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  • 99
    Thermo Fisher qrt pcr kit
    The caf5 gene is induced in a pap1 -dependent manner in response to LatA. <t>qRT-PCR</t> analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.
    Qrt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 959 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    qrt  (TaKaRa)
    99
    TaKaRa qrt
    The caf5 gene is induced in a pap1 -dependent manner in response to LatA. <t>qRT-PCR</t> analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.
    Qrt, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vazyme Biotech Co qrt supermix
    The caf5 gene is induced in a pap1 -dependent manner in response to LatA. <t>qRT-PCR</t> analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.
    Qrt Supermix, supplied by Vazyme Biotech Co, 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 qrt pcrs
    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using <t>qRT-PCRs.</t> (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.
    Qrt Pcrs, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1220 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vazyme Biotech Co hiscript qrt supermix
    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using <t>qRT-PCRs.</t> (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.
    Hiscript Qrt Supermix, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 89/100, based on 217 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    InvivoGen ifnr qrt primers
    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using <t>qRT-PCRs.</t> (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.
    Ifnr Qrt Primers, supplied by InvivoGen, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa qrt pcr
    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using <t>qRT-PCRs.</t> (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.
    Qrt Pcr, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 4531 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher quantitative real time pcr qrt pcr qrt pcr
    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with <t>qRT-PCR.</t> The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p
    Quantitative Real Time Pcr Qrt Pcr Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 277 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt pct primer
    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with <t>qRT-PCR.</t> The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p
    Qrt Pct Primer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Vazyme Biotech Co hisscript qrt supermix
    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with <t>qRT-PCR.</t> The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p
    Hisscript Qrt Supermix, supplied by Vazyme Biotech Co, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt pcr
    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with <t>qRT-PCR.</t> The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p
    Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 22478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    qrt  (Luminex)
    92
    Luminex qrt
    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with <t>qRT-PCR.</t> The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p
    Qrt, supplied by Luminex, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher 7900ht qrt pcr
    Sulf2 expression is altered in MCF-7 and MDA-MB-231 breast cancer cell lines. (A) The mRNA level of Sulf2 in MCF-7 shSulf2 was significantly lower than in MCF-7 NC. (B) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was significantly higher than in MDA-MB-231 vector. (C) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was 16-fold higher than in MCF-7 NC. (D) Sulf2 protein levels detected by western blot analyis were consistent with <t>qRT-PCR</t> results. * P
    7900ht Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene qrt pcr analysis
    Influence of DDX5 on T cell phenotypes in autoimmune disease models a , At 8 weeks after T cell transfer, LILP mononuclear cells were evaluated for amounts of IL-17A and IFNγ mRNA by <t>qRT-PCR.</t> Results are representative of two independent experiments. Each experiment was performed using large intestines from 3 animals in each condition. qRT-PCR was performed with two technical replicates. Graph shows mean ± s.d. * p
    Qrt Pcr Analysis, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa qrt pcr analysis
    ADAMTS5 and IGFBP5 are downregulated by miR-140 in CRC cells. a The mRNAs of ADAMTS5 and IGFBP5 contain putative binding sites of miR-140. b HCT116 and RKO cells were transfected with miR-140 mimic using oligofectamine. Oligofectamine alone (Control) and negative miRNA (NC) were the negative controls. The relative expression of miR-140 was determined by <t>qRT-PCR</t> after normalization to the internal control, RNU6B. * P
    Qrt Pcr Analysis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 2371 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher qrt pcr analysis
    PolyIC/dsRBEC induces the expression and secretion of pro-inflammatory cytokines in MDA-MB-468 cells. A) <t>qRT-PCR</t> analysis of IFN-β, CCL5, IP10 and TNFα mRNA expression following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 2 and 4 hours. Data were normalized to GAPDH and are expressed as fold change relative to vehicle-treated samples. A representative experiment out of 3 experiments is shown. Error bars represent RQ max. B) Protein levels of IFN-β, CCL5, IP10 and TNFα were measured by ELISA following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 24 hours. Values are averages of triplicate biological samples from one representative experiment. (***,P
    Qrt Pcr Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 11244 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad qrt pcr analyzer
    PolyIC/dsRBEC induces the expression and secretion of pro-inflammatory cytokines in MDA-MB-468 cells. A) <t>qRT-PCR</t> analysis of IFN-β, CCL5, IP10 and TNFα mRNA expression following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 2 and 4 hours. Data were normalized to GAPDH and are expressed as fold change relative to vehicle-treated samples. A representative experiment out of 3 experiments is shown. Error bars represent RQ max. B) Protein levels of IFN-β, CCL5, IP10 and TNFα were measured by ELISA following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 24 hours. Values are averages of triplicate biological samples from one representative experiment. (***,P
    Qrt Pcr Analyzer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    SABiosciences qrt pcr array
    Macrophages are recruited to the myocardium in lipotoxic cardiomyopathy. A : hearts from 4-wk-old ACS and NTG littermates were harvested and <t>qRT-PCR</t> was performed on total mRNA to assess the expression of macrophage markers. Graphs show mean expression
    Qrt Pcr Array, supplied by SABiosciences, used in various techniques. Bioz Stars score: 91/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Kapa Biosystems qrt pcr kit
    Spatial and temporal distribution of VvCCD1 , VvCCD4a and VvCCD4b transcripts in distinct grapevine tissues. <t>qRT-PCR</t> analysis of VvCCD1 , VvCCD4a and VvCCD4b in leaf, flower and three berry developmental stages. Data are expressed relative to pre-véraison berry stage and normalised to the housekeeping gene, VvEF1a . Relative changes in the total carotenoid and chlorophyll concentrations in the respective tissues are also shown.
    Qrt Pcr Kit, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt pcr primers
    Th2 cytokines suppress pro-caspase-1 without affecting levels of pro-IL-1β. (A) Taqman quantitative <t>RT-PCR</t> results showing mRNA expression of pro-IL-1β in THP-1s exposed to 100 μg/mL MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. B) Taqman <t>qRT-PCR</t> results showing mRNA levels of pro-caspase-1 in THP-1s exposed to MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. Statistical analysis was performed using a one-way ANOVA with a post hoc Tukey. ***P
    Qrt Pcr Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1082 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher qrt pcr probes
    Overexpression of CD86FLm provides a survival advantage against silencing of CD86 and CD28 but not against silencing of IRF4. (A) Representative histograms showing the levels of surface CD86 in 3 different cell lines when either CD28, CD86, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are silenced. Thin black histograms at left are unstained pLKO.1-infected controls. (B) Cell death measured at day 4 postinfection via annexin V staining, shown as percentage of pLKO.1-infected controls. (A-B) Data shown are from day 4 postinfection and representative of at least 3 independent experiments. (C) <t>qRT-PCR</t> was performed to determine levels of CD86 , CD28 , and IRF4 to compare the different cell lines. Data are normalized to β-actin as endogenous control and then compared relative to mRNA levels in pLKO.1 empty vector control. Data shown are mean ± SEM of at least 3 independent experiments. (D) Representative western blots showing levels of IRF4 in cells overexpressing CD86FLm or CD86TLm when either CD86 or CD28 are silenced. Lysates are from day 4 postinfection. (E) Cell death as measured by annexin V staining at day 4 postinfection in 8226-pCDNA3.1 or 8226-CD86FLm cells where CD86 or IRF4 was silenced; shown as percentage of pLKO.1-infected controls. (F) Representative western blots showing levels of IRF4 and β-actin in lysates from experiments in panel E. Data shown are mean ± SEM of at least 3 independent experiments. * P
    Qrt Pcr Probes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The caf5 gene is induced in a pap1 -dependent manner in response to LatA. qRT-PCR analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.

    Journal: G3: Genes|Genomes|Genetics

    Article Title: Latrunculin A-Induced Perturbation of the Actin Cytoskeleton Mediates Pap1p-Dependent Induction of the Caf5p Efflux Pump in Schizosaccharomyces pombe

    doi: 10.1534/g3.116.037903

    Figure Lengend Snippet: The caf5 gene is induced in a pap1 -dependent manner in response to LatA. qRT-PCR analysis of caf5 and trr1 expression (relative to gpd1 ) in response to 0.3 μM LatA treatment in (A) wild-type, (B) pap1 Δ, and (C) act1-R183A , D184A backgrounds. The level of caf5 and trr1 transcripts were normalized to that of the internal control gpd1 . The normalized level of the transcripts in LatA-treated cells is shown relative to that of DMSO controls. The mean ± SE for three biological replicates is shown. DMSO, dimethyl sulfoxide; LatA, latrunculin A; qRT-PCR, quantitative real time-polymerase chain reaction.

    Article Snippet: qRT-PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) according to the supplier’s protocol. cDNA synthesis was performed with 1 µg of total RNA using the SuperScript III First Strand Synthesis Supermix for qRT-PCR kit (Invitrogen), according to the supplier’s protocol.

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

    Isolation of villi and crypt epithelial cells of the jejunum from BL-6 WT and PepT1 KO mice. ( a ) Total RNAs were extracted from the different fractions collected from BL-6 WT mice using the low-temperature method, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. ( b ) Total RNAs were extracted from different fractions collected from PepT1 KO mice, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. ( c ) Pictures of selected villi and crypts fractions were taken with a Nikon Eclipse TS100 microscope at 10× magnifications. Scale bar, 50 μm. ( d ) Total RNAs were extracted from selected villus and crypt fractions from BL-6 WT and PepT1 KO mice. The expression levels of Muc2, Lgr5 and mPepT1 in the villi (blue bars) and crypts (magenta) of BL-6 WT and PepT1 KO mice were further assessed by qRT-PCR (n = 5; and ** P

    Journal: Scientific Reports

    Article Title: PepT1 Expression Helps Maintain Intestinal Homeostasis by Mediating the Differential Expression of miRNAs along the Crypt-Villus Axis

    doi: 10.1038/srep27119

    Figure Lengend Snippet: Isolation of villi and crypt epithelial cells of the jejunum from BL-6 WT and PepT1 KO mice. ( a ) Total RNAs were extracted from the different fractions collected from BL-6 WT mice using the low-temperature method, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. ( b ) Total RNAs were extracted from different fractions collected from PepT1 KO mice, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. ( c ) Pictures of selected villi and crypts fractions were taken with a Nikon Eclipse TS100 microscope at 10× magnifications. Scale bar, 50 μm. ( d ) Total RNAs were extracted from selected villus and crypt fractions from BL-6 WT and PepT1 KO mice. The expression levels of Muc2, Lgr5 and mPepT1 in the villi (blue bars) and crypts (magenta) of BL-6 WT and PepT1 KO mice were further assessed by qRT-PCR (n = 5; and ** P

    Article Snippet: RNA extraction and real-time reverse transcription-PCR Total RNA was extracted from mouse small intestine with the RNeasy Mini kit (Qiagen, Valencia, CA), according to the manufacturer’s instructions, and yield and quality verified. cDNA was generated from total RNA using the maxima first-strand cDNA synthesis kit (Thermo Scientific, Glen-Burnie, MD). cDNA of miRNAs was generated from total RNA preparations using NCode miRNA first-Strand cDNA synthesis and qRT-PCR kits (Invitrogen, Carlsbad, CA).

    Techniques: Isolation, Mouse Assay, Expressing, Marker, Quantitative RT-PCR, Microscopy

    PepT1 knockout reduces mouse body weight and the size of intestinal microvilli. ( a ) Genotyping was performed on PepT1 KO and BL-6 WT mice. Homozygous PepT1 KO mice and appropriate BL-6 WT controls were used in our studies. ( b , c ) PepT1 expression levels in the small intestines of PepT1 KO and BL-6 WT mice were detected by Western blotting ( b ) and qRT-PCR ( c ). ( d ) Representative H E-stained sections of small intestines from BL-6 WT and PepT1 KO mice showed no significant gross morphology difference. Original magnification, 20×. Scale bars, 50 μm. ( e ) Body weights measured at the indicated ages were significantly higher in BL-6 WT mice than in PepT1 KO mice (PepT1 KO: n = 9/group, BL-6 WT: n = 9/group *** P

    Journal: Scientific Reports

    Article Title: PepT1 Expression Helps Maintain Intestinal Homeostasis by Mediating the Differential Expression of miRNAs along the Crypt-Villus Axis

    doi: 10.1038/srep27119

    Figure Lengend Snippet: PepT1 knockout reduces mouse body weight and the size of intestinal microvilli. ( a ) Genotyping was performed on PepT1 KO and BL-6 WT mice. Homozygous PepT1 KO mice and appropriate BL-6 WT controls were used in our studies. ( b , c ) PepT1 expression levels in the small intestines of PepT1 KO and BL-6 WT mice were detected by Western blotting ( b ) and qRT-PCR ( c ). ( d ) Representative H E-stained sections of small intestines from BL-6 WT and PepT1 KO mice showed no significant gross morphology difference. Original magnification, 20×. Scale bars, 50 μm. ( e ) Body weights measured at the indicated ages were significantly higher in BL-6 WT mice than in PepT1 KO mice (PepT1 KO: n = 9/group, BL-6 WT: n = 9/group *** P

    Article Snippet: RNA extraction and real-time reverse transcription-PCR Total RNA was extracted from mouse small intestine with the RNeasy Mini kit (Qiagen, Valencia, CA), according to the manufacturer’s instructions, and yield and quality verified. cDNA was generated from total RNA using the maxima first-strand cDNA synthesis kit (Thermo Scientific, Glen-Burnie, MD). cDNA of miRNAs was generated from total RNA preparations using NCode miRNA first-Strand cDNA synthesis and qRT-PCR kits (Invitrogen, Carlsbad, CA).

    Techniques: Knock-Out, Mouse Assay, Expressing, Western Blot, Quantitative RT-PCR, Staining

    The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using qRT-PCRs. (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.

    Journal: Scientific Reports

    Article Title: Genome-wide identification of Arabidopsis long noncoding RNAs in response to the blue light

    doi: 10.1038/s41598-020-63187-1

    Figure Lengend Snippet: The validation of lncRNA- and miRNA-seq data. (A – F) The expression patterns of indicated lncRNAs in Col seedlings under blue light treatments for 2 h or 8 h using qRT-PCRs. (G) Northern blots show the levels of miRNAs in Col seedlings under blue light treatments for 2 h or 8 h. U6 serves as a loading control.

    Article Snippet: The primers used for qRT-PCRs are listed in Table S2 supplemental information.

    Techniques: Expressing, Northern Blot

    Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with qRT-PCR. The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p

    Journal: Development & Reproduction

    Article Title: Expression Analysis of Interferon-Stimulated Gene 15 in the Rock Bream Oplegnathus fasciatus against Rock Bream Iridovirus (RSIV) Challenge

    doi: 10.12717/DR.2017.21.4.371

    Figure Lengend Snippet: Temporal distribution of ISG15 mRNA in various tissues following RSIV infection. (A) The expression level of ISG15 was determined in the (A) kidney, (B) muscle, and (C) spleen with qRT-PCR. The samples were analyzed at 0, 3, 6, 12, 24, and 72 hours post-injection. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) qRT-PCR (ABI 7500, Applied Biosystems) was performed on rock bream samples to quantify the expression of ISG15.

    Techniques: Infection, Expressing, Quantitative RT-PCR, Injection

    Temporal expression analysis of ISG15 following rock bream iridovirus (RSIV) infection in whole fish. (A) Reverse transcription (RT)-PCR was performed with 1 μg of total RNA using ISG15-qRT-1F and 1R primers. Beta-actin was included in all reactions to verify equal complementary DNA concentrations in the PCR reaction and on the gel. (B) qRT-PCR analysis of ISG15 in rock bream following RSIV infection at 0, 3, 6, 12, 24, and 72 h. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p

    Journal: Development & Reproduction

    Article Title: Expression Analysis of Interferon-Stimulated Gene 15 in the Rock Bream Oplegnathus fasciatus against Rock Bream Iridovirus (RSIV) Challenge

    doi: 10.12717/DR.2017.21.4.371

    Figure Lengend Snippet: Temporal expression analysis of ISG15 following rock bream iridovirus (RSIV) infection in whole fish. (A) Reverse transcription (RT)-PCR was performed with 1 μg of total RNA using ISG15-qRT-1F and 1R primers. Beta-actin was included in all reactions to verify equal complementary DNA concentrations in the PCR reaction and on the gel. (B) qRT-PCR analysis of ISG15 in rock bream following RSIV infection at 0, 3, 6, 12, 24, and 72 h. Each experiment was performed in triplicate and the expression levels of beta actin and ISG15 at 0 h were set as 1. Asterisks indicate statistically significant differences ( * p

    Article Snippet: Quantitative real-time PCR (qRT-PCR) qRT-PCR (ABI 7500, Applied Biosystems) was performed on rock bream samples to quantify the expression of ISG15.

    Techniques: Expressing, Infection, Fluorescence In Situ Hybridization, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Quantitative RT-PCR

    Sulf2 expression is altered in MCF-7 and MDA-MB-231 breast cancer cell lines. (A) The mRNA level of Sulf2 in MCF-7 shSulf2 was significantly lower than in MCF-7 NC. (B) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was significantly higher than in MDA-MB-231 vector. (C) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was 16-fold higher than in MCF-7 NC. (D) Sulf2 protein levels detected by western blot analyis were consistent with qRT-PCR results. * P

    Journal: Oncology Reports

    Article Title: Sulfatase 2 promotes breast cancer progression through regulating some tumor-related factors

    doi: 10.3892/or.2015.4525

    Figure Lengend Snippet: Sulf2 expression is altered in MCF-7 and MDA-MB-231 breast cancer cell lines. (A) The mRNA level of Sulf2 in MCF-7 shSulf2 was significantly lower than in MCF-7 NC. (B) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was significantly higher than in MDA-MB-231 vector. (C) The mRNA level of Sulf2 in MDA-MB-231 Sulf2 was 16-fold higher than in MCF-7 NC. (D) Sulf2 protein levels detected by western blot analyis were consistent with qRT-PCR results. * P

    Article Snippet: The mRNA level was determined by 7900HT qRT-PCR (Applied Biosystems, Foster City, CA, USA) using SYBR® Green Real-time PCR Master Mix (Takara, Shiga, Japan).

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

    The results of the PCR microarray screening confirmed using real-time PCR. (A) The results of the PCR microarray screening were verified using qRT-PCR in MCF-7 NC and MCF-7 shSulf2 cell lines. (B) The results of the PCR microarray screening were confirmed using qRT-PCR in MDA-MB-231 vector and MDA-MB-231 Sulf2 cells. (C) The Sulf2 protein expression of four genes were changed with alteration of Sulf2 expression. * P

    Journal: Oncology Reports

    Article Title: Sulfatase 2 promotes breast cancer progression through regulating some tumor-related factors

    doi: 10.3892/or.2015.4525

    Figure Lengend Snippet: The results of the PCR microarray screening confirmed using real-time PCR. (A) The results of the PCR microarray screening were verified using qRT-PCR in MCF-7 NC and MCF-7 shSulf2 cell lines. (B) The results of the PCR microarray screening were confirmed using qRT-PCR in MDA-MB-231 vector and MDA-MB-231 Sulf2 cells. (C) The Sulf2 protein expression of four genes were changed with alteration of Sulf2 expression. * P

    Article Snippet: The mRNA level was determined by 7900HT qRT-PCR (Applied Biosystems, Foster City, CA, USA) using SYBR® Green Real-time PCR Master Mix (Takara, Shiga, Japan).

    Techniques: Polymerase Chain Reaction, Microarray, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Multiple Displacement Amplification, Plasmid Preparation, Expressing

    Influence of DDX5 on T cell phenotypes in autoimmune disease models a , At 8 weeks after T cell transfer, LILP mononuclear cells were evaluated for amounts of IL-17A and IFNγ mRNA by qRT-PCR. Results are representative of two independent experiments. Each experiment was performed using large intestines from 3 animals in each condition. qRT-PCR was performed with two technical replicates. Graph shows mean ± s.d. * p

    Journal: Nature

    Article Title: DDX5 and its associated lncRNA Rmrp modulate Th17 cell effector functions

    doi: 10.1038/nature16193

    Figure Lengend Snippet: Influence of DDX5 on T cell phenotypes in autoimmune disease models a , At 8 weeks after T cell transfer, LILP mononuclear cells were evaluated for amounts of IL-17A and IFNγ mRNA by qRT-PCR. Results are representative of two independent experiments. Each experiment was performed using large intestines from 3 animals in each condition. qRT-PCR was performed with two technical replicates. Graph shows mean ± s.d. * p

    Article Snippet: Isolated RNA was used in qRT-PCR analysis (Stratagene) to quantify enrichment of RMRP and depletion of other cellular RNAs.

    Techniques: Quantitative RT-PCR

    Rmrp localization at RORγt-occupied genes and role in RORγt-DDX5 assembly a , RORγt association with immunoprecipitated (IP) DDX5 in polarized Th17 cells. IB, immunoblot. Representative of three independent experiments. b , Rmrp quantification by qRT-PCR in RORγt immunoprecipitates from polarized Th17 cells. Representative of two independent experiments with two technical replicates. c , Rmrp requirement for ATP-dependent in vitro interaction of recombinant GST-DDX5 and His-RORγt. Representative of three independent experiments. For gel source data (a,c), see Supplementary Figure 1 . d , Rmrp occupancy at RORγt genomic target loci in polarized Th17 cells. Rmrp ChIRP-qPCR amplicons (bottom) are indicated in IGV browser view of RORγt ChIP at the Il17 locus (top). Data from 2–4 experiments with two technical replicates. e , Model for DDX5-Rmrp complex recruitment to RORγt-occupied chromatin in Th17 cells. Graphs show mean ± s.d. ** p

    Journal: Nature

    Article Title: DDX5 and its associated lncRNA Rmrp modulate Th17 cell effector functions

    doi: 10.1038/nature16193

    Figure Lengend Snippet: Rmrp localization at RORγt-occupied genes and role in RORγt-DDX5 assembly a , RORγt association with immunoprecipitated (IP) DDX5 in polarized Th17 cells. IB, immunoblot. Representative of three independent experiments. b , Rmrp quantification by qRT-PCR in RORγt immunoprecipitates from polarized Th17 cells. Representative of two independent experiments with two technical replicates. c , Rmrp requirement for ATP-dependent in vitro interaction of recombinant GST-DDX5 and His-RORγt. Representative of three independent experiments. For gel source data (a,c), see Supplementary Figure 1 . d , Rmrp occupancy at RORγt genomic target loci in polarized Th17 cells. Rmrp ChIRP-qPCR amplicons (bottom) are indicated in IGV browser view of RORγt ChIP at the Il17 locus (top). Data from 2–4 experiments with two technical replicates. e , Model for DDX5-Rmrp complex recruitment to RORγt-occupied chromatin in Th17 cells. Graphs show mean ± s.d. ** p

    Article Snippet: Isolated RNA was used in qRT-PCR analysis (Stratagene) to quantify enrichment of RMRP and depletion of other cellular RNAs.

    Techniques: Immunoprecipitation, Quantitative RT-PCR, In Vitro, Recombinant, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation

    DDX5 co-regulates a subset of RORγt transcriptional targets in polarized Th17 cells a , Venn diagram of distinct and overlapping genes regulated by RORγt and/or DDX5, as determined from RNA-seq studies. b , Ingenuity Pathway Analysis of DDX5- and RORγt-coregulated genes. c , IGV browser view showing biological replicate RNA-seq coverage tracks of control, DDX5-Tko, or RORγt-deficient in vitro polarized Th17 cell samples at the Il17a, Il22, Ddx5 , and Rorc loci. d , Independent qRT-PCR validation of RNA-seq results confirming effects of DDX5 deletion on RORγt target gene expression. Graph shows mean ± s.d.

    Journal: Nature

    Article Title: DDX5 and its associated lncRNA Rmrp modulate Th17 cell effector functions

    doi: 10.1038/nature16193

    Figure Lengend Snippet: DDX5 co-regulates a subset of RORγt transcriptional targets in polarized Th17 cells a , Venn diagram of distinct and overlapping genes regulated by RORγt and/or DDX5, as determined from RNA-seq studies. b , Ingenuity Pathway Analysis of DDX5- and RORγt-coregulated genes. c , IGV browser view showing biological replicate RNA-seq coverage tracks of control, DDX5-Tko, or RORγt-deficient in vitro polarized Th17 cell samples at the Il17a, Il22, Ddx5 , and Rorc loci. d , Independent qRT-PCR validation of RNA-seq results confirming effects of DDX5 deletion on RORγt target gene expression. Graph shows mean ± s.d.

    Article Snippet: Isolated RNA was used in qRT-PCR analysis (Stratagene) to quantify enrichment of RMRP and depletion of other cellular RNAs.

    Techniques: RNA Sequencing Assay, In Vitro, Quantitative RT-PCR, Expressing

    ncRNAs enriched in DDX5 and RORγt RIP-seq studies a , DDX5-Tko cells were transduced with WT or helicase-mutant DDX5 and evaluated for DDX5 expression by immunofluorescence (left) and immunoblot (right) with anti-DDX5 antibody. For gel source data, see Supplementary Figure 1 . b , Venn diagram of ncRNAs detected by deep sequencing following co-immunoprecipitation (RIP-seq) of ribosome-depleted Th17 cell lysates with anti-DDX5 and anti-RORγt antibodies. c , Abundance of top ncRNAs enriched in DDX5 and RORγt immunoprecipitates from polarized Th17 cell lysates depleted of ribosomes. Top panel indicates abundance of the ncRNAs in total lysate. d , Conventional RIP-qRT-PCR experiments to compare Rmrp association with DDX5 in Th17 and developing thymocytes. Results are representative of three independent experiments. Each experiment was performed with two technical replicates. Graph shows mean ± s.d. ** p

    Journal: Nature

    Article Title: DDX5 and its associated lncRNA Rmrp modulate Th17 cell effector functions

    doi: 10.1038/nature16193

    Figure Lengend Snippet: ncRNAs enriched in DDX5 and RORγt RIP-seq studies a , DDX5-Tko cells were transduced with WT or helicase-mutant DDX5 and evaluated for DDX5 expression by immunofluorescence (left) and immunoblot (right) with anti-DDX5 antibody. For gel source data, see Supplementary Figure 1 . b , Venn diagram of ncRNAs detected by deep sequencing following co-immunoprecipitation (RIP-seq) of ribosome-depleted Th17 cell lysates with anti-DDX5 and anti-RORγt antibodies. c , Abundance of top ncRNAs enriched in DDX5 and RORγt immunoprecipitates from polarized Th17 cell lysates depleted of ribosomes. Top panel indicates abundance of the ncRNAs in total lysate. d , Conventional RIP-qRT-PCR experiments to compare Rmrp association with DDX5 in Th17 and developing thymocytes. Results are representative of three independent experiments. Each experiment was performed with two technical replicates. Graph shows mean ± s.d. ** p

    Article Snippet: Isolated RNA was used in qRT-PCR analysis (Stratagene) to quantify enrichment of RMRP and depletion of other cellular RNAs.

    Techniques: Transduction, Mutagenesis, Expressing, Immunofluorescence, Sequencing, Immunoprecipitation, Quantitative RT-PCR

    Association of Rmrp lncRNA with DDX5 and RORγt in vitro a , In vitro translated HA-tagged wildtype or helicase-dead D DX5 and FLAG-tagged RORγt were incubated with in vitro transcribed Rmrp. After capture on anti-HA or anti-FLAG beads, the amount of lncRNA was determined by qRT-PCR. Data are representative of two independent experiments. Each experiment was performed with two technical replicates. Graphs show mean ± s.d. *** p

    Journal: Nature

    Article Title: DDX5 and its associated lncRNA Rmrp modulate Th17 cell effector functions

    doi: 10.1038/nature16193

    Figure Lengend Snippet: Association of Rmrp lncRNA with DDX5 and RORγt in vitro a , In vitro translated HA-tagged wildtype or helicase-dead D DX5 and FLAG-tagged RORγt were incubated with in vitro transcribed Rmrp. After capture on anti-HA or anti-FLAG beads, the amount of lncRNA was determined by qRT-PCR. Data are representative of two independent experiments. Each experiment was performed with two technical replicates. Graphs show mean ± s.d. *** p

    Article Snippet: Isolated RNA was used in qRT-PCR analysis (Stratagene) to quantify enrichment of RMRP and depletion of other cellular RNAs.

    Techniques: In Vitro, Incubation, Quantitative RT-PCR

    ADAMTS5 and IGFBP5 are downregulated by miR-140 in CRC cells. a The mRNAs of ADAMTS5 and IGFBP5 contain putative binding sites of miR-140. b HCT116 and RKO cells were transfected with miR-140 mimic using oligofectamine. Oligofectamine alone (Control) and negative miRNA (NC) were the negative controls. The relative expression of miR-140 was determined by qRT-PCR after normalization to the internal control, RNU6B. * P

    Journal: Stem Cell Research & Therapy

    Article Title: microRNA -140-5p inhibits colorectal cancer invasion and metastasis by targeting ADAMTS5 and IGFBP5

    doi: 10.1186/s13287-016-0438-5

    Figure Lengend Snippet: ADAMTS5 and IGFBP5 are downregulated by miR-140 in CRC cells. a The mRNAs of ADAMTS5 and IGFBP5 contain putative binding sites of miR-140. b HCT116 and RKO cells were transfected with miR-140 mimic using oligofectamine. Oligofectamine alone (Control) and negative miRNA (NC) were the negative controls. The relative expression of miR-140 was determined by qRT-PCR after normalization to the internal control, RNU6B. * P

    Article Snippet: For qRT-PCR analysis of mRNA expression, cDNA was synthesized using the PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Dalian, China).

    Techniques: Binding Assay, Transfection, Expressing, Quantitative RT-PCR

    ADAMTS5 and IGFBP5 are inversely correlated with the expression of miR-140 and positively correlated with the tumor stage and metastasis of CRC. a qRT-PCR analysis of ADAMTS5 and IGFBP5 expressions in 60 pairs of CRC and adjacent nontumorous tissues. b Correlation of ADAMTS5 and IGFBP5 and miR-140 expression was analyzed by Spearman correlation test in CRC tissues. c Immunohistochemistry analysis of ADAMTS5 and IGFBP5 expressions in CRC tissues. 1, 2, 5 and 6: magnification × 20; and 3, 4, 7 and 8: magnification × 40; 1, 3, 5 and 7: primary sites of tumor tissue; and 2, 4, 6 and 8: noncancerous region of CRC. Scale bar = 100 μm. The results of immunohistochemistry were evaluated by the staining scores. * P

    Journal: Stem Cell Research & Therapy

    Article Title: microRNA -140-5p inhibits colorectal cancer invasion and metastasis by targeting ADAMTS5 and IGFBP5

    doi: 10.1186/s13287-016-0438-5

    Figure Lengend Snippet: ADAMTS5 and IGFBP5 are inversely correlated with the expression of miR-140 and positively correlated with the tumor stage and metastasis of CRC. a qRT-PCR analysis of ADAMTS5 and IGFBP5 expressions in 60 pairs of CRC and adjacent nontumorous tissues. b Correlation of ADAMTS5 and IGFBP5 and miR-140 expression was analyzed by Spearman correlation test in CRC tissues. c Immunohistochemistry analysis of ADAMTS5 and IGFBP5 expressions in CRC tissues. 1, 2, 5 and 6: magnification × 20; and 3, 4, 7 and 8: magnification × 40; 1, 3, 5 and 7: primary sites of tumor tissue; and 2, 4, 6 and 8: noncancerous region of CRC. Scale bar = 100 μm. The results of immunohistochemistry were evaluated by the staining scores. * P

    Article Snippet: For qRT-PCR analysis of mRNA expression, cDNA was synthesized using the PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Dalian, China).

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

    miR-140 is decreased in CRC specimens. The qRT-PCR analysis of miR-140 was performed in 60 paired human CRC and the matched adjacent noncancerous tissues. Expression level of miR-140 was normalized by the internal control RNU6B in each sample. * P

    Journal: Stem Cell Research & Therapy

    Article Title: microRNA -140-5p inhibits colorectal cancer invasion and metastasis by targeting ADAMTS5 and IGFBP5

    doi: 10.1186/s13287-016-0438-5

    Figure Lengend Snippet: miR-140 is decreased in CRC specimens. The qRT-PCR analysis of miR-140 was performed in 60 paired human CRC and the matched adjacent noncancerous tissues. Expression level of miR-140 was normalized by the internal control RNU6B in each sample. * P

    Article Snippet: For qRT-PCR analysis of mRNA expression, cDNA was synthesized using the PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa, Dalian, China).

    Techniques: Quantitative RT-PCR, Expressing

    PolyIC/dsRBEC induces the expression and secretion of pro-inflammatory cytokines in MDA-MB-468 cells. A) qRT-PCR analysis of IFN-β, CCL5, IP10 and TNFα mRNA expression following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 2 and 4 hours. Data were normalized to GAPDH and are expressed as fold change relative to vehicle-treated samples. A representative experiment out of 3 experiments is shown. Error bars represent RQ max. B) Protein levels of IFN-β, CCL5, IP10 and TNFα were measured by ELISA following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 24 hours. Values are averages of triplicate biological samples from one representative experiment. (***,P

    Journal: PLoS ONE

    Article Title: Targeting polyIC to EGFR over-expressing cells using a dsRNA binding protein domain tethered to EGF

    doi: 10.1371/journal.pone.0162321

    Figure Lengend Snippet: PolyIC/dsRBEC induces the expression and secretion of pro-inflammatory cytokines in MDA-MB-468 cells. A) qRT-PCR analysis of IFN-β, CCL5, IP10 and TNFα mRNA expression following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 2 and 4 hours. Data were normalized to GAPDH and are expressed as fold change relative to vehicle-treated samples. A representative experiment out of 3 experiments is shown. Error bars represent RQ max. B) Protein levels of IFN-β, CCL5, IP10 and TNFα were measured by ELISA following treatment with dsRBEC alone, polyIC alone or polyIC/dsRBEC for 24 hours. Values are averages of triplicate biological samples from one representative experiment. (***,P

    Article Snippet: 1 μg of total RNA was reverse-transcribed using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems) and the resulting cDNA was used for qRT-PCR analysis (Fast SYBR Green; Applied Biosystems) using the primer pairs listed in .

    Techniques: Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    Macrophages are recruited to the myocardium in lipotoxic cardiomyopathy. A : hearts from 4-wk-old ACS and NTG littermates were harvested and qRT-PCR was performed on total mRNA to assess the expression of macrophage markers. Graphs show mean expression

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Macrophages modulate cardiac function in lipotoxic cardiomyopathy

    doi: 10.1152/ajpheart.00111.2012

    Figure Lengend Snippet: Macrophages are recruited to the myocardium in lipotoxic cardiomyopathy. A : hearts from 4-wk-old ACS and NTG littermates were harvested and qRT-PCR was performed on total mRNA to assess the expression of macrophage markers. Graphs show mean expression

    Article Snippet: For the qRT-PCR array (SA Biosciences; PAMM-011A), RNA samples were further purified after initial RNazol B/chloroform using RNaeasy columns (Qiagen).

    Techniques: Quantitative RT-PCR, Expressing

    Spatial and temporal distribution of VvCCD1 , VvCCD4a and VvCCD4b transcripts in distinct grapevine tissues. qRT-PCR analysis of VvCCD1 , VvCCD4a and VvCCD4b in leaf, flower and three berry developmental stages. Data are expressed relative to pre-véraison berry stage and normalised to the housekeeping gene, VvEF1a . Relative changes in the total carotenoid and chlorophyll concentrations in the respective tissues are also shown.

    Journal: BMC Plant Biology

    Article Title: Functional characterisation of three members of the Vitis vinifera L. carotenoid cleavage dioxygenase gene family

    doi: 10.1186/1471-2229-13-156

    Figure Lengend Snippet: Spatial and temporal distribution of VvCCD1 , VvCCD4a and VvCCD4b transcripts in distinct grapevine tissues. qRT-PCR analysis of VvCCD1 , VvCCD4a and VvCCD4b in leaf, flower and three berry developmental stages. Data are expressed relative to pre-véraison berry stage and normalised to the housekeeping gene, VvEF1a . Relative changes in the total carotenoid and chlorophyll concentrations in the respective tissues are also shown.

    Article Snippet: KAPA SYBR® FAST qRT-PCR Kit was used according to the manufacturer’s (Kapa Biosystems, Cape Town, South Africa) instructions.

    Techniques: Quantitative RT-PCR

    qRT-PCR analysis of VvCCD1 expression in the transgenic grapevine population. (A) Expression of the native/endogenous (dark grey square symbol) and transgenic (light grey square symbol) VvCCD1 in lines transformed with the overexpression cassette (CCD1) (n = 3). (B) Expression of VvCCD1 in lines transformed with the silencing cassette (RNAi) (n = 3). Data are expressed relative to the wild-type (WT) expression and normalised to VvGAPDH expression.

    Journal: BMC Plant Biology

    Article Title: Functional characterisation of three members of the Vitis vinifera L. carotenoid cleavage dioxygenase gene family

    doi: 10.1186/1471-2229-13-156

    Figure Lengend Snippet: qRT-PCR analysis of VvCCD1 expression in the transgenic grapevine population. (A) Expression of the native/endogenous (dark grey square symbol) and transgenic (light grey square symbol) VvCCD1 in lines transformed with the overexpression cassette (CCD1) (n = 3). (B) Expression of VvCCD1 in lines transformed with the silencing cassette (RNAi) (n = 3). Data are expressed relative to the wild-type (WT) expression and normalised to VvGAPDH expression.

    Article Snippet: KAPA SYBR® FAST qRT-PCR Kit was used according to the manufacturer’s (Kapa Biosystems, Cape Town, South Africa) instructions.

    Techniques: Quantitative RT-PCR, Expressing, Transgenic Assay, Transformation Assay, Over Expression

    Th2 cytokines suppress pro-caspase-1 without affecting levels of pro-IL-1β. (A) Taqman quantitative RT-PCR results showing mRNA expression of pro-IL-1β in THP-1s exposed to 100 μg/mL MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. B) Taqman qRT-PCR results showing mRNA levels of pro-caspase-1 in THP-1s exposed to MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. Statistical analysis was performed using a one-way ANOVA with a post hoc Tukey. ***P

    Journal: PLoS ONE

    Article Title: An Allergic Lung Microenvironment Suppresses Carbon Nanotube-Induced Inflammasome Activation via STAT6-Dependent Inhibition of Caspase-1

    doi: 10.1371/journal.pone.0128888

    Figure Lengend Snippet: Th2 cytokines suppress pro-caspase-1 without affecting levels of pro-IL-1β. (A) Taqman quantitative RT-PCR results showing mRNA expression of pro-IL-1β in THP-1s exposed to 100 μg/mL MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. B) Taqman qRT-PCR results showing mRNA levels of pro-caspase-1 in THP-1s exposed to MWCNTs with IL-4, IL-13, or IL-4 and IL-13 co-treatment. Statistical analysis was performed using a one-way ANOVA with a post hoc Tukey. ***P

    Article Snippet: All qRT-PCR primers were purchased from Life Technologies.

    Techniques: Quantitative RT-PCR, Expressing

    Overexpression of CD86FLm provides a survival advantage against silencing of CD86 and CD28 but not against silencing of IRF4. (A) Representative histograms showing the levels of surface CD86 in 3 different cell lines when either CD28, CD86, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are silenced. Thin black histograms at left are unstained pLKO.1-infected controls. (B) Cell death measured at day 4 postinfection via annexin V staining, shown as percentage of pLKO.1-infected controls. (A-B) Data shown are from day 4 postinfection and representative of at least 3 independent experiments. (C) qRT-PCR was performed to determine levels of CD86 , CD28 , and IRF4 to compare the different cell lines. Data are normalized to β-actin as endogenous control and then compared relative to mRNA levels in pLKO.1 empty vector control. Data shown are mean ± SEM of at least 3 independent experiments. (D) Representative western blots showing levels of IRF4 in cells overexpressing CD86FLm or CD86TLm when either CD86 or CD28 are silenced. Lysates are from day 4 postinfection. (E) Cell death as measured by annexin V staining at day 4 postinfection in 8226-pCDNA3.1 or 8226-CD86FLm cells where CD86 or IRF4 was silenced; shown as percentage of pLKO.1-infected controls. (F) Representative western blots showing levels of IRF4 and β-actin in lysates from experiments in panel E. Data shown are mean ± SEM of at least 3 independent experiments. * P

    Journal: Blood Advances

    Article Title: CD86 regulates myeloma cell survival

    doi: 10.1182/bloodadvances.2017011601

    Figure Lengend Snippet: Overexpression of CD86FLm provides a survival advantage against silencing of CD86 and CD28 but not against silencing of IRF4. (A) Representative histograms showing the levels of surface CD86 in 3 different cell lines when either CD28, CD86, or glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are silenced. Thin black histograms at left are unstained pLKO.1-infected controls. (B) Cell death measured at day 4 postinfection via annexin V staining, shown as percentage of pLKO.1-infected controls. (A-B) Data shown are from day 4 postinfection and representative of at least 3 independent experiments. (C) qRT-PCR was performed to determine levels of CD86 , CD28 , and IRF4 to compare the different cell lines. Data are normalized to β-actin as endogenous control and then compared relative to mRNA levels in pLKO.1 empty vector control. Data shown are mean ± SEM of at least 3 independent experiments. (D) Representative western blots showing levels of IRF4 in cells overexpressing CD86FLm or CD86TLm when either CD86 or CD28 are silenced. Lysates are from day 4 postinfection. (E) Cell death as measured by annexin V staining at day 4 postinfection in 8226-pCDNA3.1 or 8226-CD86FLm cells where CD86 or IRF4 was silenced; shown as percentage of pLKO.1-infected controls. (F) Representative western blots showing levels of IRF4 and β-actin in lysates from experiments in panel E. Data shown are mean ± SEM of at least 3 independent experiments. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase and β-actin were used as endogenous control genes. qRT-PCR probes are from Applied Biosystems and are listed in supplemental Table 4.

    Techniques: Over Expression, Infection, Staining, Quantitative RT-PCR, Plasmid Preparation, Western Blot

    Gene expression changes in CD28- vs CD86-silenced cells are consistent with regulation of both distinct and common pathways, including expression of IRF4. (A) Gene set enrichment analysis showing upregulated (top) and downregulated (bottom) gene sets in shCD28- (blue) and shCD86-treated (red) myeloma cells. For each gene set, the enrichment score is shown above the ranked change in gene expression, where genes that overlap the gene set are denoted by blue and red ticks for shCD28 and shCD86, respectively. (B) qRT-PCR showing IRF4 mRNA levels when CD28 or CD86 was silenced in MM.1s or 8226 cells. (C) Representative western blots showing IRF4 levels with silencing of CD28 or CD86 in cell lines indicated at different time points. (D) Integrin levels were measured via qRT-PCR ( ITGB7 and ITGB1 ; left). Representative histograms showing ITGβ1 and ITGβ7 on day 4 after shRNA treatment (right). For qRT-PCR, all data are normalized to β-actin as an endogenous control and then compared relative to mRNA levels in pLKO.1 empty vector control. RNA was extracted at day 3 postinfection. Data shown are mean ± SEM of at least 3 independent experiments. Histograms showing surface levels of indicated molecules. Gray histograms at left represent unstained or isotype controls. Flow cytometry data shown are from day 4 postinfection with lentiviral vectors. Flow cytometry and western blot data shown are representative of at least 3 independent experiments. (E) Cell adhesion 3 days postinfection with lentivirus containing the indicated shRNA. Myeloma cells stained with calcein AM were cocultured with HS-5 cells for 2 hours. Fluorescence was measured (485/528 emission/excitation) with BioTek Synergy H1 multiwell plate reader, and data are presented as fluorescence relative to pLKO.1 controls (mean ± SEM). Prewash readings were taken to ensure similar numbers of live cells were added to each well. All samples were plated in triplicate. Data are mean of 3 independent experiments. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GX, gene expression changes; RFU, relative fluorescence unit.

    Journal: Blood Advances

    Article Title: CD86 regulates myeloma cell survival

    doi: 10.1182/bloodadvances.2017011601

    Figure Lengend Snippet: Gene expression changes in CD28- vs CD86-silenced cells are consistent with regulation of both distinct and common pathways, including expression of IRF4. (A) Gene set enrichment analysis showing upregulated (top) and downregulated (bottom) gene sets in shCD28- (blue) and shCD86-treated (red) myeloma cells. For each gene set, the enrichment score is shown above the ranked change in gene expression, where genes that overlap the gene set are denoted by blue and red ticks for shCD28 and shCD86, respectively. (B) qRT-PCR showing IRF4 mRNA levels when CD28 or CD86 was silenced in MM.1s or 8226 cells. (C) Representative western blots showing IRF4 levels with silencing of CD28 or CD86 in cell lines indicated at different time points. (D) Integrin levels were measured via qRT-PCR ( ITGB7 and ITGB1 ; left). Representative histograms showing ITGβ1 and ITGβ7 on day 4 after shRNA treatment (right). For qRT-PCR, all data are normalized to β-actin as an endogenous control and then compared relative to mRNA levels in pLKO.1 empty vector control. RNA was extracted at day 3 postinfection. Data shown are mean ± SEM of at least 3 independent experiments. Histograms showing surface levels of indicated molecules. Gray histograms at left represent unstained or isotype controls. Flow cytometry data shown are from day 4 postinfection with lentiviral vectors. Flow cytometry and western blot data shown are representative of at least 3 independent experiments. (E) Cell adhesion 3 days postinfection with lentivirus containing the indicated shRNA. Myeloma cells stained with calcein AM were cocultured with HS-5 cells for 2 hours. Fluorescence was measured (485/528 emission/excitation) with BioTek Synergy H1 multiwell plate reader, and data are presented as fluorescence relative to pLKO.1 controls (mean ± SEM). Prewash readings were taken to ensure similar numbers of live cells were added to each well. All samples were plated in triplicate. Data are mean of 3 independent experiments. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GX, gene expression changes; RFU, relative fluorescence unit.

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase and β-actin were used as endogenous control genes. qRT-PCR probes are from Applied Biosystems and are listed in supplemental Table 4.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, shRNA, Plasmid Preparation, Flow Cytometry, Cytometry, Staining, Fluorescence

    Silencing or blockade of CD86 results in myeloma cell death. (A) Myeloma cell lines were infected with lentiviral particles carrying empty vector (pLKO.1) or the individual shRNAs, and cell death was monitored by annexin V–fluorescein isothiocyanate staining for 4 days. Data for different time points were all compared with pLKO.1 controls (left). mRNA quantification as measured via qRT-PCR comparing levels of CD28 or CD86 to vector-controls (middle). Representative histograms show CD28 or CD86 surface levels at day 4 postinfection. Thin gray histograms at left are isotype controls (right). (B) mRNA quantification as measured via qRT-PCR comparing levels of CD28 or CD86 with vector controls (left). Representative histograms show CD28 or CD86 surface expression at day 4 postinfection. Thin gray histograms at left are isotype controls (right). (A-B) All data are presented as mean ± standard error of the mean (SEM) of at least 3 independent experiments. * P

    Journal: Blood Advances

    Article Title: CD86 regulates myeloma cell survival

    doi: 10.1182/bloodadvances.2017011601

    Figure Lengend Snippet: Silencing or blockade of CD86 results in myeloma cell death. (A) Myeloma cell lines were infected with lentiviral particles carrying empty vector (pLKO.1) or the individual shRNAs, and cell death was monitored by annexin V–fluorescein isothiocyanate staining for 4 days. Data for different time points were all compared with pLKO.1 controls (left). mRNA quantification as measured via qRT-PCR comparing levels of CD28 or CD86 to vector-controls (middle). Representative histograms show CD28 or CD86 surface levels at day 4 postinfection. Thin gray histograms at left are isotype controls (right). (B) mRNA quantification as measured via qRT-PCR comparing levels of CD28 or CD86 with vector controls (left). Representative histograms show CD28 or CD86 surface expression at day 4 postinfection. Thin gray histograms at left are isotype controls (right). (A-B) All data are presented as mean ± standard error of the mean (SEM) of at least 3 independent experiments. * P

    Article Snippet: Glyceraldehyde-3-phosphate dehydrogenase and β-actin were used as endogenous control genes. qRT-PCR probes are from Applied Biosystems and are listed in supplemental Table 4.

    Techniques: Infection, Plasmid Preparation, Staining, Quantitative RT-PCR, Expressing