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  • 90
    Thermo Fisher sybr greener mirna qrt pcr kit
    <t>miRNA</t> alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. <t>qRT-PCR</t> analysis of the relative expression levels of miR-193b, miR-130b and miR-27a
    Sybr Greener Mirna Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transcript polymerase chain reaction qrt pcr kit
    <t>miRNA</t> alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. <t>qRT-PCR</t> analysis of the relative expression levels of miR-193b, miR-130b and miR-27a
    Transcript Polymerase Chain Reaction Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher transcription polymerase chain reaction
    <t>miRNA</t> alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. <t>qRT-PCR</t> analysis of the relative expression levels of miR-193b, miR-130b and miR-27a
    Transcription Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 561 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt polymerase chain reaction pcr
    <t>miRNA</t> alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. <t>qRT-PCR</t> analysis of the relative expression levels of miR-193b, miR-130b and miR-27a
    Qrt Polymerase Chain Reaction Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1001 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcriptase polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcription polymerase chain reaction qrt pcr
    Primer Design and <t>qRT-PCR</t>
    Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 540 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher time polymerase chain reaction
    Primer Design and <t>qRT-PCR</t>
    Time Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcribed polymerase chain reaction
    Primer Design and <t>qRT-PCR</t>
    Reverse Transcribed Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative reverse transcription polymerase chain reaction qrt pcr
    ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by <t>qRT-PCR</t> (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time polymerase chain reaction qrt pcr reagents
    <t>QRT-PCR</t> for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p
    Real Time Polymerase Chain Reaction Qrt Pcr Reagents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr trizol
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantificational reverse transcription polymerase chain reaction qrt pcr trizol
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Quantificational Reverse Transcription Polymerase Chain Reaction Qrt Pcr Trizol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher reverse transcriptase polymerase chain reaction
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Reverse Transcriptase Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time reverse transcription polymerase chain reaction
    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, <t>qRT‐PCR</t> result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P
    Real Time Reverse Transcription Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 558 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative reverse transcription polymerase chain reaction qrt pcr trizol reagent
    miR-628 suppressed the mRNA and protein expression of SOS1 . Notes: The breast CSCs of MDA-MB-231 ( A ) and MCF-7 ( B ) cells were transfected with the miR-628 mimic and miR-628 inhibitor, and SOS1 mRNA level was determined by <t>qRT-PCR.</t> ( C ) The breast CSCs of MDA-MB-231 and MCF-7 cells were transfected with miR-628 mimic and miR-628 inhibitor, and SOS1 protein level was analyzed by Western blotting. The protein expression levels were statistically analyzed by quantitating the intensity of the protein bands relative to that of the internal loading control (α-tubulin). Values indicate mean ± SD; * P
    Quantitative Reverse Transcription Polymerase Chain Reaction Qrt Pcr Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr trizol reagent
    Downregulation of SIRT4 expression correlates with clinicopathological factors and poor prognosis in GC patients. ( A ) The expression of SIRT4 in tumor tissues and adjacent normal tissues was determined by <t>qRT-PCR</t> and Western blotting, respectively. Tumor tissues vs normal adjacent tissues, * p
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 71 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 7900ht quantitative real time polymerase chain reaction qrt pcr system
    KMP 1 specifically recognized CD 44 epitope located on the cell surface in bladder cancer. (A) To investigate the epitope recognized by KMP 1, the supernatants after EJ cell lysis were applied to protein A‐Sepharose column with KMP 1. Two peaks were obtained from the eluates by Sephacryl S200. (B) Antigen–antibody complex peak and antibody excess peak obtained from the eluates were resolved by 10% SDS – PAGE and stained with silver. (C) <t>qRT</t> – <t>PCR</t> analysis of CD 44 expression in EJ cells that were transfected with pS uper‐puro‐ CD 44‐sh RNA (sh CD 44‐ EJ ) or the empty pS uper‐puro vector (shCtrl‐ EJ ). (D) Western blot imagines indicated that the band using KMP 1 as a primary antibody was markedly thinned in the CD 44‐silenced EJ cells versus the shCtrl‐ EJ cells. (E) sh CD 44‐ EJ and shCtrl‐ EJ cells were analyzed by flow cytometry after staining with KMP 1, and nontransfected EJ cells were stained with mIgG as a negative control. (F) ELISA analysis of KMP 1 recognized CD 44 levels in shCtrl or sh CD 44‐ EJ cells. Each bar is expressed as the mean ± standard deviation of three experiments. * P
    7900ht Quantitative Real Time Polymerase Chain Reaction Qrt Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii one step qrt polymerase chain reaction pcr system
    KMP 1 specifically recognized CD 44 epitope located on the cell surface in bladder cancer. (A) To investigate the epitope recognized by KMP 1, the supernatants after EJ cell lysis were applied to protein A‐Sepharose column with KMP 1. Two peaks were obtained from the eluates by Sephacryl S200. (B) Antigen–antibody complex peak and antibody excess peak obtained from the eluates were resolved by 10% SDS – PAGE and stained with silver. (C) <t>qRT</t> – <t>PCR</t> analysis of CD 44 expression in EJ cells that were transfected with pS uper‐puro‐ CD 44‐sh RNA (sh CD 44‐ EJ ) or the empty pS uper‐puro vector (shCtrl‐ EJ ). (D) Western blot imagines indicated that the band using KMP 1 as a primary antibody was markedly thinned in the CD 44‐silenced EJ cells versus the shCtrl‐ EJ cells. (E) sh CD 44‐ EJ and shCtrl‐ EJ cells were analyzed by flow cytometry after staining with KMP 1, and nontransfected EJ cells were stained with mIgG as a negative control. (F) ELISA analysis of KMP 1 recognized CD 44 levels in shCtrl or sh CD 44‐ EJ cells. Each bar is expressed as the mean ± standard deviation of three experiments. * P
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    Thermo Fisher quantitative reverse transcriptase polymerase chain reaction qrt pcr trizol reagent
    Expression of proliferation and apoptosis associated genes and proteins. a mRNA expression of Akt, Erk, Casp3 and Casp9 using <t>qRT-PCR;</t> b protein expression of p-Akt, p-Erk, pho-Casp3 and pho-Casp9; c gray value of p-Akt, p-Erk, pho-Casp3 and pho-Casp9 using Western blotting. * P
    Quantitative Reverse Transcriptase Polymerase Chain Reaction Qrt Pcr Trizol Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression of proliferation and apoptosis associated genes and proteins. a mRNA expression of Akt, Erk, Casp3 and Casp9 using <t>qRT-PCR;</t> b protein expression of p-Akt, p-Erk, pho-Casp3 and pho-Casp9; c gray value of p-Akt, p-Erk, pho-Casp3 and pho-Casp9 using Western blotting. * P
    Real Time Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression of proliferation and apoptosis associated genes and proteins. a mRNA expression of Akt, Erk, Casp3 and Casp9 using <t>qRT-PCR;</t> b protein expression of p-Akt, p-Erk, pho-Casp3 and pho-Casp9; c gray value of p-Akt, p-Erk, pho-Casp3 and pho-Casp9 using Western blotting. * P
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    HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by <t>qRT‐PCR.</t> Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P
    Powersybr Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Kits, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher stepone real time quantitative polymerase chain reaction qrt pcr
    HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by <t>qRT‐PCR.</t> Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P
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    GATA3 inhibits epithelial–mesenchymal transition (EMT) in gastric cancer cells. Notes: ( A ) CTC-141 cells were transfected with vector, GATA3 and GATA3 (R298Q), and the relative expressions of EMT-associated genes were detected by <t>qRT-PCR.</t> * P
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    Image Search Results


    miRNA alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. qRT-PCR analysis of the relative expression levels of miR-193b, miR-130b and miR-27a

    Journal: Chinese Journal of Cancer Research

    Article Title: TGF-?1 alters microRNA profile in human gastric cancer cells

    doi: 10.3978/j.issn.1000-9604.2013.01.09

    Figure Lengend Snippet: miRNA alternation induced by TGF-β1 treatment (n=3). A. The miRNA microarray clustering results of BGC823/0 h, BGC823/24 hrs and BGC823/36 hrs cell lines; B. qRT-PCR analysis of the relative expression levels of miR-193b, miR-130b and miR-27a

    Article Snippet: Real-time PCR was performed using express SYBR GreenER miRNA qRT-PCR Kit (Invitrogen) on 7,500 real-time PCR system (ABI, USA).

    Techniques: Microarray, Quantitative RT-PCR, Expressing

    Primer Design and qRT-PCR

    Journal: Plastic and reconstructive surgery

    Article Title: Beta-catenin-dependent Wnt signaling: a pathway in acute cutaneous wounding

    doi: 10.1097/PRS.0000000000004170

    Figure Lengend Snippet: Primer Design and qRT-PCR

    Article Snippet: All other reagents for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Applied Biosystems (Foster City, CA). rmWnt3a was from R & D Systems (Minneapolis, MN).

    Techniques: Quantitative RT-PCR

    Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Bleomycin induces collagen deposition in the lungs and reduces survival in Myr-Akt mice. Myr-Akt and wild type (WT) littermate mice were treated twice a week with bleomycin (0.035 U/g) or PBS (vehicle) for 4 weeks, as described in Material and Methods. (a) Mouse survival was recorded and plotted using a Kaplan-Meier curve. Myr-Akt mice had worse survival than WT mice in response to bleomycin treatment. N = 7 mice for each WT and Myr-Akt mouse group. (b) All surviving mice were sacrificed at day 33 and the lungs formalin-fixed, sectioned, and stained with Masson’s Trichrome. Five images per slide were captured. Red staining represents keratin, light red and pink staining represents cytoplasm, dark brown represents cell nuclei, and blue staining represents collagen. The images are representative images from 3 mice per group using a 20X objective (200× magnification). (c) The images were quantified blindly by histogram analysis using Adobe Photoshop CS and the percent of blue staining per high-power field (HPF) from ten images and expressed as mean ± SEM. (d) Lungs from WT and Myr-Akt mice subjected to PBS or bleomycin stained with H E, Trichrome, and F4/80 were evaluated by a board-certified pathologist for inflammation score (0 to 5 point scale). (e) Total RNA was extracted from homogenized lung tissue using TRIzol. cDNA was synthesized and qRT-PCR performed for mouse collagen IA and IIIB. Data are expressed as mean relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Mouse Assay, Staining, Synthesized, Quantitative RT-PCR, IA, Expressing

    Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Reduced autophagy in the lungs of Myr-Akt mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to LC3B immunohistochemical staining. Ten pictures per section were imaged using 10X and 40X objectives (100× and 400× magnification, respectively). (b) Images were quantified using histogram analysis feature in Adobe Photoshop CS5 software for percent brown stain (F4/80+ pixels) per high-power field (HPF) from at least five images per section and expressed as mean ± SEM. (c) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse Beclin-1 , Lc 3 a , and Lc 3 b mRNAs. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (d) Total protein were isolated and equal amounts of protein separated on a 4% – 12% SDS-PAGE gel and immunoblotted for BECLIN-1, LC3A/B-I and LC3A/B-II, and β -ACTIN. (e) Total RNA was extracted from homogenized lungs using TRIzol. cDNA was synthesized and qRT-PCR performed using primers specific for mouse p 62/ SQSTM1 mRNA. Data are expressed as relative copy number (RCN) of mRNA expression over average endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represents the mean RCN ± SEM. (f) Immunoblot for phosphor-mTOR (ser2448) and mTOR (Cell Signaling Technology). Bands were quantified using densitometry by comparing band density over β -ACTIN or total mTOR in each respective lane. Data are expressed as the mean densitometry value ± SEM. 4 pairs of age-matched littermate mice were used for western analysis and 5 pairs of age-matched littermate mice were used for gene expression.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Software, Synthesized, Quantitative RT-PCR, Expressing, Isolation, SDS Page, Western Blot

    Differential myofibroblast expression in the lungs of Myr-Akt and WT mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to α -SMA immunohistochemical staining for myofibroblasts. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified by histogram analysis using Adobe Photoshop CS5 software and percent brown stain ( α -SMA+ pixels) per high-power field (HPF) from at least five images per section are expressed as mean ± SEM. (c) Total RNA was extracted from Myr-Akt or WT littermate mouse lungs and α -SMA mRNA was measured by qRT-PCR. Data are expressed as relative copy number (RCN) of α -SMA mRNA expression over the average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represent the mean RCN ± SEM. Single-cell suspension from lung homogenate were immunostained using mouse α-SMA antibodies or double-immunostained with α -SMA and M-CSF-R (CD115) antibodies and analyzed by flow cytometry. (d) Data represents mean percent of α -SMA+ cells and (e) α -SMA+/M-CSF-R+ cells ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Differential myofibroblast expression in the lungs of Myr-Akt and WT mice. (a) 6 – 8 week old Myr-Akt or WT mice were sacrificed and lungs harvested, paraffin embedded, and subjected to α -SMA immunohistochemical staining for myofibroblasts. Ten pictures per section were imaged using 4X and 20X objectives (40× and 200× magnification, respectively). (b) Images were quantified by histogram analysis using Adobe Photoshop CS5 software and percent brown stain ( α -SMA+ pixels) per high-power field (HPF) from at least five images per section are expressed as mean ± SEM. (c) Total RNA was extracted from Myr-Akt or WT littermate mouse lungs and α -SMA mRNA was measured by qRT-PCR. Data are expressed as relative copy number (RCN) of α -SMA mRNA expression over the average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh ). Data represent the mean RCN ± SEM. Single-cell suspension from lung homogenate were immunostained using mouse α-SMA antibodies or double-immunostained with α -SMA and M-CSF-R (CD115) antibodies and analyzed by flow cytometry. (d) Data represents mean percent of α -SMA+ cells and (e) α -SMA+/M-CSF-R+ cells ± SEM. 5 pairs of age-matched littermate mice were used for each experiment.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Expressing, Mouse Assay, Immunohistochemistry, Staining, Software, Quantitative RT-PCR, Flow Cytometry, Cytometry

    Bleomycin suppresses autophagy gene expression in the lungs of Myr-Akt mice. Total RNA was extracted from paraffin embedded lung tissue section with RecoverAll Total Nucleic Acid Isolation Kit for FFPE according to manufacturer’s protocol. cDNA was synthesized and qRT-PCR performed for mouse Beclin- 1, Lc 3 a , and Lc 3 b . Data are expressed as relative copy number (RCN) of mRNA expression over average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh .). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Journal: Advances in bioscience and biotechnology (Print)

    Article Title: Constitutive AKT Activity Predisposes Lung Fibrosis by Regulating Macrophage, Myofibroblast and Fibrocyte Recruitment and Changes in Autophagy

    doi: 10.4236/abb.2019.1010027

    Figure Lengend Snippet: Bleomycin suppresses autophagy gene expression in the lungs of Myr-Akt mice. Total RNA was extracted from paraffin embedded lung tissue section with RecoverAll Total Nucleic Acid Isolation Kit for FFPE according to manufacturer’s protocol. cDNA was synthesized and qRT-PCR performed for mouse Beclin- 1, Lc 3 a , and Lc 3 b . Data are expressed as relative copy number (RCN) of mRNA expression over average of endogenous control RNAs ( Cap 1, Rpl 4 and Gapdh .). Data represents the mean RCN ± SEM and N = 4 mice per group.

    Article Snippet: Primers and SYBR Green Master Mix (4385610) for quantitative real-time polymerase chain reaction (qRT-PCR) were purchased from Thermofisher Scientific unless indicated otherwise.

    Techniques: Expressing, Mouse Assay, Isolation, Formalin-fixed Paraffin-Embedded, Synthesized, Quantitative RT-PCR

    ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by qRT-PCR (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.

    Journal: International Journal of Molecular Sciences

    Article Title: MicroRNA Expression Profile of Neural Progenitor-Like Cells Derived from Rat Bone Marrow Mesenchymal Stem Cells under the Influence of IGF-1, bFGF and EGF

    doi: 10.3390/ijms16059693

    Figure Lengend Snippet: ( A ) Hierarchal clustering of differentially expressed microRNAs. Forty-six differentially expressed microRNAs (for up- and down-regulation, respectively) compared to the negative control group in at least one sample were clustered hierarchically. Each rows represent individual microRNAs. The expression ratio representing colour ranges from green (low) to red (high), as indicated by the scale bar. The column bar chart shows the relative expression level of selected microRNAs by qRT-PCR (right column, white) as compared to the fold changes of the corresponding microRNA (left column, grey); ( B ) MicroRNAs expression of Group B versus Group A on the first day of treatment; ( C ) Group B versus Group A on the third day of treatment; ( D ) MicroRNAs expression of on the fifth day of treatment. U87 was used as housekeeping gene. Bars represent the mean ± standard error of three independent experiments; and ( E ) qRT-PCR product was confirmed by gel electrophoresis at 90 V for 40 min.

    Article Snippet: MicroRNA-qPCR Assay MicroRNA expressions were validated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) according to the manufacturer’s protocol (all from Applied Biosystems, Life Technologies Co.; Waltham, MA, USA).

    Techniques: Negative Control, Expressing, Quantitative RT-PCR, Nucleic Acid Electrophoresis

    QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 cells exposed to prostaglandins. (a) 27-Hydroxylase message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 1 , E 2 , and D 2 . THP-1 human monocytes were exposed to the following conditions represented by the six bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 1 (0.1 μM) + NS398 (50 μM), (4) PGE 2 (0.1 μM) + NS398 (50 μM), (5) PGE 1 (0.1 μM) + PGE 2 (0.1 μM) + NS398 (50 μM), and (6) PGD 2 (14 μM) + NS398 (50 μM) (all 18-hour exposures). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. ** p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Amplification

    QRT-PCR for 27-hydroxylase and ABCA1 message in indomethacin-treated THP-1 cells. (a) 27-Hydroxylase mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. At 50 mM indomethacin concentration, cell death was statistically significant (16.8% ± 1.0%). ABCA1, ATP-binding cassette transporter A1; COX, cyclooxygenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; QRT-PCR, quantitative real-time polymerase chain reaction. * p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in indomethacin-treated THP-1 cells. (a) 27-Hydroxylase mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 mRNA expression is decreased by the non-specific COX inhibitor indomethacin in a dose-dependent fashion in THP-1 monocytes. Cultured THP-1 monocytic cells were untreated or exposed to increasing doses of indomethacin for 18 hours. After isolation of total RNA, the RNA was reverse-transcribed and the cDNA amplified by QRT-PCR as described. Signals obtained from the amplification of GAPDH message were used as internal controls. At 50 mM indomethacin concentration, cell death was statistically significant (16.8% ± 1.0%). ABCA1, ATP-binding cassette transporter A1; COX, cyclooxygenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; QRT-PCR, quantitative real-time polymerase chain reaction. * p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Isolation, Amplification, Concentration Assay, Binding Assay, Real-time Polymerase Chain Reaction

    QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 macrophages exposed to prostaglandins or TXA 2 . (a) 27-Hydroxylase message in THP-1 macrophages is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 in THP-1 macrophages and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. * p

    Journal: Arthritis Research & Therapy

    Article Title: Effect of cyclooxygenase inhibition on cholesterol efflux proteins and atheromatous foam cell transformation in THP-1 human macrophages: a possible mechanism for increased cardiovascular risk

    doi: 10.1186/ar2109

    Figure Lengend Snippet: QRT-PCR for 27-hydroxylase and ABCA1 message in NS398-treated THP-1 macrophages exposed to prostaglandins or TXA 2 . (a) 27-Hydroxylase message in THP-1 macrophages is decreased by the COX-2 inhibitor NS398 and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for 27-hydroxylase mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. (b) ABCA1 message is decreased by the COX-2 inhibitor NS398 in THP-1 macrophages and this decrease is reversed by prostaglandins E 2 and D 2 , but not TXA 2 . THP-1 human macrophages were exposed to the following conditions represented by the five bars (from left to right): (1) RPMI 1640, (2) NS398 (50 μM), (3) PGE 2 (0.1 μM) + NS398 (50 μM), (4) PGD 2 (14 μM) + NS398 (50 μM), and (5) TXA 2 (3 μM) + NS398 (50 μM) (24-hour exposures to NS398 alone followed by addition of indicated PG or TXA 2 for a further 24 hours). Cells were extracted for total RNA and were evaluated for ABCA1 mRNA expression by QRT-PCR. Signals obtained from the amplification of GAPDH message were used as internal controls. * p

    Article Snippet: All reagents for reverse transcription and quantitative real-time polymerase chain reaction (QRT-PCR) were purchased from Applied Biosystems (Foster City, CA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Amplification

    ASCs express the machinery for production and secretion of ACh. (A) Real‐time qRT‐PCR comparison of the expression of Chat and Slc18a3 . Compared with uASCs, the dASCs had a significantly higher expression of Chat and Slc18a3 . Slc18a3, vesicular acetylcholine transporter. (B) Immunocytochemistry of uASCs and dASCs using antibody against choline acetyltransferase (ChAT; red). DAPI was used to stain the nuclei (blue). * P

    Journal: Muscle & Nerve

    Article Title: Adipose stem cells enhance myoblast proliferation via acetylcholine and extracellular signal–regulated kinase 1/2 signaling

    doi: 10.1002/mus.25741

    Figure Lengend Snippet: ASCs express the machinery for production and secretion of ACh. (A) Real‐time qRT‐PCR comparison of the expression of Chat and Slc18a3 . Compared with uASCs, the dASCs had a significantly higher expression of Chat and Slc18a3 . Slc18a3, vesicular acetylcholine transporter. (B) Immunocytochemistry of uASCs and dASCs using antibody against choline acetyltransferase (ChAT; red). DAPI was used to stain the nuclei (blue). * P

    Article Snippet: Real‐time quantitative reverse transcript polymerase chain reaction (qRT‐PCR) was performed with a gene expression assay (TaqMan; Applied Biosystems) and real‐time PCR system (ViiA 7; Applied Biosystems).

    Techniques: Quantitative RT-PCR, Expressing, Immunocytochemistry, Staining

    CTHRC1 is frequently up-regulated in human breast cancer cells and tissues. a and b CTHRC1 was up-regulated in human breast cancer cells analyzed by qRT-PCR and western blot. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Collagen triple helix repeat containing-1 negatively regulated by microRNA-30c promotes cell proliferation and metastasis and indicates poor prognosis in breast cancer

    doi: 10.1186/s13046-017-0564-7

    Figure Lengend Snippet: CTHRC1 is frequently up-regulated in human breast cancer cells and tissues. a and b CTHRC1 was up-regulated in human breast cancer cells analyzed by qRT-PCR and western blot. * P

    Article Snippet: RNA extraction and real-time quantitative polymerase chain reaction (qRT-PCR) TRIzol® Reagent (Life Technologies, Carlsbad, CA) was used to isolate total RNA from frozen patient samples and cell lines according to the manufacturer’s protocol. cDNA was synthesized using the universal cDNA synthesis kit (Toyobo, tokyo, JP).

    Techniques: Quantitative RT-PCR, Western Blot

    CTHRC1 is a direct target of miR-30c. a qRT-PCR analysis of CTHRC1 mRNA expression in indicated cells 24 h post-transfection. ** P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Collagen triple helix repeat containing-1 negatively regulated by microRNA-30c promotes cell proliferation and metastasis and indicates poor prognosis in breast cancer

    doi: 10.1186/s13046-017-0564-7

    Figure Lengend Snippet: CTHRC1 is a direct target of miR-30c. a qRT-PCR analysis of CTHRC1 mRNA expression in indicated cells 24 h post-transfection. ** P

    Article Snippet: RNA extraction and real-time quantitative polymerase chain reaction (qRT-PCR) TRIzol® Reagent (Life Technologies, Carlsbad, CA) was used to isolate total RNA from frozen patient samples and cell lines according to the manufacturer’s protocol. cDNA was synthesized using the universal cDNA synthesis kit (Toyobo, tokyo, JP).

    Techniques: Quantitative RT-PCR, Expressing, Transfection

    CTHRC1 and miR-30c expression are inversely correlated in human breast cancer cells and tissues. a The relative expression level of miR-134, miR-155, miR-30c and miR-630 in breast cancer cells respectively was detected by qRT-PCR. * P

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: Collagen triple helix repeat containing-1 negatively regulated by microRNA-30c promotes cell proliferation and metastasis and indicates poor prognosis in breast cancer

    doi: 10.1186/s13046-017-0564-7

    Figure Lengend Snippet: CTHRC1 and miR-30c expression are inversely correlated in human breast cancer cells and tissues. a The relative expression level of miR-134, miR-155, miR-30c and miR-630 in breast cancer cells respectively was detected by qRT-PCR. * P

    Article Snippet: RNA extraction and real-time quantitative polymerase chain reaction (qRT-PCR) TRIzol® Reagent (Life Technologies, Carlsbad, CA) was used to isolate total RNA from frozen patient samples and cell lines according to the manufacturer’s protocol. cDNA was synthesized using the universal cDNA synthesis kit (Toyobo, tokyo, JP).

    Techniques: Expressing, Quantitative RT-PCR

    Effects of NRF2 and BACH1 on the induction of HO-1 by cigarette smoke. HBE1 cells were transfected with control non-specific or gene-specific siRNA. After 48 h of transfection, these cells were exposed to smoke extract for additional 24 h and then subjected to qRT-PCR. ( A , B ) Cells were transfected with siRNA targeting NRF2 and BACH1. The knockdown efficiency of NRF2 siRNA assessed by qRT-PCR was 81% in air exposed cells and 92% in smoke exposed cells ( A ); in BACH1 siRNA treatment, the knockdown efficiency was 81% in air exposed cells and 79% in smoke exposed cells ( B ); smoke had no effect on siRNA knockdown. ( C , D ) Cells with siRNA knockdown of NRF2 and BACH1 followed by smoke exposure were subjected to qRT-PCR for HO-1 ( C ) and CYP1A1 ( D ) expression. Results are expressed as the mean across three independent experiments with error bars representing standard errors of measurements (S.E.M.). * p

    Journal: International Journal of Molecular Sciences

    Article Title: Cigarette Smoke Regulates the Competitive Interactions between NRF2 and BACH1 for Heme Oxygenase-1 Induction

    doi: 10.3390/ijms18112386

    Figure Lengend Snippet: Effects of NRF2 and BACH1 on the induction of HO-1 by cigarette smoke. HBE1 cells were transfected with control non-specific or gene-specific siRNA. After 48 h of transfection, these cells were exposed to smoke extract for additional 24 h and then subjected to qRT-PCR. ( A , B ) Cells were transfected with siRNA targeting NRF2 and BACH1. The knockdown efficiency of NRF2 siRNA assessed by qRT-PCR was 81% in air exposed cells and 92% in smoke exposed cells ( A ); in BACH1 siRNA treatment, the knockdown efficiency was 81% in air exposed cells and 79% in smoke exposed cells ( B ); smoke had no effect on siRNA knockdown. ( C , D ) Cells with siRNA knockdown of NRF2 and BACH1 followed by smoke exposure were subjected to qRT-PCR for HO-1 ( C ) and CYP1A1 ( D ) expression. Results are expressed as the mean across three independent experiments with error bars representing standard errors of measurements (S.E.M.). * p

    Article Snippet: Real-Time Quantitative Reverse Transcriptase Polymerase Chain Reaction Quantitative RT-PCR (qRT-PCR) was carried out on an Applied Biosystems (Foster City, CA, USA) 5700 thermocycler as described previously [ ].

    Techniques: Transfection, Quantitative RT-PCR, Expressing

    Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, qRT‐PCR result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: Inhibition of LINC01116 suppressed cell proliferation and migration in NPC cells. A, qRT‐PCR result of LINC01116 expression in NPC cell lines and the normal HNEpC cells. B, qRT‐PCR result of LINC01116 expression in CNE2 and 5‐8F cells under the transfection of shCtrl or two shRNAs against LINC01116. C, CCK‐8 assay was performed to evaluate cell viability in above cells. D, Cell proliferation was examined by EdU assay. E, Transwell assay for the assessment of cell migration in NPC cells with or without LINC01116 inhibition. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Inhibition, Migration, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, EdU Assay, Transwell Assay

    MYC activated LINC01116 at transcriptional level in NPC. A‐D, The expression of MYC or LINC01116 in indicated CNE2 and 5‐8F cells was tested by qRT‐PCR. E, ChIP assay and AGE analysis verified the binding of MYC protein to LINC01116 promoter. F,G, Luciferase reporter assay confirmed that LINC01116 was transcriptionally activated by MYC in both CNE2 and 5‐8F cells. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: MYC activated LINC01116 at transcriptional level in NPC. A‐D, The expression of MYC or LINC01116 in indicated CNE2 and 5‐8F cells was tested by qRT‐PCR. E, ChIP assay and AGE analysis verified the binding of MYC protein to LINC01116 promoter. F,G, Luciferase reporter assay confirmed that LINC01116 was transcriptionally activated by MYC in both CNE2 and 5‐8F cells. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Binding Assay, Luciferase, Reporter Assay

    LINC01116 mainly located in the cytoplasm of NPC cells and interacted with the 5ʹUTR of MYC mRNA. A, LINC01116 was predicted by lncLocator as a cytoplasmic lncRNA. B, Subcellular fractionation plus qRT‐PCR validated that LINC01116 was mainly distributed in the cytoplasm of NPC cells. C, The online IntaRNA tool suggested that LINC01116 could bind to the 5'UTR of MYC mRNA. D, The interaction between LINC01116 and MYC mRNA in CNE2 and 5‐8F cells was confirmed by RNA pull down assay. ** P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: LINC01116 mainly located in the cytoplasm of NPC cells and interacted with the 5ʹUTR of MYC mRNA. A, LINC01116 was predicted by lncLocator as a cytoplasmic lncRNA. B, Subcellular fractionation plus qRT‐PCR validated that LINC01116 was mainly distributed in the cytoplasm of NPC cells. C, The online IntaRNA tool suggested that LINC01116 could bind to the 5'UTR of MYC mRNA. D, The interaction between LINC01116 and MYC mRNA in CNE2 and 5‐8F cells was confirmed by RNA pull down assay. ** P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Fractionation, Quantitative RT-PCR, Pull Down Assay

    LINC01116 enhanced MYC protein level so as to promote the transcription of MYC targets. A,B, The impact of LINC01116 on the mRNA and protein levels of MYC was estimated by qRT‐PCR (A) and western blot (B). C, Changes on the transcription of three MYC targets (CDK4, CCND2, and BMI1) in the context of MYC overexpression or together with LINC01116 suppression were evaluated by luciferase reporter assays. D‐E, qRT‐PCR results of the expression of CDK4, CCND2, and BMI1 in LINC01116‐silenced NPC cells. F, The influence of LINC01116 per se on the transcription of CDK4, CCND2, and BMI1 in NPC cells was determined by luciferase reporter assay. * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: LINC01116 enhanced MYC protein level so as to promote the transcription of MYC targets. A,B, The impact of LINC01116 on the mRNA and protein levels of MYC was estimated by qRT‐PCR (A) and western blot (B). C, Changes on the transcription of three MYC targets (CDK4, CCND2, and BMI1) in the context of MYC overexpression or together with LINC01116 suppression were evaluated by luciferase reporter assays. D‐E, qRT‐PCR results of the expression of CDK4, CCND2, and BMI1 in LINC01116‐silenced NPC cells. F, The influence of LINC01116 per se on the transcription of CDK4, CCND2, and BMI1 in NPC cells was determined by luciferase reporter assay. * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Quantitative RT-PCR, Western Blot, Over Expression, Luciferase, Expressing, Reporter Assay

    Overexpression of MYC rescued LINC01116 depletion‐restrained proliferation and migration in NPC cells. A, The expression of MYC at both mRNA and protein levels in indicated cells was detected via qRT‐PCR and western blot. B‐D, The viability, proliferation, and migration in 5‐8F cells under different conditions were respectively evaluated through CCK‐8 (B), EdU (C), and transwell assays (D). * P

    Journal: Cancer Medicine

    Article Title: LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity. LINC01116 accelerates nasopharyngeal carcinoma progression based on its enhancement on MYC transcription activity

    doi: 10.1002/cam4.2624

    Figure Lengend Snippet: Overexpression of MYC rescued LINC01116 depletion‐restrained proliferation and migration in NPC cells. A, The expression of MYC at both mRNA and protein levels in indicated cells was detected via qRT‐PCR and western blot. B‐D, The viability, proliferation, and migration in 5‐8F cells under different conditions were respectively evaluated through CCK‐8 (B), EdU (C), and transwell assays (D). * P

    Article Snippet: 2.3 Quantitative real‐time polymerase chain reaction (QRT‐PCR) TRIzol reagent from Invitrogen was utilized to isolate total RNAs from cultured NPC cells.

    Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay

    miR-628 suppressed the mRNA and protein expression of SOS1 . Notes: The breast CSCs of MDA-MB-231 ( A ) and MCF-7 ( B ) cells were transfected with the miR-628 mimic and miR-628 inhibitor, and SOS1 mRNA level was determined by qRT-PCR. ( C ) The breast CSCs of MDA-MB-231 and MCF-7 cells were transfected with miR-628 mimic and miR-628 inhibitor, and SOS1 protein level was analyzed by Western blotting. The protein expression levels were statistically analyzed by quantitating the intensity of the protein bands relative to that of the internal loading control (α-tubulin). Values indicate mean ± SD; * P

    Journal: OncoTargets and therapy

    Article Title: MicroRNA 628 suppresses migration and invasion of breast cancer stem cells through targeting SOS1

    doi: 10.2147/OTT.S164575

    Figure Lengend Snippet: miR-628 suppressed the mRNA and protein expression of SOS1 . Notes: The breast CSCs of MDA-MB-231 ( A ) and MCF-7 ( B ) cells were transfected with the miR-628 mimic and miR-628 inhibitor, and SOS1 mRNA level was determined by qRT-PCR. ( C ) The breast CSCs of MDA-MB-231 and MCF-7 cells were transfected with miR-628 mimic and miR-628 inhibitor, and SOS1 protein level was analyzed by Western blotting. The protein expression levels were statistically analyzed by quantitating the intensity of the protein bands relative to that of the internal loading control (α-tubulin). Values indicate mean ± SD; * P

    Article Snippet: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was used to extract total RNA following the manufacturer’s instructions.

    Techniques: Expressing, Multiple Displacement Amplification, Transfection, Quantitative RT-PCR, Western Blot

    Effect of miR-628 overexpression on migration and invasion of the breast CSC subpopulation in MCF-7 and MDA-MB-231 cell lines. Notes: ( A ) miR-628 expression after miR-628 mimic was transfected into CSCs of MCF-7 and MDA-MB-231 cells was determined by qRT-PCR. ( B – D ) Effects of miR-628 overexpression on migration ( B , C ) and invasion ( D , E ) of MCF-7 and MDA-MB-231 CSCs. ( F ) Cells were transfected with the miR-628 mimic. The effect of miR-628 mimic transfection on the protein levels of vimentin, Snail, and E-cadherin was analyzed by Western blotting. ( G ) Protein expression levels were statistically analyzed by quantitating the intensity of the protein bands relative to that of the internal loading control (α-tubulin). *Values indicate mean ± SD; P

    Journal: OncoTargets and therapy

    Article Title: MicroRNA 628 suppresses migration and invasion of breast cancer stem cells through targeting SOS1

    doi: 10.2147/OTT.S164575

    Figure Lengend Snippet: Effect of miR-628 overexpression on migration and invasion of the breast CSC subpopulation in MCF-7 and MDA-MB-231 cell lines. Notes: ( A ) miR-628 expression after miR-628 mimic was transfected into CSCs of MCF-7 and MDA-MB-231 cells was determined by qRT-PCR. ( B – D ) Effects of miR-628 overexpression on migration ( B , C ) and invasion ( D , E ) of MCF-7 and MDA-MB-231 CSCs. ( F ) Cells were transfected with the miR-628 mimic. The effect of miR-628 mimic transfection on the protein levels of vimentin, Snail, and E-cadherin was analyzed by Western blotting. ( G ) Protein expression levels were statistically analyzed by quantitating the intensity of the protein bands relative to that of the internal loading control (α-tubulin). *Values indicate mean ± SD; P

    Article Snippet: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) TRIzol reagent (Invitrogen, Carlsbad, CA, USA) was used to extract total RNA following the manufacturer’s instructions.

    Techniques: Over Expression, Migration, Multiple Displacement Amplification, Expressing, Transfection, Quantitative RT-PCR, Western Blot

    Downregulation of SIRT4 expression correlates with clinicopathological factors and poor prognosis in GC patients. ( A ) The expression of SIRT4 in tumor tissues and adjacent normal tissues was determined by qRT-PCR and Western blotting, respectively. Tumor tissues vs normal adjacent tissues, * p

    Journal: OncoTargets and therapy

    Article Title: SIRT4 acts as a tumor suppressor in gastric cancer by inhibiting cell proliferation, migration, and invasion

    doi: 10.2147/OTT.S156143

    Figure Lengend Snippet: Downregulation of SIRT4 expression correlates with clinicopathological factors and poor prognosis in GC patients. ( A ) The expression of SIRT4 in tumor tissues and adjacent normal tissues was determined by qRT-PCR and Western blotting, respectively. Tumor tissues vs normal adjacent tissues, * p

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) TRIzol reagent (Invitrogen) was used to extract total RNA from tissues or cells according to the manufacturer’s instructions.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    SIRT4 suppresses EMT through promoting E-cadherin expression. ( A ) SIRT4 was overexpressed or knocked down in MKN-45 or HGC-27 cells. After transfection for 48 h, the mRNA level of E-cadherin was detected using qRT-PCR. SIRT4 vs vector and siSIRT4 vs SCR, * p

    Journal: OncoTargets and therapy

    Article Title: SIRT4 acts as a tumor suppressor in gastric cancer by inhibiting cell proliferation, migration, and invasion

    doi: 10.2147/OTT.S156143

    Figure Lengend Snippet: SIRT4 suppresses EMT through promoting E-cadherin expression. ( A ) SIRT4 was overexpressed or knocked down in MKN-45 or HGC-27 cells. After transfection for 48 h, the mRNA level of E-cadherin was detected using qRT-PCR. SIRT4 vs vector and siSIRT4 vs SCR, * p

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) TRIzol reagent (Invitrogen) was used to extract total RNA from tissues or cells according to the manufacturer’s instructions.

    Techniques: Expressing, Transfection, Quantitative RT-PCR, Plasmid Preparation

    SIRT4 suppresses GC cell proliferation. ( A ) MKN-45 and HGC-27 were transfected with vector or SIRT4, scramble siRNA (SCR), or SIRT4 siRNA (siSIRT4). After transfection for 48 h, the mRNA level of SIRT4 was determined using qRT-PCR. SIRT4 vs vector and siSIRT4 vs SCR, * p

    Journal: OncoTargets and therapy

    Article Title: SIRT4 acts as a tumor suppressor in gastric cancer by inhibiting cell proliferation, migration, and invasion

    doi: 10.2147/OTT.S156143

    Figure Lengend Snippet: SIRT4 suppresses GC cell proliferation. ( A ) MKN-45 and HGC-27 were transfected with vector or SIRT4, scramble siRNA (SCR), or SIRT4 siRNA (siSIRT4). After transfection for 48 h, the mRNA level of SIRT4 was determined using qRT-PCR. SIRT4 vs vector and siSIRT4 vs SCR, * p

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) TRIzol reagent (Invitrogen) was used to extract total RNA from tissues or cells according to the manufacturer’s instructions.

    Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR

    KMP 1 specifically recognized CD 44 epitope located on the cell surface in bladder cancer. (A) To investigate the epitope recognized by KMP 1, the supernatants after EJ cell lysis were applied to protein A‐Sepharose column with KMP 1. Two peaks were obtained from the eluates by Sephacryl S200. (B) Antigen–antibody complex peak and antibody excess peak obtained from the eluates were resolved by 10% SDS – PAGE and stained with silver. (C) qRT – PCR analysis of CD 44 expression in EJ cells that were transfected with pS uper‐puro‐ CD 44‐sh RNA (sh CD 44‐ EJ ) or the empty pS uper‐puro vector (shCtrl‐ EJ ). (D) Western blot imagines indicated that the band using KMP 1 as a primary antibody was markedly thinned in the CD 44‐silenced EJ cells versus the shCtrl‐ EJ cells. (E) sh CD 44‐ EJ and shCtrl‐ EJ cells were analyzed by flow cytometry after staining with KMP 1, and nontransfected EJ cells were stained with mIgG as a negative control. (F) ELISA analysis of KMP 1 recognized CD 44 levels in shCtrl or sh CD 44‐ EJ cells. Each bar is expressed as the mean ± standard deviation of three experiments. * P

    Journal: Cancer Medicine

    Article Title: A novel monoclonal antibody KMP1 has potential antitumor activity of bladder cancer by blocking CD44 in vivo and in vitro

    doi: 10.1002/cam4.1446

    Figure Lengend Snippet: KMP 1 specifically recognized CD 44 epitope located on the cell surface in bladder cancer. (A) To investigate the epitope recognized by KMP 1, the supernatants after EJ cell lysis were applied to protein A‐Sepharose column with KMP 1. Two peaks were obtained from the eluates by Sephacryl S200. (B) Antigen–antibody complex peak and antibody excess peak obtained from the eluates were resolved by 10% SDS – PAGE and stained with silver. (C) qRT – PCR analysis of CD 44 expression in EJ cells that were transfected with pS uper‐puro‐ CD 44‐sh RNA (sh CD 44‐ EJ ) or the empty pS uper‐puro vector (shCtrl‐ EJ ). (D) Western blot imagines indicated that the band using KMP 1 as a primary antibody was markedly thinned in the CD 44‐silenced EJ cells versus the shCtrl‐ EJ cells. (E) sh CD 44‐ EJ and shCtrl‐ EJ cells were analyzed by flow cytometry after staining with KMP 1, and nontransfected EJ cells were stained with mIgG as a negative control. (F) ELISA analysis of KMP 1 recognized CD 44 levels in shCtrl or sh CD 44‐ EJ cells. Each bar is expressed as the mean ± standard deviation of three experiments. * P

    Article Snippet: The efficiency of CD44 silencing after transfection was examined on a 7900HT quantitative real‐time polymerase chain reaction (qRT–PCR) System (Applied Biosystems, Foster City, CA, USA). β ‐Actin (Sigma) was used as the internal control.

    Techniques: Lysis, SDS Page, Staining, Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Western Blot, Flow Cytometry, Cytometry, Negative Control, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Expression of proliferation and apoptosis associated genes and proteins. a mRNA expression of Akt, Erk, Casp3 and Casp9 using qRT-PCR; b protein expression of p-Akt, p-Erk, pho-Casp3 and pho-Casp9; c gray value of p-Akt, p-Erk, pho-Casp3 and pho-Casp9 using Western blotting. * P

    Journal: Cancer Cell International

    Article Title: Effect of long non-coding RNA Gas5 on proliferation, migration, invasion and apoptosis of colorectal cancer HT-29 cell line

    doi: 10.1186/s12935-017-0478-7

    Figure Lengend Snippet: Expression of proliferation and apoptosis associated genes and proteins. a mRNA expression of Akt, Erk, Casp3 and Casp9 using qRT-PCR; b protein expression of p-Akt, p-Erk, pho-Casp3 and pho-Casp9; c gray value of p-Akt, p-Erk, pho-Casp3 and pho-Casp9 using Western blotting. * P

    Article Snippet: Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) Trizol reagent (Life Technologies, Grand Island, NY, USA) was used to extract RNA from CRC and adjacent normal tissues (50–100 mg) by high glucose DMEM with fetal bovine serum (FBS; 10%).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    The relative expression of Gas5 in HT-29 cells among the blank, NC and pcDNA-Gas5 groups. a relative mRNA expression of Gas5 using qRT-PCR; b relative protein expression of Gas5; c gray value of Gas5 using Western blotting (1 represents the blank group; 2 the NC group; and 3 the pcDNA-Gas5 group). * P

    Journal: Cancer Cell International

    Article Title: Effect of long non-coding RNA Gas5 on proliferation, migration, invasion and apoptosis of colorectal cancer HT-29 cell line

    doi: 10.1186/s12935-017-0478-7

    Figure Lengend Snippet: The relative expression of Gas5 in HT-29 cells among the blank, NC and pcDNA-Gas5 groups. a relative mRNA expression of Gas5 using qRT-PCR; b relative protein expression of Gas5; c gray value of Gas5 using Western blotting (1 represents the blank group; 2 the NC group; and 3 the pcDNA-Gas5 group). * P

    Article Snippet: Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) Trizol reagent (Life Technologies, Grand Island, NY, USA) was used to extract RNA from CRC and adjacent normal tissues (50–100 mg) by high glucose DMEM with fetal bovine serum (FBS; 10%).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: HFD feeding increased the translocation to the nucleus of p65 and expression of NFκB‐related genes in Irf3 −/− mice. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expressions of phospho‐IKKα∕β, IKKβ, phospho‐p65, p65, and IκBα were assessed by western blot and normalized to GAPDH, n = 4 per genotype. (B) Formalin‐fixed paraffin‐embedded sections of liver were deparaffinized, and localization of phospho‐p65 was assessed by immunohistochemistry. Images were acquired at 40×. White arrows indicate nonparenchymal cells, and black arrows indicate hepatocytes. Images are representative of triplicate images from eight mice per treatment group. Values represent means ± SEM. (C) Nuclear fractions were isolated from livers, and equal amounts of protein were assessed by western blot and probed against phosphor‐Ser 536 p65‐NFκB. Lamin A/C was used as a nuclei marker, and GAPDH was used as a cytosolic marker. (D) Expressions of CCL5, CXCL10, CXCL2, and CCL3 were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values with different superscripts are significantly different from each other, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Translocation Assay, Expressing, Mouse Assay, Western Blot, Formalin-fixed Paraffin-Embedded, Immunohistochemistry, Isolation, Marker, Quantitative RT-PCR

    Irf3 S1/S1 mice reduced HFD‐induced liver injury, steatosis, and markers of inflammation. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) ALT and AST activities were measured in plasma, n = 12 per genotype. (C) Hepatic triglyceride content was measured in whole‐liver homogenates, n = 12 per genotype. (D) Paraffin‐embedded liver sections were stained with hematoxylin and eosin. Images were acquired using 10× and 40× objectives, n = 4 per genotype. Expressions of (E) TNFα, (F) IL‐1β, and (G) MCP‐1 mRNA were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Irf3 S1/S1 mice reduced HFD‐induced liver injury, steatosis, and markers of inflammation. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) ALT and AST activities were measured in plasma, n = 12 per genotype. (C) Hepatic triglyceride content was measured in whole‐liver homogenates, n = 12 per genotype. (D) Paraffin‐embedded liver sections were stained with hematoxylin and eosin. Images were acquired using 10× and 40× objectives, n = 4 per genotype. Expressions of (E) TNFα, (F) IL‐1β, and (G) MCP‐1 mRNA were analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Mouse Assay, AST Assay, Staining, Quantitative RT-PCR

    HFD enhanced CD45 + infiltration in Irf3 −/− but not in wild‐type and Irf3 S1/S1 livers. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of CD45 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. (B) Hepatic CD45 + leukocytes were gated in isolated liver NPCs by flow cytometry, and total number of cells were quantified in isolated liver NPCs, n = 4 per genotype. (C) Paraffin‐embedded liver sections were stained for CD45. Images were acquired using a 20× objective, n = 4 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: HFD enhanced CD45 + infiltration in Irf3 −/− but not in wild‐type and Irf3 S1/S1 livers. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of CD45 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. (B) Hepatic CD45 + leukocytes were gated in isolated liver NPCs by flow cytometry, and total number of cells were quantified in isolated liver NPCs, n = 4 per genotype. (C) Paraffin‐embedded liver sections were stained for CD45. Images were acquired using a 20× objective, n = 4 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Isolation, Flow Cytometry, Cytometry, Staining

    Enhanced markers of hepatocellular death and fibrosis in Irf3 −/− mice compared with wild‐type and Irf3 S1/S1 mice after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) Paraffin‐embedded liver sections were stained for M30, a caspase‐dependent cleavage product of cytokeratin 18. Images were acquired using 10× and 40× objectives, and the positive area was quantified, n = 4 per genotype. (C,D) Paraffin‐embedded liver sections were stained with sirius red. Images were acquired using a 40× objective, and the positive area was quantified, n = 4 per genotype. (E) Expression of Col1α1 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Enhanced markers of hepatocellular death and fibrosis in Irf3 −/− mice compared with wild‐type and Irf3 S1/S1 mice after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A,B) Paraffin‐embedded liver sections were stained for M30, a caspase‐dependent cleavage product of cytokeratin 18. Images were acquired using 10× and 40× objectives, and the positive area was quantified, n = 4 per genotype. (C,D) Paraffin‐embedded liver sections were stained with sirius red. Images were acquired using a 40× objective, and the positive area was quantified, n = 4 per genotype. (E) Expression of Col1α1 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 8 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

    Techniques: Mouse Assay, Staining, Expressing, Quantitative RT-PCR

    Hepatic IRF3 activation after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of IFIT1 and IFIT3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. (B) Expression of IRF3 was assessed by western blot and normalized to GAPDH, n = 4 per genotype. (C) Expression of IRF3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Journal: Hepatology Communications

    Article Title: Nontranscriptional Activity of Interferon Regulatory Factor 3 Protects Mice From High‐Fat Diet‐Induced Liver Injury

    doi: 10.1002/hep4.1441

    Figure Lengend Snippet: Hepatic IRF3 activation after HFD feeding. Male C57BL/6, Irf3 −/− , and Irf3 S1/S1 mice were fed an HFD or chow for 12 weeks. (A) Expression of IFIT1 and IFIT3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. (B) Expression of IRF3 was assessed by western blot and normalized to GAPDH, n = 4 per genotype. (C) Expression of IRF3 mRNA was analyzed by qRT‐PCR. Values were normalized to 18S and are shown as fold increase over the chow‐fed C57BL/6 control group, n = 12 per genotype. Values represent means ± SEM. Values with different alphabetical superscripts are significantly different, P

    Article Snippet: Two micrograms of liver RNA was reverse transcribed and analyzed with PowerSYBR quantitative real‐time polymerase chain reaction (qRT‐PCR) kits (Applied Biosystems, Foster City, CA) on a QuantStudio5 RT‐PCR Machine (Applied Biosystems).

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

    Osteogenic differentiation. The numbers in each plot indicate the different hUCMSC seeding densities on CPC-microbead-fiber scaffold, where 1k=1000. Real-time reverse transcription polymerase chain reaction on gene expression for (A) alkaline phosphatase

    Journal: Tissue Engineering. Part A

    Article Title: Effect of Cell Seeding Density on Proliferation and Osteodifferentiation of Umbilical Cord Stem Cells on Calcium Phosphate Cement-Fiber Scaffold

    doi: 10.1089/ten.tea.2011.0048

    Figure Lengend Snippet: Osteogenic differentiation. The numbers in each plot indicate the different hUCMSC seeding densities on CPC-microbead-fiber scaffold, where 1k=1000. Real-time reverse transcription polymerase chain reaction on gene expression for (A) alkaline phosphatase

    Article Snippet: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR, 7900HT; Applied Biosystems) was used. hUCMSCs attaching to CPC-microbead-fiber scaffold were cultured for 1, 4, 8, and 14 days in osteogenic media.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

    GATA3 inhibits epithelial–mesenchymal transition (EMT) in gastric cancer cells. Notes: ( A ) CTC-141 cells were transfected with vector, GATA3 and GATA3 (R298Q), and the relative expressions of EMT-associated genes were detected by qRT-PCR. * P

    Journal: Cancer Management and Research

    Article Title: Low expression of GATA3 promotes cell proliferation and metastasis in gastric cancer

    doi: 10.2147/CMAR.S147973

    Figure Lengend Snippet: GATA3 inhibits epithelial–mesenchymal transition (EMT) in gastric cancer cells. Notes: ( A ) CTC-141 cells were transfected with vector, GATA3 and GATA3 (R298Q), and the relative expressions of EMT-associated genes were detected by qRT-PCR. * P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) RNeasy Mini kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract total RNA from tissue samples and cells according to our protocol.

    Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR

    GATA3 is downregulated in gastric cancer tissues. Notes: ( A ) Total mRNA was extracted from 83 pairs of tumor tissue samples and adjacent normal tissue samples. qRT-PCR was performed to determine the relative mRNA level of GATA3. * P

    Journal: Cancer Management and Research

    Article Title: Low expression of GATA3 promotes cell proliferation and metastasis in gastric cancer

    doi: 10.2147/CMAR.S147973

    Figure Lengend Snippet: GATA3 is downregulated in gastric cancer tissues. Notes: ( A ) Total mRNA was extracted from 83 pairs of tumor tissue samples and adjacent normal tissue samples. qRT-PCR was performed to determine the relative mRNA level of GATA3. * P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) RNeasy Mini kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract total RNA from tissue samples and cells according to our protocol.

    Techniques: Quantitative RT-PCR

    GATA3 transcriptionally regulates ZEB1 in G cells. Notes: ( A ) CTC-141 cells were transfected with vector, GATA3 and GATA3 (R298Q), and the relative expressions of EMT-associated transcription factors were detected by qRT-PCR. ** P

    Journal: Cancer Management and Research

    Article Title: Low expression of GATA3 promotes cell proliferation and metastasis in gastric cancer

    doi: 10.2147/CMAR.S147973

    Figure Lengend Snippet: GATA3 transcriptionally regulates ZEB1 in G cells. Notes: ( A ) CTC-141 cells were transfected with vector, GATA3 and GATA3 (R298Q), and the relative expressions of EMT-associated transcription factors were detected by qRT-PCR. ** P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) RNeasy Mini kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract total RNA from tissue samples and cells according to our protocol.

    Techniques: Transfection, Plasmid Preparation, Quantitative RT-PCR

    Overexpression of ZEB1 suppresses the migratory and invasive behavior of gastric cancer cells. Notes: ( A ) ZEB1 was knocked down in CTC-141 cells. After transfection for 48 h, the expression of ZEB1 was determined by qRT-PCR. * P

    Journal: Cancer Management and Research

    Article Title: Low expression of GATA3 promotes cell proliferation and metastasis in gastric cancer

    doi: 10.2147/CMAR.S147973

    Figure Lengend Snippet: Overexpression of ZEB1 suppresses the migratory and invasive behavior of gastric cancer cells. Notes: ( A ) ZEB1 was knocked down in CTC-141 cells. After transfection for 48 h, the expression of ZEB1 was determined by qRT-PCR. * P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) RNeasy Mini kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract total RNA from tissue samples and cells according to our protocol.

    Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR

    GATA3 is downregulated in GC cells. Notes: ( A ) Expression of GATA3 in GC cell lines, CTC-141 and MKN45, and human normal gastric mucosal cell line GES-1 was detected by qRT-PCR and Western blotting, respectively. * P

    Journal: Cancer Management and Research

    Article Title: Low expression of GATA3 promotes cell proliferation and metastasis in gastric cancer

    doi: 10.2147/CMAR.S147973

    Figure Lengend Snippet: GATA3 is downregulated in GC cells. Notes: ( A ) Expression of GATA3 in GC cell lines, CTC-141 and MKN45, and human normal gastric mucosal cell line GES-1 was detected by qRT-PCR and Western blotting, respectively. * P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) RNeasy Mini kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract total RNA from tissue samples and cells according to our protocol.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot