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  • 94
    Bio-Rad qrt pcr analysis
    EOMES expression is increased in D*V embryonic comb tissue. Results of <t>qRT-PCR</t> analysis demonstrating increased EOMES expression in D*V embryonic comb tissue whereas CMC1 and AZI2 , located nearby the duplicated region, do not show any significant change in expression. Bar graphs mean ±sem, ANOVA, *P
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    Bio-Rad quantitative real time pcr qrt pcr analysis
    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) <t>qRT-PCR</t> analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
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    Bio-Rad cfx manager software
    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) <t>qRT-PCR</t> analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
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    Bio-Rad cfx96 real time pcr system
    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) <t>qRT-PCR</t> analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
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    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) <t>qRT-PCR</t> analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p
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    Bio-Rad quantitative real time polymerase chain reaction qrt pcr analysis
    Validation of the DGE analysis by quantitative real-time polymerase chain reaction <t>(qRT-PCR).</t> Error bars represent the standard deviations of qRT-PCR signals (n = 3).
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Analysis, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 90/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad cfx 96 real time machine
    Validation of the DGE analysis by quantitative real-time polymerase chain reaction <t>(qRT-PCR).</t> Error bars represent the standard deviations of qRT-PCR signals (n = 3).
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    Bio-Rad cfx connect real time pcr system
    Validation of the DGE analysis by quantitative real-time polymerase chain reaction <t>(qRT-PCR).</t> Error bars represent the standard deviations of qRT-PCR signals (n = 3).
    Cfx Connect Real Time Pcr System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 763 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    EOMES expression is increased in D*V embryonic comb tissue. Results of qRT-PCR analysis demonstrating increased EOMES expression in D*V embryonic comb tissue whereas CMC1 and AZI2 , located nearby the duplicated region, do not show any significant change in expression. Bar graphs mean ±sem, ANOVA, *P

    Journal: PLoS Genetics

    Article Title: A Genomic Duplication is Associated with Ectopic Eomesodermin Expression in the Embryonic Chicken Comb and Two Duplex-comb Phenotypes

    doi: 10.1371/journal.pgen.1004947

    Figure Lengend Snippet: EOMES expression is increased in D*V embryonic comb tissue. Results of qRT-PCR analysis demonstrating increased EOMES expression in D*V embryonic comb tissue whereas CMC1 and AZI2 , located nearby the duplicated region, do not show any significant change in expression. Bar graphs mean ±sem, ANOVA, *P

    Article Snippet: The qRT-PCR analysis was performed using CFX96 SyBr Green Supermix (Bio-Rad) with primers designed by using Primer Express v2.0 (ABI), checked for PCR efficiency, linear dynamic range and specificity.

    Techniques: Expressing, Quantitative RT-PCR

    TOR kinase phosphorylates and activates E2Fa a , Ectopic E2Fa activation of S-phase genes requires glucose-TOR signalling in leaf cells. WT or tor protoplasts expressing E2Fa-HA or S6K1-FLAG were treated without or with rapamycin (Rap) or antimycin A (AMA). QRT-PCR analyses. P-T449 indicates endogenous TOR kinase activity. Protein blot analysis (inset). b-c , TOR kinase directly phosphorylates E2Fa and 4E-BP1. Torin1 specifically inhibits TOR kinase. Staurosporine (Stau) inhibits S6K1 kinase. d , TOR directly interacts with E2Fa by immunoprecipitation (IP) and Western (W) blot analysis.

    Journal: Nature

    Article Title: Glc-TOR signalling leads transcriptome reprogramming and meristem activation

    doi: 10.1038/nature12030

    Figure Lengend Snippet: TOR kinase phosphorylates and activates E2Fa a , Ectopic E2Fa activation of S-phase genes requires glucose-TOR signalling in leaf cells. WT or tor protoplasts expressing E2Fa-HA or S6K1-FLAG were treated without or with rapamycin (Rap) or antimycin A (AMA). QRT-PCR analyses. P-T449 indicates endogenous TOR kinase activity. Protein blot analysis (inset). b-c , TOR kinase directly phosphorylates E2Fa and 4E-BP1. Torin1 specifically inhibits TOR kinase. Staurosporine (Stau) inhibits S6K1 kinase. d , TOR directly interacts with E2Fa by immunoprecipitation (IP) and Western (W) blot analysis.

    Article Snippet: All qRT-PCR analyses were performed by CFX96 real time PCR detection system with iQ SYBR green supermix (Bio-Rad).

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

    Auxin and cytokinin signalling and root stem-cell maintenance are decoupled from TOR activation a-b , Auxin signalling. c-d , Cytokinin signalling. Rapamycin (Rap), mitochondrial blocker (AMA). Primary auxin and cytokinin marker genes were activated by 1 h of indole-3-acetic acid (IAA) or trans-zeatin (tZ) treatment, and analysed by qRT-PCR. Means ± s.d., n=3. DR5::GFP or TCS::GFP was activated by 6 h of IAA or tZ treatment. Scale bar, 50 µm. e, f , Root stem-cell maintenance is TOR independent. PLT1::GFP , root stem-cell marker; WOX5::GFP : root quiescent centre marker. Scale bar, 20 µm.

    Journal: Nature

    Article Title: Glc-TOR signalling leads transcriptome reprogramming and meristem activation

    doi: 10.1038/nature12030

    Figure Lengend Snippet: Auxin and cytokinin signalling and root stem-cell maintenance are decoupled from TOR activation a-b , Auxin signalling. c-d , Cytokinin signalling. Rapamycin (Rap), mitochondrial blocker (AMA). Primary auxin and cytokinin marker genes were activated by 1 h of indole-3-acetic acid (IAA) or trans-zeatin (tZ) treatment, and analysed by qRT-PCR. Means ± s.d., n=3. DR5::GFP or TCS::GFP was activated by 6 h of IAA or tZ treatment. Scale bar, 50 µm. e, f , Root stem-cell maintenance is TOR independent. PLT1::GFP , root stem-cell marker; WOX5::GFP : root quiescent centre marker. Scale bar, 20 µm.

    Article Snippet: All qRT-PCR analyses were performed by CFX96 real time PCR detection system with iQ SYBR green supermix (Bio-Rad).

    Techniques: Activation Assay, Marker, Quantitative RT-PCR

    Glucose-TOR signalling orchestrates transcriptome reprogramming a-b , Glucose-TOR activated genes. c-d , Glucose-TOR repressed genes. 3DAG WT or tor seedlings were treated without or with glucose for 2 h. Hierarchical clustering analysis of glucose-TOR genes and five independent datasets (Glc2h, Glc4h, Sucrose, LowCO 2 -light, lowCO 2 -dark). Deep-pink/blue bar indicates novel glucose-TOR genes. The enriched functional categories highlighted in bold ( Supplementary Tables 1, 3, 4 ). e , Hierarchical clustering analysis of glucose-TOR genes (Glc) and cell cycle genes (G1, S, G2, M) 33 . f , Glucose-TOR activated genes overlap with E2Fa target genes. g , Glucose-TOR activates S-phase genes. QRT-PCR analyses. Means ± s.d., n=3. *P

    Journal: Nature

    Article Title: Glc-TOR signalling leads transcriptome reprogramming and meristem activation

    doi: 10.1038/nature12030

    Figure Lengend Snippet: Glucose-TOR signalling orchestrates transcriptome reprogramming a-b , Glucose-TOR activated genes. c-d , Glucose-TOR repressed genes. 3DAG WT or tor seedlings were treated without or with glucose for 2 h. Hierarchical clustering analysis of glucose-TOR genes and five independent datasets (Glc2h, Glc4h, Sucrose, LowCO 2 -light, lowCO 2 -dark). Deep-pink/blue bar indicates novel glucose-TOR genes. The enriched functional categories highlighted in bold ( Supplementary Tables 1, 3, 4 ). e , Hierarchical clustering analysis of glucose-TOR genes (Glc) and cell cycle genes (G1, S, G2, M) 33 . f , Glucose-TOR activated genes overlap with E2Fa target genes. g , Glucose-TOR activates S-phase genes. QRT-PCR analyses. Means ± s.d., n=3. *P

    Article Snippet: All qRT-PCR analyses were performed by CFX96 real time PCR detection system with iQ SYBR green supermix (Bio-Rad).

    Techniques: Functional Assay, Gas Chromatography, Quantitative RT-PCR

    TOR kinase controls the activity of E2Fa in transcriptional activation a , TOR kinase phosphorylates the N-terminal domain of E2Fa. b , TOR kinase phosphorylation is critical for E2Fa activation of S-phase genes. c , E2Fa-DNA binding is not affected by TOR kinase phosphorylation. ChIP-qPCR analyses with P (promoter) or G (gene body) primers. Stars, putative E2Fa-binding motifs. Error bars (n=2). d-e , Glucose responses is diminished in e2fa root meristems. Scale bar, 1 mm or 20 µm. QRT-PCR analyses. f , Model of leaf-root coordination in glucose-TOR signalling. GSH, GLUTATHIONE, RGF , ROOT GROWTH FACTOR, UPB1 , UPBEAT1. Means ± s.d., n=3. *P

    Journal: Nature

    Article Title: Glc-TOR signalling leads transcriptome reprogramming and meristem activation

    doi: 10.1038/nature12030

    Figure Lengend Snippet: TOR kinase controls the activity of E2Fa in transcriptional activation a , TOR kinase phosphorylates the N-terminal domain of E2Fa. b , TOR kinase phosphorylation is critical for E2Fa activation of S-phase genes. c , E2Fa-DNA binding is not affected by TOR kinase phosphorylation. ChIP-qPCR analyses with P (promoter) or G (gene body) primers. Stars, putative E2Fa-binding motifs. Error bars (n=2). d-e , Glucose responses is diminished in e2fa root meristems. Scale bar, 1 mm or 20 µm. QRT-PCR analyses. f , Model of leaf-root coordination in glucose-TOR signalling. GSH, GLUTATHIONE, RGF , ROOT GROWTH FACTOR, UPB1 , UPBEAT1. Means ± s.d., n=3. *P

    Article Snippet: All qRT-PCR analyses were performed by CFX96 real time PCR detection system with iQ SYBR green supermix (Bio-Rad).

    Techniques: Activity Assay, Activation Assay, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Expression analysis of five genes in cotton anthers by qRT-PCR. The housekeeping gene 18S was used as an internal control; H276A, CMS line; H276B, maintainer line; and H276A/H268, F1 plants from the descendant of restorer H268 hybridized with H276A. Error bars represent standard deviation (n = 3). Double asterisk represents significance at 1% probability

    Journal: Biological Research

    Article Title: RNA editing analysis of ATP synthase genes in the cotton cytoplasmic male sterile line H276A

    doi: 10.1186/s40659-019-0212-0

    Figure Lengend Snippet: Expression analysis of five genes in cotton anthers by qRT-PCR. The housekeeping gene 18S was used as an internal control; H276A, CMS line; H276B, maintainer line; and H276A/H268, F1 plants from the descendant of restorer H268 hybridized with H276A. Error bars represent standard deviation (n = 3). Double asterisk represents significance at 1% probability

    Article Snippet: qRT-PCR analysis of ATP synthase genes Relative quantification of five genes in the three cotton lines were conducted by real-time qRT-PCR analysis with a C1000 TouchTM Thermal Cycler (Bio-Rad, USA) with TransStartR Tip Green qPCR SuperMix (Trans, China).

    Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

    Mediator-independent and TAF6 and AF9-dependent interaction of TFIID and SEC in mammalian cells. A. Stable knockdown of TAF6 in 293T cells using TAF6-specific shRNA. Knockdown efficiency was tested by western blotting (upper panel) as well as RNA analysis by qRT-PCR. B. Stable knockdown of AF9 in 293T cells using AF9-specific shRNA. Knockdown efficiency was tested by western blotting (upper panel) as well as RNA analysis by qRT-PCR. C. Schematics of experimental strategy that has been employed for experiments mentioned in the panel D and E. D. Immunoblot analysis showing the effect of TAF6 knockdown on association of other TAF and SEC subunits with ectopically expressed FLAG-TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. E. Immunoblot analysis showing the effect of AF9 knockdown on association of other TAF and SEC subunits with ectopically expressed TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. F. Stable knockdown of Med26 in 293T cells using Med26-specific shRNA. Knockdown efficiency was tested by western blotting. G. Immunoblot analysis showing the effect of Med26 knockdown on association of other TAF and SEC subunits with ectopically expressed FLAG-TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. The left panel shows experimental strategy that was used for this assay. H. Immunoblot analysis showing the effect of Med26 knockdown on association of other TAF and SEC subunits with ectopically expressed EAF1. EAF1-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. The left panel shows experimental strategy that was used for this assay.

    Journal: Cell reports

    Article Title: Multivalent role of human TFIID in recruiting elongation components at the promoter proximal region for transcriptional control

    doi: 10.1016/j.celrep.2019.01.012

    Figure Lengend Snippet: Mediator-independent and TAF6 and AF9-dependent interaction of TFIID and SEC in mammalian cells. A. Stable knockdown of TAF6 in 293T cells using TAF6-specific shRNA. Knockdown efficiency was tested by western blotting (upper panel) as well as RNA analysis by qRT-PCR. B. Stable knockdown of AF9 in 293T cells using AF9-specific shRNA. Knockdown efficiency was tested by western blotting (upper panel) as well as RNA analysis by qRT-PCR. C. Schematics of experimental strategy that has been employed for experiments mentioned in the panel D and E. D. Immunoblot analysis showing the effect of TAF6 knockdown on association of other TAF and SEC subunits with ectopically expressed FLAG-TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. E. Immunoblot analysis showing the effect of AF9 knockdown on association of other TAF and SEC subunits with ectopically expressed TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. F. Stable knockdown of Med26 in 293T cells using Med26-specific shRNA. Knockdown efficiency was tested by western blotting. G. Immunoblot analysis showing the effect of Med26 knockdown on association of other TAF and SEC subunits with ectopically expressed FLAG-TBP. TBP-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. The left panel shows experimental strategy that was used for this assay. H. Immunoblot analysis showing the effect of Med26 knockdown on association of other TAF and SEC subunits with ectopically expressed EAF1. EAF1-associated proteins were pulled-down using anti-FLAG M2 beads and proteins were identified by immunoblotting using specific antibodies. The left panel shows experimental strategy that was used for this assay.

    Article Snippet: 1ug of RNA was reverse transcribed using verso cDNA synthesis kit (Thermo Scientific) using manufacturer's protocol. cDNA was diluted 25X for quantitative real-time PCR (qRT-PCR) analyses. qRT-PCR was performed using Universal SYBR green supermix (BIORAD).

    Techniques: Size-exclusion Chromatography, shRNA, Western Blot, Quantitative RT-PCR

    TFIID-mediated recruitment of SEC at promoter proximal region is required for basal level as well as inducible expression of target genes. A. qRT-PCR analysis showing effect of TAF6 and AF9 knockdown on basal level expression of target genes when compared to control scramble knockdown cells. Statistical significance for RNA analysis in TAF6 and AF9 knockdown samples are shown over control (Scramble) sample. B-D: ChIP analysis showing recruitment of several factors at promoter proximal region of indicated target genes upon knockdown of TAF6 and AF9. Statistical significance for ChIP analysis in TAF6 and AF9 knockdown samples are shown over control (Scramble) sample. E. qRT-PCR analysis showing effect of AF9 knockdown on induced expression of target p21 gene at different time points after doxorubicin treatment. F. ChIP analysis showing effect of AF9 knockdown on recruitment of target factors at promoter proximal and ~4kb downstream (coding) region of p21 gene after 16 hrs of doxorubicin treatment. Statistical significance for ChIP analysis in AF9 knockdown sample are shown over control (Scramble) sample. PP denotes promoter proximal region. All of our RNA expression and ChIP data represents mean ± SEM., a minimum of two biological and three PCR replicates. Statistical analysis was performed using one-tailed Student's t test wherein * denotes p ≤0.05, ** denotes p ≤0.01, *** denotes p ≤0.001, and ns denotes not significant.

    Journal: Cell reports

    Article Title: Multivalent role of human TFIID in recruiting elongation components at the promoter proximal region for transcriptional control

    doi: 10.1016/j.celrep.2019.01.012

    Figure Lengend Snippet: TFIID-mediated recruitment of SEC at promoter proximal region is required for basal level as well as inducible expression of target genes. A. qRT-PCR analysis showing effect of TAF6 and AF9 knockdown on basal level expression of target genes when compared to control scramble knockdown cells. Statistical significance for RNA analysis in TAF6 and AF9 knockdown samples are shown over control (Scramble) sample. B-D: ChIP analysis showing recruitment of several factors at promoter proximal region of indicated target genes upon knockdown of TAF6 and AF9. Statistical significance for ChIP analysis in TAF6 and AF9 knockdown samples are shown over control (Scramble) sample. E. qRT-PCR analysis showing effect of AF9 knockdown on induced expression of target p21 gene at different time points after doxorubicin treatment. F. ChIP analysis showing effect of AF9 knockdown on recruitment of target factors at promoter proximal and ~4kb downstream (coding) region of p21 gene after 16 hrs of doxorubicin treatment. Statistical significance for ChIP analysis in AF9 knockdown sample are shown over control (Scramble) sample. PP denotes promoter proximal region. All of our RNA expression and ChIP data represents mean ± SEM., a minimum of two biological and three PCR replicates. Statistical analysis was performed using one-tailed Student's t test wherein * denotes p ≤0.05, ** denotes p ≤0.01, *** denotes p ≤0.001, and ns denotes not significant.

    Article Snippet: 1ug of RNA was reverse transcribed using verso cDNA synthesis kit (Thermo Scientific) using manufacturer's protocol. cDNA was diluted 25X for quantitative real-time PCR (qRT-PCR) analyses. qRT-PCR was performed using Universal SYBR green supermix (BIORAD).

    Techniques: Size-exclusion Chromatography, Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, RNA Expression, Polymerase Chain Reaction, One-tailed Test

    Genes cyclically regulated by JH through Met—the effects of AAs, 20E, EcR and Met on genes cyclically repressed by Met. (A) Venn diagram showing genes that are up regulated in late post eclosion (LPE) and early post blood meal (EGs) periods and that are down regulated in Met knocked-down (imet down) fat body tissues. (B) Venn diagram showing genes that are up regulated in early post eclosion (EPE) and late-mid post blood meal (LMGs) periods and that are up regulated in Met knocked-down (imet up) fat body tissues. (C) Venn diagram showing genes that are up regulated in early post eclosion (EPE) and late-mid post blood meal (LMGs) periods and that are up regulated in Met knocked-down (imet up) fat body tissues. (D) Relative expression of the gene, AAEL002781, Galactokinase detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet); injecting double stranded RNA for the Luciferase gene (iluc) served as the control. (E) Relative expression of the same gene detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E). (F) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR).

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Genes cyclically regulated by JH through Met—the effects of AAs, 20E, EcR and Met on genes cyclically repressed by Met. (A) Venn diagram showing genes that are up regulated in late post eclosion (LPE) and early post blood meal (EGs) periods and that are down regulated in Met knocked-down (imet down) fat body tissues. (B) Venn diagram showing genes that are up regulated in early post eclosion (EPE) and late-mid post blood meal (LMGs) periods and that are up regulated in Met knocked-down (imet up) fat body tissues. (C) Venn diagram showing genes that are up regulated in early post eclosion (EPE) and late-mid post blood meal (LMGs) periods and that are up regulated in Met knocked-down (imet up) fat body tissues. (D) Relative expression of the gene, AAEL002781, Galactokinase detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet); injecting double stranded RNA for the Luciferase gene (iluc) served as the control. (E) Relative expression of the same gene detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E). (F) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR).

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, Luciferase, In Vitro

    Effects of AAs, 20E, insulin and HR3 on YPP genes. (A) Relative expression of gene—AAEL006563, Carboxypeptidase, detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression in tissues subjected IVFBC in culture media without (NT) and with amino acids (AA), with amino acids plus 20E (AA+20E) and after the withdrawal of 20E (20E WD). (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). (E) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (NT) and with amino acids (AA), with amino acids and Insulin (AA+INS), Insulin and 20E (INS+20E), amino acids plus 20E (AA+20E), and amino acids plus 20E and Insulin (AA+20E+INS). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Effects of AAs, 20E, insulin and HR3 on YPP genes. (A) Relative expression of gene—AAEL006563, Carboxypeptidase, detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression in tissues subjected IVFBC in culture media without (NT) and with amino acids (AA), with amino acids plus 20E (AA+20E) and after the withdrawal of 20E (20E WD). (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). (E) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (NT) and with amino acids (AA), with amino acids and Insulin (AA+INS), Insulin and 20E (INS+20E), amino acids plus 20E (AA+20E), and amino acids plus 20E and Insulin (AA+20E+INS). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    Effects of AAs, 20E and JH on representative LGs. (A) Relative expression of gene—AALE004328 –Origin recognition complex, detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (C) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiment. All expressions calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Effects of AAs, 20E and JH on representative LGs. (A) Relative expression of gene—AALE004328 –Origin recognition complex, detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (C) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiment. All expressions calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    Effects of AAs, 20E and HR3 on EMGs. (A) Relative expression of gene—AAEL001433, fgf receptor activating protein detected by qRT-PCR, in tissues subjected to in- vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression in tissues subjected to IVFBC in culture media without (NT) and with amino acids (AA), with amino acids plus 20E (AA+20E) and after the withdrawal of 20E (20E WD). (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Effects of AAs, 20E and HR3 on EMGs. (A) Relative expression of gene—AAEL001433, fgf receptor activating protein detected by qRT-PCR, in tissues subjected to in- vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression in tissues subjected to IVFBC in culture media without (NT) and with amino acids (AA), with amino acids plus 20E (AA+20E) and after the withdrawal of 20E (20E WD). (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    Genes cyclically activated by JH through Met—Functional group enrichment and the effects of AAs, 20E and JH. (A) Venn diagram showing genes that are up regulated in late post eclosion (LPE) and late post blood meal (LGs) periods and that are down regulated in Met knocked-down (imet down) fat body tissues. (B-D) Comparison of functional categories viz. (B) Information storage and Processing, (C) Metabolism and (D) Cellular Processes and Signalling, that constitute the late genes (LGs) and cyclical genes (CGs) using the inNOG database. (E-F) Relative expression of gene—AAEL001171, tRNA-dihydrouridine synthase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media, (E) without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E); (F) without (JH-) and with (JH+) juvenile hormone. (G) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet); injecting double stranded RNA for the Luciferase gene (iluc) served as the control. (H) Expression profile of the gene after the first blood meal. (I) Expression profile of the gene after the completion of the first reproductive cycle (egg laying) and post second blood meal. All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Genes cyclically activated by JH through Met—Functional group enrichment and the effects of AAs, 20E and JH. (A) Venn diagram showing genes that are up regulated in late post eclosion (LPE) and late post blood meal (LGs) periods and that are down regulated in Met knocked-down (imet down) fat body tissues. (B-D) Comparison of functional categories viz. (B) Information storage and Processing, (C) Metabolism and (D) Cellular Processes and Signalling, that constitute the late genes (LGs) and cyclical genes (CGs) using the inNOG database. (E-F) Relative expression of gene—AAEL001171, tRNA-dihydrouridine synthase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media, (E) without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E); (F) without (JH-) and with (JH+) juvenile hormone. (G) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet); injecting double stranded RNA for the Luciferase gene (iluc) served as the control. (H) Expression profile of the gene after the first blood meal. (I) Expression profile of the gene after the completion of the first reproductive cycle (egg laying) and post second blood meal. All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Functional Assay, Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    Effects of AAs, 20E, JH and HR3 on representative LMGs. (A) Relative expression of genes—AAEL003568, Threonine dehydratase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). (D) Relative expression in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (E) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B , C and E ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Effects of AAs, 20E, JH and HR3 on representative LMGs. (A) Relative expression of genes—AAEL003568, Threonine dehydratase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post HR3 knock-down (iHR3). (D) Relative expression in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (E) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes post Met knock-down (iMet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B , C and E ). All expression calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    Effects of AAs, 20E and JH on representative EGs. (A) Relative expression of AAEL002269, Purine nucleoside phosphorylase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes after knock-down of the JH receptor Met (imet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expressions calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Journal: PLoS Genetics

    Article Title: Regulation of Gene Expression Patterns in Mosquito Reproduction

    doi: 10.1371/journal.pgen.1005450

    Figure Lengend Snippet: Effects of AAs, 20E and JH on representative EGs. (A) Relative expression of AAEL002269, Purine nucleoside phosphorylase detected by qRT-PCR, in tissues subjected to in-vitro fat body culture (IVFBC) in culture media without (NT) and with amino acids (AA) and with amino acid plus 20E (AA+20E) (B) Relative expression of the same gene detected by qRT-PCR, in fat body tissues collected from female mosquitoes post EcR knock-down (iEcR). (C) Relative expression of the same gene in tissues subjected to IVFBC in culture media without (JH-) and with (JH+) juvenile hormone. (D) Relative expression detected by qRT-PCR, in fat body tissues collected from female mosquitoes after knock-down of the JH receptor Met (imet). Injecting double stranded RNA for the Luciferase gene (iluc) served as the control in the RNAi experiments ( B and D ). All expressions calculated against housekeeping gene RPS7. Data representative of three biological replicates, with three technical replicates and are illustrated as average ± SD, * P

    Article Snippet: qRT-PCR analysis qRT-PCR was performed using the iCycler iQ system (Bio-Rad, Hercules, CA and an IQ SYBR Green Supermix (Bio-Rad).

    Techniques: Atomic Absorption Spectroscopy, Expressing, Quantitative RT-PCR, In Vitro, Luciferase

    BACE-1 mRNA expression analysis in mouse brain microvessels and cultured endothelial cells. In freshly isolated mouse brain microvessels (MBMVs) and primary cultured mouse brain microvascular endothelial cells (MBMECs) mRNA expression analysis by qRT-PCR showed significant expression of BACE-1 using two different primer pairs. Whole brain mRNA served as a positive control for BACE-1 expression as neurons are known to express high levels of BACE-1. BACE-1 expression was present even in pure cultured endothelial cells that have no contamination from neurons thus indicating a specific expression of BACE-1. Cldn-5 served as a marker for endothelium, which was at much higher levels in cultured and freshly isolated brain microvascular endothelial cells. The Ct range for BACE-1 qRT-PCR was in the range 23–26 cycles with non-template control about 35 cycles indicating specificity of expression. Statistical significance was by One-way ANOVA followed by TUKEY-HSD test for multiple groups (n = 4 experiments, * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: BACE-1 is expressed in the blood–brain barrier endothelium and is upregulated in a murine model of Alzheimer’s disease

    doi: 10.1177/0271678X15606463

    Figure Lengend Snippet: BACE-1 mRNA expression analysis in mouse brain microvessels and cultured endothelial cells. In freshly isolated mouse brain microvessels (MBMVs) and primary cultured mouse brain microvascular endothelial cells (MBMECs) mRNA expression analysis by qRT-PCR showed significant expression of BACE-1 using two different primer pairs. Whole brain mRNA served as a positive control for BACE-1 expression as neurons are known to express high levels of BACE-1. BACE-1 expression was present even in pure cultured endothelial cells that have no contamination from neurons thus indicating a specific expression of BACE-1. Cldn-5 served as a marker for endothelium, which was at much higher levels in cultured and freshly isolated brain microvascular endothelial cells. The Ct range for BACE-1 qRT-PCR was in the range 23–26 cycles with non-template control about 35 cycles indicating specificity of expression. Statistical significance was by One-way ANOVA followed by TUKEY-HSD test for multiple groups (n = 4 experiments, * p

    Article Snippet: Analysis of the qRT-PCR data was performed with iQ5 2.1 software (Bio-Rad).

    Techniques: Expressing, Cell Culture, Isolation, Quantitative RT-PCR, Positive Control, Marker

    Expression analysis in brain microvessels from an AD mouse model and schematic for APP/Aβ processing at the BBB. (a) MBMVs from transgenic mice (hAPP SL ) over-expressing the 751 amino acid form of human amyloid precursor protein (hAPP) with London (V717I) and Swedish (KM670/671NL) mutations under the control of the murine Thy-1 promoter were compared with wild-type mice. We observed a four-fold increase of BACE-1 expression in the BBB microvessels isolated from hAPP SL mice when compared to age-matched wild-type mice suggesting an increase in the APP cleavage activity of BACE-1 at the BBB in the mutant animals. Endothelial marker genes such as VE-cadherin and tight junction molecules ZO-1, claudin-5 were unchanged whereas GLUT-1, the primary glucose transporter at the BBB was downregulated. Interestingly, the luminal Aβ transporter RAGE was upregulated in the AD mice suggesting that circulating Aβ could potentially contribute to the brain amyloidosis. Furthermore the luminally expressed p-glycoprotein (PgP) known to be involved in efflux of Aβ into the circulation was downregulated suggesting a decreased clearance of amyloid peptides from the brain. The abluminally located LRP-1 was upregulated, which is known to endocytose APP, also supporting an increase in the BBB BACE-1 activity in this AD model. Abluminal Aβ antibody transporter FcRN was unchanged. Statistical significance was obtained from three qRT-PCR experiments using six transgenic mice (10 months age) or age-matched wild-type animals (*** p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: BACE-1 is expressed in the blood–brain barrier endothelium and is upregulated in a murine model of Alzheimer’s disease

    doi: 10.1177/0271678X15606463

    Figure Lengend Snippet: Expression analysis in brain microvessels from an AD mouse model and schematic for APP/Aβ processing at the BBB. (a) MBMVs from transgenic mice (hAPP SL ) over-expressing the 751 amino acid form of human amyloid precursor protein (hAPP) with London (V717I) and Swedish (KM670/671NL) mutations under the control of the murine Thy-1 promoter were compared with wild-type mice. We observed a four-fold increase of BACE-1 expression in the BBB microvessels isolated from hAPP SL mice when compared to age-matched wild-type mice suggesting an increase in the APP cleavage activity of BACE-1 at the BBB in the mutant animals. Endothelial marker genes such as VE-cadherin and tight junction molecules ZO-1, claudin-5 were unchanged whereas GLUT-1, the primary glucose transporter at the BBB was downregulated. Interestingly, the luminal Aβ transporter RAGE was upregulated in the AD mice suggesting that circulating Aβ could potentially contribute to the brain amyloidosis. Furthermore the luminally expressed p-glycoprotein (PgP) known to be involved in efflux of Aβ into the circulation was downregulated suggesting a decreased clearance of amyloid peptides from the brain. The abluminally located LRP-1 was upregulated, which is known to endocytose APP, also supporting an increase in the BBB BACE-1 activity in this AD model. Abluminal Aβ antibody transporter FcRN was unchanged. Statistical significance was obtained from three qRT-PCR experiments using six transgenic mice (10 months age) or age-matched wild-type animals (*** p

    Article Snippet: Analysis of the qRT-PCR data was performed with iQ5 2.1 software (Bio-Rad).

    Techniques: Expressing, Transgenic Assay, Mouse Assay, Isolation, Activity Assay, Mutagenesis, Marker, Quantitative RT-PCR

    DNA demethylation of ROR2 , p14 , and p16 in DME-expressing cells is accompanied by gene reactivation. (A) Schematic diagram of analyzed genes. Each vertical bar represents a CpG dinucleotide. Position of ATG codon is indicated as a red rectangle. Green arrows show the location of pyrosequencing primers and yellow arrows the location of qMSP primers. (B) Methylation levels analyzed by bisulfite pyrosequencing; CpG sites are shown as bars filled with black to represent percentage methylation. (C) Methylation levels analyzed by qMSP (D) Gene expression levels analyzed by qRT-PCR. Analyses were performed in non-transfected DLD-1 cells and independent transfectants expressing WT DME (DME 2, DME 10, and DME 13), a catalytically inactive mutant version (mut 7 and mut 13) or cells transfected with the empty vector. Values are shown relative to those detected in non-transfected cells. Data are the mean ± SE of three independent experiments.

    Journal: Epigenetics

    Article Title: DNA methylation reprogramming of human cancer cells by expression of a plant 5-methylcytosine DNA glycosylase

    doi: 10.1080/15592294.2017.1414128

    Figure Lengend Snippet: DNA demethylation of ROR2 , p14 , and p16 in DME-expressing cells is accompanied by gene reactivation. (A) Schematic diagram of analyzed genes. Each vertical bar represents a CpG dinucleotide. Position of ATG codon is indicated as a red rectangle. Green arrows show the location of pyrosequencing primers and yellow arrows the location of qMSP primers. (B) Methylation levels analyzed by bisulfite pyrosequencing; CpG sites are shown as bars filled with black to represent percentage methylation. (C) Methylation levels analyzed by qMSP (D) Gene expression levels analyzed by qRT-PCR. Analyses were performed in non-transfected DLD-1 cells and independent transfectants expressing WT DME (DME 2, DME 10, and DME 13), a catalytically inactive mutant version (mut 7 and mut 13) or cells transfected with the empty vector. Values are shown relative to those detected in non-transfected cells. Data are the mean ± SE of three independent experiments.

    Article Snippet: Quantitative real time RT-PCR (qRT-PCR) analysis was performed in a CFX Connect™ Real-Time PCR Detection System (Biorad) by mixing cDNA (from 1 μg total RNA) with iQ™ SYBR Green Supermix (Biorad) and specific primers (Table S4).

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

    DME expression on three loci displays different types of methylation changes in DLD-1 cells. (A) Methylation levels analyzed by bisulfite pyrosequencing; Each vertical bar represents a CpG dinucleotide, and position of ATG codon is indicated as a red rectangle; green arrows show the location of pyrosequencing primers. Analyzed CpG sites are shown as bars filled with black to represent percentage methylation. (B) Gene expression levels analyzed by qRT-PCR. Values are shown relative to those detected in non-transfected cells. Data are the mean ± SE of three independent experiments.

    Journal: Epigenetics

    Article Title: DNA methylation reprogramming of human cancer cells by expression of a plant 5-methylcytosine DNA glycosylase

    doi: 10.1080/15592294.2017.1414128

    Figure Lengend Snippet: DME expression on three loci displays different types of methylation changes in DLD-1 cells. (A) Methylation levels analyzed by bisulfite pyrosequencing; Each vertical bar represents a CpG dinucleotide, and position of ATG codon is indicated as a red rectangle; green arrows show the location of pyrosequencing primers. Analyzed CpG sites are shown as bars filled with black to represent percentage methylation. (B) Gene expression levels analyzed by qRT-PCR. Values are shown relative to those detected in non-transfected cells. Data are the mean ± SE of three independent experiments.

    Article Snippet: Quantitative real time RT-PCR (qRT-PCR) analysis was performed in a CFX Connect™ Real-Time PCR Detection System (Biorad) by mixing cDNA (from 1 μg total RNA) with iQ™ SYBR Green Supermix (Biorad) and specific primers (Table S4).

    Techniques: Expressing, Methylation, Quantitative RT-PCR, Transfection

    qRT-PCR validation of the differentially expressed miRNAs and target genes from high-throughput sequencing analyses in tuberous root development of radish. a qRT-PCR validation of 11 miRNAs from high-throughput small RNA sequencing analyses. b qRT-PCR validation of six target genes from degradome analyses

    Journal: 3 Biotech

    Article Title: Identification of miRNAs and their targets in regulating tuberous root development in radish using small RNA and degradome analyses

    doi: 10.1007/s13205-018-1330-z

    Figure Lengend Snippet: qRT-PCR validation of the differentially expressed miRNAs and target genes from high-throughput sequencing analyses in tuberous root development of radish. a qRT-PCR validation of 11 miRNAs from high-throughput small RNA sequencing analyses. b qRT-PCR validation of six target genes from degradome analyses

    Article Snippet: Real-time quantitative PCR (qRT-PCR) analysis was used to validate the expression patterns of miRNAs and target genes using an IQ5 Real-Time PCR System (BIO-RAD, Hercules, CA, USA).

    Techniques: Quantitative RT-PCR, Next-Generation Sequencing, High Throughput Screening Assay, RNA Sequencing Assay

    MnSR-BI expression tissue distribution as determined by real-time qRT-PCR. Values are shown as mean ± SD ( n = 3). Bars with different letters represent significant differences ( P

    Journal: International Journal of Genomics

    Article Title: Scavenger Receptor Class B, Type I, a CD36 Related Protein in Macrobrachium nipponense: Characterization, RNA Interference, and Expression Analysis with Different Dietary Lipid Sources

    doi: 10.1155/2016/6325927

    Figure Lengend Snippet: MnSR-BI expression tissue distribution as determined by real-time qRT-PCR. Values are shown as mean ± SD ( n = 3). Bars with different letters represent significant differences ( P

    Article Snippet: The SYBR Premix Ex Taq™ Kit (Takara) was used for real-time quantitative RT-PCR (qRT-PCR) analysis in a CFX96™ Real-Time System (Bio-Rad, USA).

    Techniques: Expressing, Quantitative RT-PCR

    Real-time quantitative PCR analysis of the MnSR-BI transcript expression in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA. Samples were obtained 48 h (a) and 96 h (b) after dsRNA injection and analyzed by real-time qRT-PCR. Bars indicate mean ± SD ( n = 3). ∗ P

    Journal: International Journal of Genomics

    Article Title: Scavenger Receptor Class B, Type I, a CD36 Related Protein in Macrobrachium nipponense: Characterization, RNA Interference, and Expression Analysis with Different Dietary Lipid Sources

    doi: 10.1155/2016/6325927

    Figure Lengend Snippet: Real-time quantitative PCR analysis of the MnSR-BI transcript expression in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA. Samples were obtained 48 h (a) and 96 h (b) after dsRNA injection and analyzed by real-time qRT-PCR. Bars indicate mean ± SD ( n = 3). ∗ P

    Article Snippet: The SYBR Premix Ex Taq™ Kit (Takara) was used for real-time quantitative RT-PCR (qRT-PCR) analysis in a CFX96™ Real-Time System (Bio-Rad, USA).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Injection, Quantitative RT-PCR

    A two-gene stemness signature, SOX2 and ALDH2, discriminates muscle-invasive from non-muscle-invasive BC A. qRT-PCR analysis of mRNA expression of pluripotency-related transcription factors (NANOG, POU5F1, SOX2), drug resistance-related genes (ABCG2, ABCB1, ALDH1A1, ALDH2, ALDH7A1) and urothelial basal cell-related markers (CD44, CD47 and KRT14) in BC samples divided in three groups based on tumor grade: non-muscle-invasive, low grade; non-muscle-invasive, high grade and muscle-invasive. The expression levels of target genes in each sample were normalized to three housekeeping genes and are shown in scatter plots * p

    Journal: Oncotarget

    Article Title: Functional and molecular characterization of cancer stem-like cells in bladder cancer: a potential signature for muscle-invasive tumors

    doi:

    Figure Lengend Snippet: A two-gene stemness signature, SOX2 and ALDH2, discriminates muscle-invasive from non-muscle-invasive BC A. qRT-PCR analysis of mRNA expression of pluripotency-related transcription factors (NANOG, POU5F1, SOX2), drug resistance-related genes (ABCG2, ABCB1, ALDH1A1, ALDH2, ALDH7A1) and urothelial basal cell-related markers (CD44, CD47 and KRT14) in BC samples divided in three groups based on tumor grade: non-muscle-invasive, low grade; non-muscle-invasive, high grade and muscle-invasive. The expression levels of target genes in each sample were normalized to three housekeeping genes and are shown in scatter plots * p

    Article Snippet: Quantitative real-time PCR analysis (qRT-PCR) was performed using a Quantitect SYBR Green qRT-PCR kit (Biorad) for NANOG, POU5F1 (OCT4), SOX2, ABCG2 (BCRP), ABCB1 (PGP), ALDH1A1, ALDH7A1 and ALDH2 in a Bio-Rad CFX 96 Thermal Cycler (Bio-Rad Laboratories, CA, United States). qRT-PCR amplification of CD47, CD44 and KRT14 genes was performed using the SYBR-Green I Master (Roche) according to the manufacturer's instructions in a LightCycler 480 II system (Roche Diagnostics, Mannheim, Germany). mRNA expression was normalized to three housekeeping genes: 18S, GAPDH and HRPT-1 using the δδCt method and Bio-Rad CFX Manager™ 3.0 software.

    Techniques: Quantitative RT-PCR, Expressing

    BC cells contain a population of sphere-forming cells with stem-like properties A. Parental monolayer HT-1376 and UM-UC3 cells form spherical colonies generated from a single cell, when cultured in serum-free DMEM/F12 medium, supplemented with bFGF, EGF and B27 supplement in Matrigel coated plates after 11 days. Scale bars = 50 μm. B. qRT-PCR analysis expression of pluripotency-related transcription factors (NANOG, POU5F1, SOX2), drug-resistance related genes (ABCG2, ABCB1, ALDH1A1, ALDH2, ALDH7A1) and urothelial basal cell-related markers (CD44, CD47 and KRT14) by qRT–PCR. The graph shows the fold change in gene expression in spheres relative to parental cell line that was set as 1 (mean + SEM, n = 4). C. Representative immunofluorescence staining of CD44 and CD47 monoclonal antibodies in spheres and corresponding parental cells. Cell nuclei were counterstained with Hoechst 33258 (in blue). The graph shows the mean fluorescence intensities in the confocal micrographs ( n = 3). * p

    Journal: Oncotarget

    Article Title: Functional and molecular characterization of cancer stem-like cells in bladder cancer: a potential signature for muscle-invasive tumors

    doi:

    Figure Lengend Snippet: BC cells contain a population of sphere-forming cells with stem-like properties A. Parental monolayer HT-1376 and UM-UC3 cells form spherical colonies generated from a single cell, when cultured in serum-free DMEM/F12 medium, supplemented with bFGF, EGF and B27 supplement in Matrigel coated plates after 11 days. Scale bars = 50 μm. B. qRT-PCR analysis expression of pluripotency-related transcription factors (NANOG, POU5F1, SOX2), drug-resistance related genes (ABCG2, ABCB1, ALDH1A1, ALDH2, ALDH7A1) and urothelial basal cell-related markers (CD44, CD47 and KRT14) by qRT–PCR. The graph shows the fold change in gene expression in spheres relative to parental cell line that was set as 1 (mean + SEM, n = 4). C. Representative immunofluorescence staining of CD44 and CD47 monoclonal antibodies in spheres and corresponding parental cells. Cell nuclei were counterstained with Hoechst 33258 (in blue). The graph shows the mean fluorescence intensities in the confocal micrographs ( n = 3). * p

    Article Snippet: Quantitative real-time PCR analysis (qRT-PCR) was performed using a Quantitect SYBR Green qRT-PCR kit (Biorad) for NANOG, POU5F1 (OCT4), SOX2, ABCG2 (BCRP), ABCB1 (PGP), ALDH1A1, ALDH7A1 and ALDH2 in a Bio-Rad CFX 96 Thermal Cycler (Bio-Rad Laboratories, CA, United States). qRT-PCR amplification of CD47, CD44 and KRT14 genes was performed using the SYBR-Green I Master (Roche) according to the manufacturer's instructions in a LightCycler 480 II system (Roche Diagnostics, Mannheim, Germany). mRNA expression was normalized to three housekeeping genes: 18S, GAPDH and HRPT-1 using the δδCt method and Bio-Rad CFX Manager™ 3.0 software.

    Techniques: Generated, Cell Culture, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Fluorescence

    qRT-PCR analysis of CTA expression in cohort 1 Data are expressed as means calculated out of assays run in triplicate. β-actin was used as internal reference to compute the δCt values.

    Journal: Oncotarget

    Article Title: Novel antigens in non-small cell lung cancer: SP17, AKAP4, and PTTG1 are potential immunotherapeutic targets

    doi:

    Figure Lengend Snippet: qRT-PCR analysis of CTA expression in cohort 1 Data are expressed as means calculated out of assays run in triplicate. β-actin was used as internal reference to compute the δCt values.

    Article Snippet: qRT-PCR qRT-PCR analysis was performed as previously described[ ], using an iCycler iQ Real Time PCR machine SYBR ® Green I Supermix (both from Bio-Rad Laboratories, Inc).

    Techniques: Quantitative RT-PCR, Expressing

    Proline affects the expression of CYCB1;1 in the root meristematic zone. a-p CYCB1::GUS expression, from 1 to 7 dag, in roots from CYCB1::GUS ( a, e, i, m ), p5cs1 p5cs2/P5CS2, CYCB1::GUS ( b, f, j, n ), proline-treated CYCB1::GUS ( c, g, k, q ) and proline-treated p5cs1 p5cs2/P5CS2, CYCB1::GUS ( d, h, l, p ). Bottom and top arrowheads show meristem size indicating the QC and, respectively, the TZ. Bars = 50 μm ( a-d ), 20 μm ( e-p ). q qRT-PCR of CYCB1;1 , at 3 and 5 dag, in root meristems of wild type (dark grey bar), p5cs1 p5cs2/P5CS2 (light grey bar), and proline-treated p5cs1 p5cs2/P5CS2 (grey bar), showing, at 3 dag, a strong downregulation of CYB1;1 expression in p5cs1 p5cs2/P5CS2 roots. The meristem-specific gene RCH1 was used as reference control to normalize the qRT-PCR

    Journal: BMC Plant Biology

    Article Title: Proline affects the size of the root meristematic zone in Arabidopsis

    doi: 10.1186/s12870-015-0637-8

    Figure Lengend Snippet: Proline affects the expression of CYCB1;1 in the root meristematic zone. a-p CYCB1::GUS expression, from 1 to 7 dag, in roots from CYCB1::GUS ( a, e, i, m ), p5cs1 p5cs2/P5CS2, CYCB1::GUS ( b, f, j, n ), proline-treated CYCB1::GUS ( c, g, k, q ) and proline-treated p5cs1 p5cs2/P5CS2, CYCB1::GUS ( d, h, l, p ). Bottom and top arrowheads show meristem size indicating the QC and, respectively, the TZ. Bars = 50 μm ( a-d ), 20 μm ( e-p ). q qRT-PCR of CYCB1;1 , at 3 and 5 dag, in root meristems of wild type (dark grey bar), p5cs1 p5cs2/P5CS2 (light grey bar), and proline-treated p5cs1 p5cs2/P5CS2 (grey bar), showing, at 3 dag, a strong downregulation of CYB1;1 expression in p5cs1 p5cs2/P5CS2 roots. The meristem-specific gene RCH1 was used as reference control to normalize the qRT-PCR

    Article Snippet: Real-time qRT-PCR analyses were carried out with a Bio-Rad iCycler iQ (Bio-Rad).

    Techniques: Expressing, Quantitative RT-PCR

    INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: INPP4B is upregulated by NPM1-mA in leukemia cells via ERK/Ets-1 signaling. ( a ) qRT-PCR analysis of INPP4B mRNA expression, ( b ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. ( c ) qRT-PCR analysis of INPP4B mRNA expression, ( d ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the THP-1 and K562 cells transduced with the plasmids expressing NPM1-wt or NPM1-mA. ( e ) qRT-PCR analysis of INPP4B mRNA expression, ( f ) western blotting analysis of INPP4B, p-SGK3 T320 , SGK3, p-Ets-1 and Ets-1 from the OCI-AML3 cells transfected with the control siRNA or siEts-1. ( g ) Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 and SGK3, ( h ) qRT-PCR analysis of INPP4B mRNA expression from the OCI-AML3 cells treated with different concentration of PD98059 (0, 10, 20 and 40 μM). i Western blotting analysis of p-ERK, ERK, p-Ets-1, Ets-1, INPP4B, p-SGK3 T320 , SGK3 and NPM1-mA from the NPM1-mA-silenced OCI-AML3 cells. Proteins were quantified using image software and normalized against β-actin. Data were represented as mean ± s.d. of three individual experiments. * p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed on an MJ Mini™ Gradient Thermal Cycler Real-Time PCR machine (Bio-Rad, CA, USA) with the SYBR Green reaction kit (KAPA Biosystems, MA, USA).

    Techniques: Quantitative RT-PCR, Expressing, Western Blot, Transduction, Transfection, Concentration Assay, Software

    INPP4B promotes cell proliferation in OCI-AML3 cells. ( a ) qRT-PCR and ( b ) western blotting analysis of INPP4B expression from the OCI-AML3 cells transduced with the control siRNA or siINPP4B. c CCK-8 assay analysis of cell proliferation activity in the siINPP4B transduced cells. d The OCI-AML3 cells stably infected with shRNA lentivirus targeting INPP4B were subjected to colony forming assays. e The INPP4B-silenced OCI-AML3 cells were transfected with the pEAK-Flag/INPP4B plasmids, western blotting analysis of INPP4B protein levels and quantified using image software normalized against β-actin. f CCK-8 assay analysis of cell proliferation in INPP4B-silenced OCI-AML3 cells, followed by Flag-INPP4B introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: INPP4B promotes cell proliferation in OCI-AML3 cells. ( a ) qRT-PCR and ( b ) western blotting analysis of INPP4B expression from the OCI-AML3 cells transduced with the control siRNA or siINPP4B. c CCK-8 assay analysis of cell proliferation activity in the siINPP4B transduced cells. d The OCI-AML3 cells stably infected with shRNA lentivirus targeting INPP4B were subjected to colony forming assays. e The INPP4B-silenced OCI-AML3 cells were transfected with the pEAK-Flag/INPP4B plasmids, western blotting analysis of INPP4B protein levels and quantified using image software normalized against β-actin. f CCK-8 assay analysis of cell proliferation in INPP4B-silenced OCI-AML3 cells, followed by Flag-INPP4B introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed on an MJ Mini™ Gradient Thermal Cycler Real-Time PCR machine (Bio-Rad, CA, USA) with the SYBR Green reaction kit (KAPA Biosystems, MA, USA).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transduction, CCK-8 Assay, Activity Assay, Stable Transfection, Infection, shRNA, Transfection, Software

    SGK3 is required for INPP4B-induced cell proliferation in OCI-AML3 cells. qRT-PCR ( a ) and western blotting ( b ) analysis of SGK3 expression from the OCI-AML3 cells transduced with the control siRNA or siSGK3. c CCK-8 assay analysis of cell proliferation activity in the siSGK3 transduced cells. d The INPP4B-silenced OCI-AML3 cells were transfected with the pCMV-Flag/SGK3 plasmids, western blotting analysis of SGK3, p-SGK3 T320 and INPP4B. Proteins were quantified using image software and normalized against β-actin. e CCK-8 assay analysis of cell proliferation in INPP4B-silenced OCI-AML3 cells, followed by Flag-SGK3 introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    Article Title: INPP4B promotes cell survival via SGK3 activation in NPM1-mutated leukemia

    doi: 10.1186/s13046-018-0675-9

    Figure Lengend Snippet: SGK3 is required for INPP4B-induced cell proliferation in OCI-AML3 cells. qRT-PCR ( a ) and western blotting ( b ) analysis of SGK3 expression from the OCI-AML3 cells transduced with the control siRNA or siSGK3. c CCK-8 assay analysis of cell proliferation activity in the siSGK3 transduced cells. d The INPP4B-silenced OCI-AML3 cells were transfected with the pCMV-Flag/SGK3 plasmids, western blotting analysis of SGK3, p-SGK3 T320 and INPP4B. Proteins were quantified using image software and normalized against β-actin. e CCK-8 assay analysis of cell proliferation in INPP4B-silenced OCI-AML3 cells, followed by Flag-SGK3 introduction. Data were represented as mean ± s.d. of three individual experiments. * p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) analysis was performed on an MJ Mini™ Gradient Thermal Cycler Real-Time PCR machine (Bio-Rad, CA, USA) with the SYBR Green reaction kit (KAPA Biosystems, MA, USA).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transduction, CCK-8 Assay, Activity Assay, Transfection, Software

    QRT-PCR analysis of genes responsible for B cell differentiation. Purified MZ B cells and FO B cells from control WT mice or CD40LBTg mice were cultured in vitro with a combination of IL-4, anti-IgM [F(ab′) 2 ], and anti-CD40 Ab or with CpGODN for

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

    Article Title: Constitutive CD40L Expression on B Cells Prematurely Terminates Germinal Center Response and Leads to Augmented Plasma Cell Production in T Cell Areas

    doi: 10.4049/jimmunol.0901689

    Figure Lengend Snippet: QRT-PCR analysis of genes responsible for B cell differentiation. Purified MZ B cells and FO B cells from control WT mice or CD40LBTg mice were cultured in vitro with a combination of IL-4, anti-IgM [F(ab′) 2 ], and anti-CD40 Ab or with CpGODN for

    Article Snippet: Quantitative RT-PCR (QRT-PCR) analysis was performed using primers from SABioscience with an iQ5 cycler (Bio-Rad, Hercules, CA).

    Techniques: Quantitative RT-PCR, Cell Differentiation, Purification, Mouse Assay, Cell Culture, In Vitro

    Validation of the DGE analysis by quantitative real-time polymerase chain reaction (qRT-PCR). Error bars represent the standard deviations of qRT-PCR signals (n = 3).

    Journal: PLoS ONE

    Article Title: RNA-seq approach to analysis of gene expression profiles in dark green islands and light green tissues of Cucumber mosaic virus-infected Nicotiana tabacum

    doi: 10.1371/journal.pone.0175391

    Figure Lengend Snippet: Validation of the DGE analysis by quantitative real-time polymerase chain reaction (qRT-PCR). Error bars represent the standard deviations of qRT-PCR signals (n = 3).

    Article Snippet: Quantitative real-time polymerase chain reaction validation Validation of the RNA-seq data for 12 different genes was performed using quantitative real-time polymerase chain reaction (qRT-PCR) analysis (Bio-Rad Laboratories, Hercules, CA, USA).

    Techniques: Real-time Polymerase Chain Reaction, Quantitative RT-PCR