashed lines indicate 95% confidence interval, and the solid lines indicate mean difference ±1.96 standard deviation.

    Techniques Used: Standard Deviation

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    23) Product Images from "A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance"

    Article Title: A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance

    Journal: Scientific Reports

    doi: 10.1038/srep42892

    Melting curve analysis of the targeted eight poxviruses using different PCR platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).
    Figure Legend Snippet: Melting curve analysis of the targeted eight poxviruses using different PCR platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).

    Techniques Used: Polymerase Chain Reaction, Amplification, Infection

    24) Product Images from "Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR"

    Article Title: Identification and quantification of virulence factors of enterotoxigenic Escherichia coli by high-resolution melting curve quantitative PCR

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-1023-5

    Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)
    Figure Legend Snippet: Scatter plot and regression analysis between the Log 10 gene copy number of F18 fimbriae gene measured by qPCR and multiplex HRM-qPCR methods ( n = 14)

    Techniques Used: Real-time Polymerase Chain Reaction, Multiplex Assay

    Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )
    Figure Legend Snippet: Melting curve of target sequence from positive controls by individual HRM-qPCR ( top ), melting curve of PCR products amplified from a mixed positive control including five different fimbriae gene sequence by multiplex HRM-qPCR ( bottom, dotted ) and the corresponding reprocessed melting curve by PeakFit software ( bottom, solid )

    Techniques Used: Sequencing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Positive Control, Multiplex Assay, Software

    25) Product Images from "The Transcription Factor MEF2C Negatively Controls Angiogenic Sprouting of Endothelial Cells Depending on Oxygen"

    Article Title: The Transcription Factor MEF2C Negatively Controls Angiogenic Sprouting of Endothelial Cells Depending on Oxygen

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0101521

    A2M mediates the inhibitive effect of MEF2C on sprouting. (a) Inhibition of sprouting by purified A2M. Increasing concentrations of commercially available A2M were added to the spheroid sprouting assay and the effects on total sprout length scored. Data are displayed as mean values ± SEM as calculated from three experiments. (b) A2M mRNA expression is strongly reduced upon shRNA mediated knockdown. HUVEC were transduced with LV.shA2M or LV.shcon for 48 hours. Total RNA was isolated, subjected to cDNA synthesis and analyzed by realtime RT-PCR. Values were normalized to beta-2-microglobulin mRNA as internal standard and displayed as mean ±SD. One representative experiment of 3 performed in triplicates is shown (c) Knockdown of A2M restores sprouting activity in Ad.MEF2C transduced HUVEC. Cells were first transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 6 hours before they were used to form spheroids for the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was scored. Data displayed are mean values ± SEM calculated from three experiments. (d) Knockdown of A2M reduces the inhibitory activity in conditioned media from Ad.MEF2C transduced cells. HUVEC were transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 8 hours. Then medium was changed to serum-free Opti-MEM medium, supernatants were harvested after 48 hours, concentrated by diafiltration and added to the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was assessed. Values depicted are mean values ± SEM calculated from three independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. n.s. not significant, ***p
    Figure Legend Snippet: A2M mediates the inhibitive effect of MEF2C on sprouting. (a) Inhibition of sprouting by purified A2M. Increasing concentrations of commercially available A2M were added to the spheroid sprouting assay and the effects on total sprout length scored. Data are displayed as mean values ± SEM as calculated from three experiments. (b) A2M mRNA expression is strongly reduced upon shRNA mediated knockdown. HUVEC were transduced with LV.shA2M or LV.shcon for 48 hours. Total RNA was isolated, subjected to cDNA synthesis and analyzed by realtime RT-PCR. Values were normalized to beta-2-microglobulin mRNA as internal standard and displayed as mean ±SD. One representative experiment of 3 performed in triplicates is shown (c) Knockdown of A2M restores sprouting activity in Ad.MEF2C transduced HUVEC. Cells were first transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 6 hours before they were used to form spheroids for the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was scored. Data displayed are mean values ± SEM calculated from three experiments. (d) Knockdown of A2M reduces the inhibitory activity in conditioned media from Ad.MEF2C transduced cells. HUVEC were transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 8 hours. Then medium was changed to serum-free Opti-MEM medium, supernatants were harvested after 48 hours, concentrated by diafiltration and added to the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was assessed. Values depicted are mean values ± SEM calculated from three independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. n.s. not significant, ***p

    Techniques Used: Inhibition, Purification, Expressing, shRNA, Transduction, Isolation, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Infection, Diafiltration Assay

    Alpha-2-macroglobulin (A2M) is upregulated by MEF2C in HUVEC on mRNA and protein level. (a) A2M and MEF2C mRNA levels. HUVEC were transduced for 8, 16, 24 and 32 hours with Ad.MEF2C or Ad.con using MOI of 10 or left without infection (0 hours value). RNAs were isolated from the cells and A2M as well as MEF2C mRNA levels were determined by realtime RT-PCR as described in detail in the Methods section. Fold induction levels are shown as mean values ± SD calculated from triplicates of one representative experiment out of three independent experiments performed. Obtained values were normalized to beta-2-microglobulin mRNA as internal standard. (b) A2M protein. HUVEC were infected with Ad.MEF2C, Ad.con or left without infection for 8 hours, then medium was changed to serum-free Opti-MEM medium and supernatants were harvested after 48 hours. A2M secreted into supernatants was determined using a commercial A2M ELISA kit and a standard curve obtained with purified A2M. The mean concentration of A2M ± SD as calculated from three HUVEC batches analyzed is shown. (c) Supernatants of MEF2C virus transduced HUVEC inhibit sprouting. HUVEC were transduced with Ad.con or Ad.MEF2C or left uninduced for 24 hours, then medium was changed to Opti-mem, after 48 hours supernatants were collected and further concentrated by diafiltration as described in the Methods section. These conditioned media were added to the sprouting assay (50–70 µl to a 500 µl assay) and VEGF-A/bFGF-induced as well as basal sprout formation was scored . Data are displayed as mean values ± SEM as calculated from 3 independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. ***p
    Figure Legend Snippet: Alpha-2-macroglobulin (A2M) is upregulated by MEF2C in HUVEC on mRNA and protein level. (a) A2M and MEF2C mRNA levels. HUVEC were transduced for 8, 16, 24 and 32 hours with Ad.MEF2C or Ad.con using MOI of 10 or left without infection (0 hours value). RNAs were isolated from the cells and A2M as well as MEF2C mRNA levels were determined by realtime RT-PCR as described in detail in the Methods section. Fold induction levels are shown as mean values ± SD calculated from triplicates of one representative experiment out of three independent experiments performed. Obtained values were normalized to beta-2-microglobulin mRNA as internal standard. (b) A2M protein. HUVEC were infected with Ad.MEF2C, Ad.con or left without infection for 8 hours, then medium was changed to serum-free Opti-MEM medium and supernatants were harvested after 48 hours. A2M secreted into supernatants was determined using a commercial A2M ELISA kit and a standard curve obtained with purified A2M. The mean concentration of A2M ± SD as calculated from three HUVEC batches analyzed is shown. (c) Supernatants of MEF2C virus transduced HUVEC inhibit sprouting. HUVEC were transduced with Ad.con or Ad.MEF2C or left uninduced for 24 hours, then medium was changed to Opti-mem, after 48 hours supernatants were collected and further concentrated by diafiltration as described in the Methods section. These conditioned media were added to the sprouting assay (50–70 µl to a 500 µl assay) and VEGF-A/bFGF-induced as well as basal sprout formation was scored . Data are displayed as mean values ± SEM as calculated from 3 independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. ***p

    Techniques Used: Infection, Isolation, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Transduction, Diafiltration Assay

    26) Product Images from "Transcriptional profiling analysis and functional prediction of long noncoding RNAs in cancer"

    Article Title: Transcriptional profiling analysis and functional prediction of long noncoding RNAs in cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.6993

    qRT-PCR validation of lncRNA and mRNA expression in samples from external patients ( A ) Expression of common DE-mRNAs/lncRNAs (upper panel representing a mRNA and lower panel representing a lncRNA). ( B ) Boxplots represent common DE-mRNAs/lncRNAs' expression based on the microarray data. (upper panel representing a mRNA and lower panel representing a lncRNA).
    Figure Legend Snippet: qRT-PCR validation of lncRNA and mRNA expression in samples from external patients ( A ) Expression of common DE-mRNAs/lncRNAs (upper panel representing a mRNA and lower panel representing a lncRNA). ( B ) Boxplots represent common DE-mRNAs/lncRNAs' expression based on the microarray data. (upper panel representing a mRNA and lower panel representing a lncRNA).

    Techniques Used: Quantitative RT-PCR, Expressing, Microarray

    27) Product Images from "Microbial Typing by Machine Learned DNA Melt Signatures"

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures

    Journal: Scientific Reports

    doi: 10.1038/srep42097

    Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.
    Figure Legend Snippet: Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.

    Techniques Used: Sequencing, Polymerase Chain Reaction, Marker, Agarose Gel Electrophoresis, Molecular Weight

    28) Product Images from "Microbial Typing by Machine Learned DNA Melt Signatures"

    Article Title: Microbial Typing by Machine Learned DNA Melt Signatures

    Journal: Scientific Reports

    doi: 10.1038/srep42097

    Workflow of our ITS HRM assay. The ITS HRM assay is performed on DNA extracted from clinical samples such as positive blood culture samples. Our novel classification algorithm allows generated melt curves to identify the bacterial pathogens by matching against a reference database. Same-colored dots represent replicates from the same species classified within their boundaries and black dots outside all boundaries represent replicates from species that is not in our reference database, whereas the unknown species in the test sample (X) is identified by the classification algorithm.
    Figure Legend Snippet: Workflow of our ITS HRM assay. The ITS HRM assay is performed on DNA extracted from clinical samples such as positive blood culture samples. Our novel classification algorithm allows generated melt curves to identify the bacterial pathogens by matching against a reference database. Same-colored dots represent replicates from the same species classified within their boundaries and black dots outside all boundaries represent replicates from species that is not in our reference database, whereas the unknown species in the test sample (X) is identified by the classification algorithm.

    Techniques Used: HRM Assay, Generated

    Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.
    Figure Legend Snippet: Heteroduplex analysis of E. coli ITS. ( a ) Clustal Omega Multiple Sequence Alignment of E. coli ATCC 25922 (GenBank Accession number CP009072) ITS short (361 bp) and ITS long (453 bp) sequences. ( b ) ITS homoduplex-heteroduplex profiles obtained after 20 (1), 25 (2), 30 (3), 35 (4) and 40 (5) number of PCR cycles. Slow migrating bands (at 800 and 1000 bp) were visible starting from cycle number 25, suggesting the heteroduplex nature of the bands. Expected homoduplex bands were at 540 bp (361 bp ITS short + 179 bp of 3′ of 16S and 38 bp of 5′ 23S), and 632 bp (453 bp + 179 bp). Lane M contains 100-bp DNA marker. ( c ) Agarose gel electrophoresis showing E. coli ITS PCR products treated (2) and untreated (1) with mung bean nuclease, an enzyme that recognizes and cleaves single stranded DNA, even when it is located in double-stranded DNA products. The loss of the higher molecular weight bands confirms the heteroduplex nature of the bands, leaving the true homoduplexes. Lane M contains 100-bp DNA marker. ( d ) ITS HRM analysis on 20 colonies resulted in 2 distinct melt curve groups (ITS short and ITS long). Combinations of the amplicons from each group did not recreate the original E. coli melt curve.

    Techniques Used: Sequencing, Polymerase Chain Reaction, Marker, Agarose Gel Electrophoresis, Molecular Weight

    29) Product Images from "OsWRKY67 Plays a Positive Role in Basal and XA21-Mediated Resistance in Rice"

    Article Title: OsWRKY67 Plays a Positive Role in Basal and XA21-Mediated Resistance in Rice

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2017.02220

    Disease resistance phenotype of OsWRKY67 RNAi lines to Xoo isolates. (A) Schematic representation of the OsWRKY67 RNAi construct. Sense and antisense boxes indicate a unique 359 bp fragment of OsWRKY67 and antisense sequence (Top). Suppression of OsWRKY67 transcripts were validated by qPCR. OsUbq5 is a PCR control. Kit-XA21 is a transformation background genotype; NT is a segregant nontransgene-bearing wild type. (B) Representative leaves 12 days after inoculation with Xoo PXO99. Yellow asterisks indicate bottoms of lesions. (C) Disease lesion lengths measured 12 days after inoculation with Xoo PXO99. (D) Representative leaves 12 days after inoculation with Xoo KXO85. (E) Disease lesion lengths measured 12 days after inoculation with Xoo KXO85. Data are represented as means ± SD. Asterisks represent statistical significance with Student's t -test, p
    Figure Legend Snippet: Disease resistance phenotype of OsWRKY67 RNAi lines to Xoo isolates. (A) Schematic representation of the OsWRKY67 RNAi construct. Sense and antisense boxes indicate a unique 359 bp fragment of OsWRKY67 and antisense sequence (Top). Suppression of OsWRKY67 transcripts were validated by qPCR. OsUbq5 is a PCR control. Kit-XA21 is a transformation background genotype; NT is a segregant nontransgene-bearing wild type. (B) Representative leaves 12 days after inoculation with Xoo PXO99. Yellow asterisks indicate bottoms of lesions. (C) Disease lesion lengths measured 12 days after inoculation with Xoo PXO99. (D) Representative leaves 12 days after inoculation with Xoo KXO85. (E) Disease lesion lengths measured 12 days after inoculation with Xoo KXO85. Data are represented as means ± SD. Asterisks represent statistical significance with Student's t -test, p

    Techniques Used: Construct, Sequencing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Transformation Assay

    Disease resistance phenotype of OsWRKY67 activation mutants. (A) Schematic representation of OsWRKY67 activation mutant alleles OsW67-D1 and OsW67-D2 with T-DNA insertion carrying a 4 × 35S enhancer on the diagram of the OsWRKY67 gene. Gray boxes and black lines represent exons and introns, respectively (Top). Levels of OsWRKY67 expression were validated by qPCR (Bottom). WT is a wild type control cultivar, Dongjin. OsUbq5 is a PCR control. (B) Representative leaves 9 days after inoculation with M. oryzae PO6-6. (C) Disease lesion lengths measured 9 days after inoculation with M. oryzae PO6-6. (D) Representative leaves 12 days after inoculation with Xoo PXO99. Yellow asterisks indicate bottoms of lesions. (E) Disease lesion lengths measured 12 days after inoculation with Xoo PXO99. Data are represented as means ± SD. Asterisks represent statistical significance with Student's t -test, p
    Figure Legend Snippet: Disease resistance phenotype of OsWRKY67 activation mutants. (A) Schematic representation of OsWRKY67 activation mutant alleles OsW67-D1 and OsW67-D2 with T-DNA insertion carrying a 4 × 35S enhancer on the diagram of the OsWRKY67 gene. Gray boxes and black lines represent exons and introns, respectively (Top). Levels of OsWRKY67 expression were validated by qPCR (Bottom). WT is a wild type control cultivar, Dongjin. OsUbq5 is a PCR control. (B) Representative leaves 9 days after inoculation with M. oryzae PO6-6. (C) Disease lesion lengths measured 9 days after inoculation with M. oryzae PO6-6. (D) Representative leaves 12 days after inoculation with Xoo PXO99. Yellow asterisks indicate bottoms of lesions. (E) Disease lesion lengths measured 12 days after inoculation with Xoo PXO99. Data are represented as means ± SD. Asterisks represent statistical significance with Student's t -test, p

    Techniques Used: Activation Assay, Mutagenesis, Expressing, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    30) Product Images from "Multiple Targets of Nitric Oxide in the Tricarboxylic Acid (TCA) Cycle of Salmonella enterica Serovar Typhimurium"

    Article Title: Multiple Targets of Nitric Oxide in the Tricarboxylic Acid (TCA) Cycle of Salmonella enterica Serovar Typhimurium

    Journal: Cell host & microbe

    doi: 10.1016/j.chom.2011.06.004

    NO· creates a regulatory block that inhibits the reductive branch of the TCA cycle A. Expression levels of sdhC or frdA as determined by Q-RT PCR of three independent RNA samples from WT and isogenic Δ fnr S. Typhimurium strain 14028s either untreated (blue columns) or exposed to NO· (red columns). Transcript levels relative to rpoD were determined by a modified ΔΔC t method as previously described. B. Expressing the reductive TCA cycle in the presence of NO· restores MK prototrophy to S. Typhimurium. The frdABCD genes from 14028s were cloned and expressed from an IPTG-inducible promoter (p trc ). IPTG was added at 100 μM for each growth curve. M9 Glc medium was either unsupplemented (None), supplemented with all 20 amino acids (All), or with a mixture of amino acids lacking both Met and Lys (−MK). pTrc99a was used as a vector control. A Δ sdh mutation was introduced to prevent futile cycling by high expression of both SDH and FRD activities.
    Figure Legend Snippet: NO· creates a regulatory block that inhibits the reductive branch of the TCA cycle A. Expression levels of sdhC or frdA as determined by Q-RT PCR of three independent RNA samples from WT and isogenic Δ fnr S. Typhimurium strain 14028s either untreated (blue columns) or exposed to NO· (red columns). Transcript levels relative to rpoD were determined by a modified ΔΔC t method as previously described. B. Expressing the reductive TCA cycle in the presence of NO· restores MK prototrophy to S. Typhimurium. The frdABCD genes from 14028s were cloned and expressed from an IPTG-inducible promoter (p trc ). IPTG was added at 100 μM for each growth curve. M9 Glc medium was either unsupplemented (None), supplemented with all 20 amino acids (All), or with a mixture of amino acids lacking both Met and Lys (−MK). pTrc99a was used as a vector control. A Δ sdh mutation was introduced to prevent futile cycling by high expression of both SDH and FRD activities.

    Techniques Used: Blocking Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Modification, Clone Assay, Gas Chromatography, Plasmid Preparation, Mutagenesis

    31) Product Images from "The Expression of Activating Receptor Gene of Natural Killer Cells (KLRC3) in Patients with 
Type 1 Diabetes Mellitus (T1DM)"

    Article Title: The Expression of Activating Receptor Gene of Natural Killer Cells (KLRC3) in Patients with 
Type 1 Diabetes Mellitus (T1DM)

    Journal: Oman Medical Journal

    doi: 10.5001/omj.2017.60

    KLRC3 gene expression in the three study groups.
    Figure Legend Snippet: KLRC3 gene expression in the three study groups.

    Techniques Used: Expressing

    Melting curve for KLRC3 gene.
    Figure Legend Snippet: Melting curve for KLRC3 gene.

    Techniques Used:

    Amplification plot curves for KLRC3 gene expression.
    Figure Legend Snippet: Amplification plot curves for KLRC3 gene expression.

    Techniques Used: Amplification, Expressing

    32) Product Images from "MicroRNA-145 Antagonism Reverses TGF-β Inhibition of F508del CFTR Correction in Airway Epithelia"

    Article Title: MicroRNA-145 Antagonism Reverses TGF-β Inhibition of F508del CFTR Correction in Airway Epithelia

    Journal: American Journal of Respiratory and Critical Care Medicine

    doi: 10.1164/rccm.201704-0732OC

    TGF (transforming growth factor)-β stimulates microRNA (miR)-145 in cystic fibrosis (CF) BAL fluid and airway epithelial cells. ( A ) Increased miR-145 in human CF BAL–derived exosomes ( n = 7) compared with non-CF ( n = 5). ( B and C ) TGF-β transcription and signaling (plasminogen activatory 1 [PAI-1] expression) are increased in human CF BAL–derived exosomes ( n = 6) compared with non-CF ( n = 4) samples. ( D ) Baseline expression of TGF-β transcription and signaling (PAI-1) is increased in CF airway epithelial cells ( n = 3) without stimulation. ( E ) Addition of exogenous TGF-β (5 ng/ml) further doubles miR-145 in non-CF and CF epithelial cells compared with DMSO control ( n = 3). All quantitative PCR experiments were repeated at least three times. Data points represent summary data from each donor. Normalized data expression is presented as median value with interquartile range. DMSO = dimethyl sulfoxide. * P
    Figure Legend Snippet: TGF (transforming growth factor)-β stimulates microRNA (miR)-145 in cystic fibrosis (CF) BAL fluid and airway epithelial cells. ( A ) Increased miR-145 in human CF BAL–derived exosomes ( n = 7) compared with non-CF ( n = 5). ( B and C ) TGF-β transcription and signaling (plasminogen activatory 1 [PAI-1] expression) are increased in human CF BAL–derived exosomes ( n = 6) compared with non-CF ( n = 4) samples. ( D ) Baseline expression of TGF-β transcription and signaling (PAI-1) is increased in CF airway epithelial cells ( n = 3) without stimulation. ( E ) Addition of exogenous TGF-β (5 ng/ml) further doubles miR-145 in non-CF and CF epithelial cells compared with DMSO control ( n = 3). All quantitative PCR experiments were repeated at least three times. Data points represent summary data from each donor. Normalized data expression is presented as median value with interquartile range. DMSO = dimethyl sulfoxide. * P

    Techniques Used: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction

    MicroRNA (miRNA) expression profiles in human airway epithelial cells. ( A ) miRNA-145 (miR-145) predicted binding site on 3′-untranslated region (UTR) of CFTR (cystic fibrosis transmembrane conductance regulator) mRNA that is highly conserved across species (TargetScan 7.1). ( B ) Luciferase reporter activity for miR-145 binding to CFTR 3′-UTR was measured in CFBE41o – cell lines. Each data point represents summary data from three independent experiments ( n = 3). ( C ) Expression profiles of CFTR-specific miRNAs (miR-145, miR-101, and miR-494) in non–cystic fibrosis (CF) and CF primary airway epithelial cells. Each miRNA data point represents summary data from three biological replicates ( n = 3 donors). ( D ) Increased miR-145 in F508del CFBE cell line ( n = 3). All experiments were repeated at least three times. Quantitative PCR data were analyzed by the ΔΔ C t method. Data are presented as the median of normalized expression (to U6 small nuclear RNA) with interquartile range. WT = wild type. * P
    Figure Legend Snippet: MicroRNA (miRNA) expression profiles in human airway epithelial cells. ( A ) miRNA-145 (miR-145) predicted binding site on 3′-untranslated region (UTR) of CFTR (cystic fibrosis transmembrane conductance regulator) mRNA that is highly conserved across species (TargetScan 7.1). ( B ) Luciferase reporter activity for miR-145 binding to CFTR 3′-UTR was measured in CFBE41o – cell lines. Each data point represents summary data from three independent experiments ( n = 3). ( C ) Expression profiles of CFTR-specific miRNAs (miR-145, miR-101, and miR-494) in non–cystic fibrosis (CF) and CF primary airway epithelial cells. Each miRNA data point represents summary data from three biological replicates ( n = 3 donors). ( D ) Increased miR-145 in F508del CFBE cell line ( n = 3). All experiments were repeated at least three times. Quantitative PCR data were analyzed by the ΔΔ C t method. Data are presented as the median of normalized expression (to U6 small nuclear RNA) with interquartile range. WT = wild type. * P

    Techniques Used: Expressing, Binding Assay, Luciferase, Activity Assay, Real-time Polymerase Chain Reaction

    33) Product Images from "SWI/SNF interacts with cleavage and polyadenylation factors and facilitates pre-mRNA 3′ end processing"

    Article Title: SWI/SNF interacts with cleavage and polyadenylation factors and facilitates pre-mRNA 3′ end processing

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gky438

    The cleavage of CG5174 and Hat1 pre-mRNAs is regulated by dBRM. ( A ) Drawing showing the position of the primers used for RT-qPCR in the cleavage assay. The thick box represents the 3′ UTR. The thin stretch represents genomic sequences downstream of the cleavage site (CS). The relative amount of uncleaved pre-mRNA was quantified using the uncleaved primers and normalized to the amount of 3′ UTR (using the 3′ UTR primers). ( B ) S2 cells were treated with dsRNA complementary to either dBRM or GFP (control) for 48 h. The relative levels of uncleaved pre-mRNAs were quantified by RT-qPCR, and normalized to the 3′ UTR levels of each transcript. The bars show the average ratio between dsBRM and dsGFP samples. The error bars are standard deviations from three biological replicates. ( C ) Stably transfected S2 cells were treated with 200 μM CuSO 4 for 24 h to induce the expression of recombinant dBRM. The relative levels of uncleaved pre-mRNAs were quantified by RT-qPCR as in C. The error bars are standard deviations from three biological replicates. ( D ) dBRM occupancy in four selected genes visualized in the IGV browser. The Y-axis shows the enrichment of the ChIP signal relative to the input. The bracketed numbers on each panel show the y-axis values. CG13919 is shown as an example of a gene that is not occupied by dBRM. The dBRM ChIP-seq data was from Jordán-Pla et al. ). In B and C, one sample t -tests were used for statistical testing. Significant probability ( p ) values are shown in the figure.
    Figure Legend Snippet: The cleavage of CG5174 and Hat1 pre-mRNAs is regulated by dBRM. ( A ) Drawing showing the position of the primers used for RT-qPCR in the cleavage assay. The thick box represents the 3′ UTR. The thin stretch represents genomic sequences downstream of the cleavage site (CS). The relative amount of uncleaved pre-mRNA was quantified using the uncleaved primers and normalized to the amount of 3′ UTR (using the 3′ UTR primers). ( B ) S2 cells were treated with dsRNA complementary to either dBRM or GFP (control) for 48 h. The relative levels of uncleaved pre-mRNAs were quantified by RT-qPCR, and normalized to the 3′ UTR levels of each transcript. The bars show the average ratio between dsBRM and dsGFP samples. The error bars are standard deviations from three biological replicates. ( C ) Stably transfected S2 cells were treated with 200 μM CuSO 4 for 24 h to induce the expression of recombinant dBRM. The relative levels of uncleaved pre-mRNAs were quantified by RT-qPCR as in C. The error bars are standard deviations from three biological replicates. ( D ) dBRM occupancy in four selected genes visualized in the IGV browser. The Y-axis shows the enrichment of the ChIP signal relative to the input. The bracketed numbers on each panel show the y-axis values. CG13919 is shown as an example of a gene that is not occupied by dBRM. The dBRM ChIP-seq data was from Jordán-Pla et al. ). In B and C, one sample t -tests were used for statistical testing. Significant probability ( p ) values are shown in the figure.

    Techniques Used: Quantitative RT-PCR, Cleavage Assay, Genomic Sequencing, Stable Transfection, Transfection, Expressing, Recombinant, Chromatin Immunoprecipitation, Proximity Ligation Assay

    34) Product Images from "Inflammation promotes adipocyte lipolysis via IRE1 kinase"

    Article Title: Inflammation promotes adipocyte lipolysis via IRE1 kinase

    Journal: bioRxiv

    doi: 10.1101/2020.04.07.030148

    The tyrosine kinase inhibitors ponatinib and imatinib do not alter the IRE1-mediated lipolysis in 3T3-L1 adipocytes. A, B, D) Glycerol release rates were calculated in adipocytes treated with vehicle (control), 10 µg/mL PGN, 1 µM thapsigargin, 2 µM isoproterenol, 500 ng/mL LPS, 10 ng/mL TNF, or 20 ng/mL Il6, as indicated (N=8-10). Cells were treated with ponatinib (0.1 µM) or imatinib (5 µM) for 1 hour prior to ligand addition. C) Relative transcript levels of spliced XBP1 (sXBP1) in 3T3-L1 adipocytes treated with 0.25µM Thapsigargin for 3 h with or without 5 µM imatinib. E) NF-κβ activation in HEK-Blue™ NOD1 cells treated with vehicle, 10 µg/mL PGN or 10 ng/mL TNF in the presence or absence of 0.1 µM ponatinib or 5 µM imatinib (N=8-9). F-G) Il6 secretion in 3T3-L1 adipocytes after 48 hours of incubation with or without 0.1 µM ponatinib (F) or 5 µM imatinib (G) and 10 µg/mL PGN, 1 µM thapsigargin, 2 µM isoproterenol, 500 ng/mL LPS, or 10 ng/mL TNF, as indicated (N=7-10). Values are mean ± SEM. Statistical significance was measured as p
    Figure Legend Snippet: The tyrosine kinase inhibitors ponatinib and imatinib do not alter the IRE1-mediated lipolysis in 3T3-L1 adipocytes. A, B, D) Glycerol release rates were calculated in adipocytes treated with vehicle (control), 10 µg/mL PGN, 1 µM thapsigargin, 2 µM isoproterenol, 500 ng/mL LPS, 10 ng/mL TNF, or 20 ng/mL Il6, as indicated (N=8-10). Cells were treated with ponatinib (0.1 µM) or imatinib (5 µM) for 1 hour prior to ligand addition. C) Relative transcript levels of spliced XBP1 (sXBP1) in 3T3-L1 adipocytes treated with 0.25µM Thapsigargin for 3 h with or without 5 µM imatinib. E) NF-κβ activation in HEK-Blue™ NOD1 cells treated with vehicle, 10 µg/mL PGN or 10 ng/mL TNF in the presence or absence of 0.1 µM ponatinib or 5 µM imatinib (N=8-9). F-G) Il6 secretion in 3T3-L1 adipocytes after 48 hours of incubation with or without 0.1 µM ponatinib (F) or 5 µM imatinib (G) and 10 µg/mL PGN, 1 µM thapsigargin, 2 µM isoproterenol, 500 ng/mL LPS, or 10 ng/mL TNF, as indicated (N=7-10). Values are mean ± SEM. Statistical significance was measured as p

    Techniques Used: Activation Assay, Incubation

    Inflammatory ligands stimulate lipolysis via IRE1 kinase not RNase activity. A-C) Glycerol release rates (µM/h) in 3T3-L1 adipocytes treated with increasing concentrations of the IRE1 inhibitor KirA6 (A; N=4), the IRE1 RNase inhibitor KirA8 (B; N=16), or the IRE1 RNase inhibitor 4µ8C (C; N=12-16). D) Expression of spliced Xbp1 in 3T3-L1 adipocytes treated with 1 μM Thapsigargin for 3 hours with and without 10 μM KirA8. E) Glycerol release rates in differentiated 3T3-L1 adipocytes treated with or without 1 µM KirA6 (IRE1 inhibitor) and one of vehicle (mock-treatment), 10 µg/mL FK565 (PGN), 1 µM thapsigargin (ER stress activator), 2 µM isoproterenol (β-adrenergic agonist), 500 ng/mL LPS (TLR4 activator), or 10 ng/mL TNF, as indicated (N=12-14). F) Glycerol release rates in 3T3-L1 adipocytes treated with 1 μM KirA6 and 0.5 mM 8-Br-cAMP (N=10-12). G) Phosphorylation of IRE1 was measured in 3T3-L1 adipocytes after stimulation with vehicle, PGN (10 µg/mL), or thapsigargin (1 µM) for 3 hours, with or without 1 µM KirA6 (N=4-6). Blots were stripped and re-probed for total IRE1. H) Quantification of Western blots for the ratio between phosphorylated IRE1 and total IRE1, expressed relative to the basal control sample. I-K) RNA isolated from 3T3-L1 adipocytes was treated for 3 hours with vehicle, FK565 (PGN, 10 µg/mL), thapsigargin (1 µM), or isoproterenol (2 µM) in the absence or presence of KirA6 (1 µM). Expression of BiP (I), CHOP (J), and spliced Xbp1(K) (N=12). Values are mean ± SEM. Statistical significance was measured as p
    Figure Legend Snippet: Inflammatory ligands stimulate lipolysis via IRE1 kinase not RNase activity. A-C) Glycerol release rates (µM/h) in 3T3-L1 adipocytes treated with increasing concentrations of the IRE1 inhibitor KirA6 (A; N=4), the IRE1 RNase inhibitor KirA8 (B; N=16), or the IRE1 RNase inhibitor 4µ8C (C; N=12-16). D) Expression of spliced Xbp1 in 3T3-L1 adipocytes treated with 1 μM Thapsigargin for 3 hours with and without 10 μM KirA8. E) Glycerol release rates in differentiated 3T3-L1 adipocytes treated with or without 1 µM KirA6 (IRE1 inhibitor) and one of vehicle (mock-treatment), 10 µg/mL FK565 (PGN), 1 µM thapsigargin (ER stress activator), 2 µM isoproterenol (β-adrenergic agonist), 500 ng/mL LPS (TLR4 activator), or 10 ng/mL TNF, as indicated (N=12-14). F) Glycerol release rates in 3T3-L1 adipocytes treated with 1 μM KirA6 and 0.5 mM 8-Br-cAMP (N=10-12). G) Phosphorylation of IRE1 was measured in 3T3-L1 adipocytes after stimulation with vehicle, PGN (10 µg/mL), or thapsigargin (1 µM) for 3 hours, with or without 1 µM KirA6 (N=4-6). Blots were stripped and re-probed for total IRE1. H) Quantification of Western blots for the ratio between phosphorylated IRE1 and total IRE1, expressed relative to the basal control sample. I-K) RNA isolated from 3T3-L1 adipocytes was treated for 3 hours with vehicle, FK565 (PGN, 10 µg/mL), thapsigargin (1 µM), or isoproterenol (2 µM) in the absence or presence of KirA6 (1 µM). Expression of BiP (I), CHOP (J), and spliced Xbp1(K) (N=12). Values are mean ± SEM. Statistical significance was measured as p

    Techniques Used: Activity Assay, Expressing, Western Blot, Isolation

    35) Product Images from "Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems"

    Article Title: Automated Nucleic Acid Extraction Systems for Detecting Cytomegalovirus and Epstein-Barr Virus Using Real-Time PCR: A Comparison Study Between the QIAsymphony RGQ and QIAcube Systems

    Journal: Annals of Laboratory Medicine

    doi: 10.3343/alm.2017.37.2.129

    Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.
    Figure Legend Snippet: Discrepant cytomegalovirus (CMV, N=44; left panel) and Epstein-Barr virus (EBV, N=29; right panel) results using the QIAsymphony RGQ and QIAcube systems.

    Techniques Used:

    36) Product Images from "The Tonoplastic Inositol Transporter INT1 From Arabidopsis thaliana Impacts Cell Elongation in a Sucrose-Dependent Way"

    Article Title: The Tonoplastic Inositol Transporter INT1 From Arabidopsis thaliana Impacts Cell Elongation in a Sucrose-Dependent Way

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2018.01657

    qPCR analysis of genes involved in inositol metabolism in etiolated hypocotyls. Gene expression was analyzed in hypocotyls of 4-day-old etiolated WT and int1.1 seedlings grown on inositol-free medium without sucrose (=w/o) or with 2% sucrose added (=Suc). (A) Genes involved in inositol biosynthesis, inositol salvage pathway and cell wall biosynthesis. (B) Genes related to biosynthesis of inositol-containing membrane components and IP3-signaling. Relative gene expression levels were normalized to the reference gene ( UBI10 ). Re-adjustment was performed to the expression levels of WT hypocotyls grown on 0 mM sucrose, 0 mM inositol, which were set at 1.0. Mean values of three independent experiments are given; error bars = standard deviation. Differences between genotypes were analyzed using one-way ANOVA followed by Tukey HSD test. Black asterisks indicate a highly significant difference compared to WT on the respective medium, white asterisks indicate a highly significant difference between lines grown on medium without sucrose and the same plant line on medium with sucrose (one asterisk = p
    Figure Legend Snippet: qPCR analysis of genes involved in inositol metabolism in etiolated hypocotyls. Gene expression was analyzed in hypocotyls of 4-day-old etiolated WT and int1.1 seedlings grown on inositol-free medium without sucrose (=w/o) or with 2% sucrose added (=Suc). (A) Genes involved in inositol biosynthesis, inositol salvage pathway and cell wall biosynthesis. (B) Genes related to biosynthesis of inositol-containing membrane components and IP3-signaling. Relative gene expression levels were normalized to the reference gene ( UBI10 ). Re-adjustment was performed to the expression levels of WT hypocotyls grown on 0 mM sucrose, 0 mM inositol, which were set at 1.0. Mean values of three independent experiments are given; error bars = standard deviation. Differences between genotypes were analyzed using one-way ANOVA followed by Tukey HSD test. Black asterisks indicate a highly significant difference compared to WT on the respective medium, white asterisks indicate a highly significant difference between lines grown on medium without sucrose and the same plant line on medium with sucrose (one asterisk = p

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

    37) Product Images from "Detection of EGFR Mutations by TaqMan Mutation Detection Assays Powered by Competitive Allele-Specific TaqMan PCR Technology"

    Article Title: Detection of EGFR Mutations by TaqMan Mutation Detection Assays Powered by Competitive Allele-Specific TaqMan PCR Technology

    Journal: BioMed Research International

    doi: 10.1155/2013/385087

    Representative results for EGFR mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR TaqMan Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).
    Figure Legend Snippet: Representative results for EGFR mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR TaqMan Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).

    Techniques Used: Mutagenesis, Sequencing

    38) Product Images from "Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice"

    Article Title: Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19102959

    Expression of slow and fast muscle-associated genes in quadriceps muscles. ( A ) Myosin heavy chain gene expressions by qRT-PCR. Myh7 is represented as a slow muscle type, whereas Myh2 , Myh1 , and Myh4 are fast muscle types. ( B ) Comparisons of protein levels associated with slow muscle types between WT and TG in quadriceps muscle by western blotting. Overall, slow muscle type associated proteins were up-regulated in quadriceps muscles of TG mice. ( C ) Analysis of slow muscle-associated genes expression by qRT-PCR. ( D ) Analysis of fast muscle-associated genes expression by qRT-PCR. These data were from four-month-old wild-type ( n = 4) and transgenic mice ( n = 5), which were normalized according to the amount of mouse Gapdh mRNA and expressed relative to the corresponding value of the wild-type mice. Data are expressed as the means ± SEM for three independent replicates and multiple t -tests were used for the statistical analysis. * = p
    Figure Legend Snippet: Expression of slow and fast muscle-associated genes in quadriceps muscles. ( A ) Myosin heavy chain gene expressions by qRT-PCR. Myh7 is represented as a slow muscle type, whereas Myh2 , Myh1 , and Myh4 are fast muscle types. ( B ) Comparisons of protein levels associated with slow muscle types between WT and TG in quadriceps muscle by western blotting. Overall, slow muscle type associated proteins were up-regulated in quadriceps muscles of TG mice. ( C ) Analysis of slow muscle-associated genes expression by qRT-PCR. ( D ) Analysis of fast muscle-associated genes expression by qRT-PCR. These data were from four-month-old wild-type ( n = 4) and transgenic mice ( n = 5), which were normalized according to the amount of mouse Gapdh mRNA and expressed relative to the corresponding value of the wild-type mice. Data are expressed as the means ± SEM for three independent replicates and multiple t -tests were used for the statistical analysis. * = p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Mouse Assay, Transgenic Assay

    39) Product Images from "The Transcription Factor MEF2C Negatively Controls Angiogenic Sprouting of Endothelial Cells Depending on Oxygen"

    Article Title: The Transcription Factor MEF2C Negatively Controls Angiogenic Sprouting of Endothelial Cells Depending on Oxygen

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0101521

    A2M mediates the inhibitive effect of MEF2C on sprouting. (a) Inhibition of sprouting by purified A2M. Increasing concentrations of commercially available A2M were added to the spheroid sprouting assay and the effects on total sprout length scored. Data are displayed as mean values ± SEM as calculated from three experiments. (b) A2M mRNA expression is strongly reduced upon shRNA mediated knockdown. HUVEC were transduced with LV.shA2M or LV.shcon for 48 hours. Total RNA was isolated, subjected to cDNA synthesis and analyzed by realtime RT-PCR. Values were normalized to beta-2-microglobulin mRNA as internal standard and displayed as mean ±SD. One representative experiment of 3 performed in triplicates is shown (c) Knockdown of A2M restores sprouting activity in Ad.MEF2C transduced HUVEC. Cells were first transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 6 hours before they were used to form spheroids for the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was scored. Data displayed are mean values ± SEM calculated from three experiments. (d) Knockdown of A2M reduces the inhibitory activity in conditioned media from Ad.MEF2C transduced cells. HUVEC were transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 8 hours. Then medium was changed to serum-free Opti-MEM medium, supernatants were harvested after 48 hours, concentrated by diafiltration and added to the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was assessed. Values depicted are mean values ± SEM calculated from three independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. n.s. not significant, ***p
    Figure Legend Snippet: A2M mediates the inhibitive effect of MEF2C on sprouting. (a) Inhibition of sprouting by purified A2M. Increasing concentrations of commercially available A2M were added to the spheroid sprouting assay and the effects on total sprout length scored. Data are displayed as mean values ± SEM as calculated from three experiments. (b) A2M mRNA expression is strongly reduced upon shRNA mediated knockdown. HUVEC were transduced with LV.shA2M or LV.shcon for 48 hours. Total RNA was isolated, subjected to cDNA synthesis and analyzed by realtime RT-PCR. Values were normalized to beta-2-microglobulin mRNA as internal standard and displayed as mean ±SD. One representative experiment of 3 performed in triplicates is shown (c) Knockdown of A2M restores sprouting activity in Ad.MEF2C transduced HUVEC. Cells were first transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 6 hours before they were used to form spheroids for the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was scored. Data displayed are mean values ± SEM calculated from three experiments. (d) Knockdown of A2M reduces the inhibitory activity in conditioned media from Ad.MEF2C transduced cells. HUVEC were transduced with LV.shcon or LV.shA2M for 24 hours and then infected with Ad.con or Ad.MEF2C for 8 hours. Then medium was changed to serum-free Opti-MEM medium, supernatants were harvested after 48 hours, concentrated by diafiltration and added to the spheroid sprouting assay. Basal and VEGF-A/bFGF-induced sprouting was assessed. Values depicted are mean values ± SEM calculated from three independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. n.s. not significant, ***p

    Techniques Used: Inhibition, Purification, Expressing, shRNA, Transduction, Isolation, Reverse Transcription Polymerase Chain Reaction, Activity Assay, Infection, Diafiltration Assay

    Alpha-2-macroglobulin (A2M) is upregulated by MEF2C in HUVEC on mRNA and protein level. (a) A2M and MEF2C mRNA levels. HUVEC were transduced for 8, 16, 24 and 32 hours with Ad.MEF2C or Ad.con using MOI of 10 or left without infection (0 hours value). RNAs were isolated from the cells and A2M as well as MEF2C mRNA levels were determined by realtime RT-PCR as described in detail in the Methods section. Fold induction levels are shown as mean values ± SD calculated from triplicates of one representative experiment out of three independent experiments performed. Obtained values were normalized to beta-2-microglobulin mRNA as internal standard. (b) A2M protein. HUVEC were infected with Ad.MEF2C, Ad.con or left without infection for 8 hours, then medium was changed to serum-free Opti-MEM medium and supernatants were harvested after 48 hours. A2M secreted into supernatants was determined using a commercial A2M ELISA kit and a standard curve obtained with purified A2M. The mean concentration of A2M ± SD as calculated from three HUVEC batches analyzed is shown. (c) Supernatants of MEF2C virus transduced HUVEC inhibit sprouting. HUVEC were transduced with Ad.con or Ad.MEF2C or left uninduced for 24 hours, then medium was changed to Opti-mem, after 48 hours supernatants were collected and further concentrated by diafiltration as described in the Methods section. These conditioned media were added to the sprouting assay (50–70 µl to a 500 µl assay) and VEGF-A/bFGF-induced as well as basal sprout formation was scored . Data are displayed as mean values ± SEM as calculated from 3 independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. ***p
    Figure Legend Snippet: Alpha-2-macroglobulin (A2M) is upregulated by MEF2C in HUVEC on mRNA and protein level. (a) A2M and MEF2C mRNA levels. HUVEC were transduced for 8, 16, 24 and 32 hours with Ad.MEF2C or Ad.con using MOI of 10 or left without infection (0 hours value). RNAs were isolated from the cells and A2M as well as MEF2C mRNA levels were determined by realtime RT-PCR as described in detail in the Methods section. Fold induction levels are shown as mean values ± SD calculated from triplicates of one representative experiment out of three independent experiments performed. Obtained values were normalized to beta-2-microglobulin mRNA as internal standard. (b) A2M protein. HUVEC were infected with Ad.MEF2C, Ad.con or left without infection for 8 hours, then medium was changed to serum-free Opti-MEM medium and supernatants were harvested after 48 hours. A2M secreted into supernatants was determined using a commercial A2M ELISA kit and a standard curve obtained with purified A2M. The mean concentration of A2M ± SD as calculated from three HUVEC batches analyzed is shown. (c) Supernatants of MEF2C virus transduced HUVEC inhibit sprouting. HUVEC were transduced with Ad.con or Ad.MEF2C or left uninduced for 24 hours, then medium was changed to Opti-mem, after 48 hours supernatants were collected and further concentrated by diafiltration as described in the Methods section. These conditioned media were added to the sprouting assay (50–70 µl to a 500 µl assay) and VEGF-A/bFGF-induced as well as basal sprout formation was scored . Data are displayed as mean values ± SEM as calculated from 3 independent experiments. The values obtained for conditioned medium from Ad.con infected cells treated with VEGF and bFGF were arbitrarily set to 100%. ***p

    Techniques Used: Infection, Isolation, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Purification, Concentration Assay, Transduction, Diafiltration Assay

    40) Product Images from "Clonorchis sinensis omega-class glutathione transferases play major roles in the protection of the reproductive system during maturation and the response to oxidative stress"

    Article Title: Clonorchis sinensis omega-class glutathione transferases play major roles in the protection of the reproductive system during maturation and the response to oxidative stress

    Journal: Parasites & Vectors

    doi: 10.1186/s13071-016-1622-2

    Expression profiles of the C. sinensis GSTo1 and CsGSTo2 according to developmental stages. a Total RNAs (1 μg) extracted from each of the developmental stages were reverse transcribed as indicated on top. The mRNA transcripts of CsGSTo1 and 2 amplified by a semiquantitative RT-PCR were analyzed by 2 % agarose gels with ethidium bromide staining. C. sinensis tubulin gene ( CsTub ), which was shown to be constitutively expressed throughout developmental stages, was used as a control. The reaction mixture that did not contain reverse transcriptase during synthesis of the first single strand cDNA was used as a negative control. Abbreviations : Egg, C. sinensis egg; MC, metacercaria; 1 wk, 1-week-old juvenile; 2 wk, 2-week-old juvenile; 3 wk, 3-week-old immature; 4 wk, 4-week-old-mature C. sinensis . b Alteration of CsGSTo transcripts by qRT-PCR. The mRNA transcripts in each of the RNA samples (200 ng) were reverse-transcribed and the resulting cDNAs were employed in qRT-PCR as templates. The fold increase was calculated by differences in threshold cycles (ΔΔC T ) of the CsGSTo1 and 2 among different developmental stages. CsTub gene was used as normalization control. c Expressional changes of CsGSTo1 and 2 proteins determined by immunoblotting probed with anti-rCsGSTo1 and 2. C. sinensis tubulin (CsTub) was employed as a control. Each lane contained 100 ng protein
    Figure Legend Snippet: Expression profiles of the C. sinensis GSTo1 and CsGSTo2 according to developmental stages. a Total RNAs (1 μg) extracted from each of the developmental stages were reverse transcribed as indicated on top. The mRNA transcripts of CsGSTo1 and 2 amplified by a semiquantitative RT-PCR were analyzed by 2 % agarose gels with ethidium bromide staining. C. sinensis tubulin gene ( CsTub ), which was shown to be constitutively expressed throughout developmental stages, was used as a control. The reaction mixture that did not contain reverse transcriptase during synthesis of the first single strand cDNA was used as a negative control. Abbreviations : Egg, C. sinensis egg; MC, metacercaria; 1 wk, 1-week-old juvenile; 2 wk, 2-week-old juvenile; 3 wk, 3-week-old immature; 4 wk, 4-week-old-mature C. sinensis . b Alteration of CsGSTo transcripts by qRT-PCR. The mRNA transcripts in each of the RNA samples (200 ng) were reverse-transcribed and the resulting cDNAs were employed in qRT-PCR as templates. The fold increase was calculated by differences in threshold cycles (ΔΔC T ) of the CsGSTo1 and 2 among different developmental stages. CsTub gene was used as normalization control. c Expressional changes of CsGSTo1 and 2 proteins determined by immunoblotting probed with anti-rCsGSTo1 and 2. C. sinensis tubulin (CsTub) was employed as a control. Each lane contained 100 ng protein

    Techniques Used: Expressing, Amplification, Reverse Transcription Polymerase Chain Reaction, Staining, Negative Control, Quantitative RT-PCR

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    Qiagen rotor gene q real time pcr cycler
    Melting curve analysis of the targeted eight poxviruses using different <t>PCR</t> platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).
    Rotor Gene Q Real Time Pcr Cycler, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 180 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen rotor gene 6000 real time pcr cycler
    Melting curve analysis of the targeted eight poxviruses using different <t>PCR</t> platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).
    Rotor Gene 6000 Real Time Pcr Cycler, supplied by Qiagen, used in various techniques. Bioz Stars score: 91/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rotor gene 6000 real time pcr cycler - by Bioz Stars, 2020-09
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    91
    Qiagen real time pcr cycler
    Melting curve analysis of the targeted eight poxviruses using different <t>PCR</t> platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).
    Real Time Pcr Cycler, supplied by Qiagen, used in various techniques. Bioz Stars score: 91/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 51 article reviews
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    real time pcr cycler - by Bioz Stars, 2020-09
    91/100 stars
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    Melting curve analysis of the targeted eight poxviruses using different PCR platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).

    Journal: Scientific Reports

    Article Title: A novel HRM assay for the simultaneous detection and differentiation of eight poxviruses of medical and veterinary importance

    doi: 10.1038/srep42892

    Figure Lengend Snippet: Melting curve analysis of the targeted eight poxviruses using different PCR platforms. Each genotype displayed a unique melting peak. ( a ). amplification plot; ( b ). melt curve plot; ( c ). melting peaks using the QuantStudio 6, Life Technologies); ( d ). melting peaks using the CFX96, Bio-Rad; ( e ). melting peaks using LC480II, Roche; ( f ). melting peaks using the Rotor Gene Q, Qiagen); ( g ). Linearity test (10 9 to 10 2 virus copies); and ( h ). Co-infection with CMLV and camel PCPV (blue colour two melting peaks, 72.80 °C and 81.20 °C for CMLV and camel PCPV respectively).

    Article Snippet: Thus, using the same PCR mix and protocol as in the BioRad CFX96 instrument, PCR and melting curve acquisition analysis was also conducted on the LightCycler® 480 Real-Time PCR Systems (Roche), QuantStudio™ 6 Flex Real-Time PCR System (Life Technologies), and Rotor-Gene Q Real-Time PCR Cycler (Qiagen).

    Techniques: Polymerase Chain Reaction, Amplification, Infection