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  • 88
    Jena Bioscience biotin pcr labeling mix
    Biotin Pcr Labeling Mix, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore biotin rna labeling mix
    Western blot analysis of G6PD expression in melanoma cells, and detection of physical interaction between <t>GAS5</t> and G6PD protein using <t>RNA</t> immunoprecipitation (RIP) and RNA pull down assay. a A375, A375-GAS5si, SK-Mel-110, and SK-Mel-110-GAS5over cell lysates were subjected to ( a , left) western blot analysis to measure G6PD protein expression with western blotting results, and ( a , right) the results are expressed in histogram format as the ratio of target band density to that of the GAPDH loading control (mean ± standard deviation) for three independent experiments (* P
    Biotin Rna Labeling Mix, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche biotin rna labeling mix
    AVAN direct binds to TRIM25 and enhances the antivirus immune response (A) <t>RNA</t> pull-down of AVAN -associated proteins using biotinylated AVAN or <t>antisense</t> probes. Isolated proteins were resolved by SDS-PAGE followed by silver staining. (B) Pull-down western blot showing that AVAN can bind directly to TRIM25. (C) ChIRP followed by western blot show that AVAN can bind to TRIM25. (D and E) Exogenous (D) and endogenous (E) RIP of TRIM25 in BJ501 infected cells using anti-TRIM25 or anti-IgG antibodies. The relative enrichment fold of AVAN was calculated by qRT-PCR. (F) AVAN pull-down western blot with lysates of A549 cells transfected with Flag, Flag-TRIM25, Flag-SPRY, Flag-B Box/CCD or Flag-Ring. (G) Truncated AVAN pull-down, truncates (upper panel) were obtained via in vitro transcription and incubated with BJ501-infected A549 lysates for RNA pulldown. (H and I) TRIM25 co-immunoprecipitation with proteins from lysates of BJ501-infected A549 cells transfected with AVAN s or siRNAs, followed by immunoblotting. Anti-TRIM25 and anti-RIG-I antibodies were used for immunoprecipitated. (J and K) Immunoblot analysis of endogenous RIG-I ubiquitylation in BJ501-infected A549 cells transfected with AVAN s or siRNAs. Anti-RIG-I antibody was used for immunoprecipitated. (L) Immunoblot analysis of proteins immunoprecipitated with anti-Flag from lysates of BJ501-infected A549 cells transfected with AVAN , HA-Ub and Flag-tagged RIG-I. (M and N) IFN-alpha (M) and IFN-beta (N) expression upon AVAN transfection in A549 cells that were infected by BJ501 or not (MOI=1) at 24h post-infection, and then individually knock down RIG-I or TRIM25, analyzed by qRT-PCR (n=3; means ± SEM; *p
    Biotin Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 2514 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher biotin rna labeling mix
    Regulatory Mechanism of lncITPF in Its Host Gene (A) Itgbl1 expression was higher in fibrotic tissue than in normal tissue on the basis of <t>RNA</t> sequencing. (B) Itgbl1 was upregulated in cells treated with 5 ng/mL TGF-β1 for 12, 24, 48, and 72 hr. (C) qRT-PCR analysis showed that Itgbl1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (D) Western blot showed that ITGBL1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (E) Single-molecule RNA-FISH detecting the location of lncITPF (red) in cells. U6 and 18S RNA were used as cytoplasmic and nuclear localization markers, respectively. DNA (blue) was stained with DAPI. A representative image is shown. (F) qRT-PCR analysis of <t>RNAs</t> purified from nucleoplasmic (red), chromatin nuclear (gray), and cytosolic (blue) compartments in cells. NC indicates a negative control, BP indicates blank plasmid, and RP indicates the recombinant plasmid of the overexpressed lncITPF. Each bar represents the mean ± SD; n = 6; **p
    Biotin Rna Labeling Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GenePharma Company biotin rna labeling mix
    ELK4 expression in Aβ 1-42 -incubated <t>ECs</t> and ELK4 regulated BBB permeability in AD microenvironment. (A) <t>RNA</t> microarray analysis was performed in ECs treated with shLINC00662. Red indicates high relative expression and green indicates low relative expression. (B) Relative expression levels of ZEB1, ELK4, and ART1 determined by qRT-PCR. Data represent mean ± SD (n = 3, each). **P
    Biotin Rna Labeling Mix, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche biotin rna labelling mix
    <t>Atrolnc‐1</t> regulates MuRF‐1 via interacting with ABIN‐1 in vitro and in vivo . (A) In situ hybridization analysis with a biotin‐labelled Atrolnc‐1 antisense probe in C2C12 cell. Atrolnc‐1 positive signals manifest as brown granules located both in the nuclear (N) and cytoplasm (Cyto). Atrolnc‐1 sense probe was used as negative control (CTL). Scale bar = 20 μm. (B) SDS‐PAGE gels exhibit the results of <t>RNA</t> pull‐down assay. The interaction of biotin‐labelled Atrolnc‐1 probe (marked as 2) with proteins from cytoplasm (left gel) and nucleus (right gel) extracted from C2C12 cells are shown. Atrolnc‐1 antisense probe was used as control (marked as 1). M represented molecular marker. (C) The table lists mass spectrometry result of proteins interacting with Atrolnc‐1. In the gels in (B), ABIN‐1, an inhibitor of NF‐kB signalling, was found to bind with biotin‐labelled Atrolnc‐1 probe in cytoplasmic extracts from myotubes. The iBAQ denotes the sum of all the peptides intensities divided by the number of observable peptides of a protein. (D) Left panel: a representative immunoblotting shows a band at 72 kD using ABIN‐1 antibody in the proteins pulled‐down by biotin‐labelled Atrolnc‐1 probe. Right panel: in cytoplasmic extracts of tibialis anterior (TA) muscle from mice, immunoblotting also detected a band at 72 kD in RNA pull‐down assay by biotin‐labelled Atrolnc‐1 probe. (E) The result of immunoprecipitation (IP) in TA muscle from mice with or without chronic kidney disease (CKD). Ten per cent of the IP product was used to conduct western blot (low panel), showing a band at 72 kD using ABIN‐1 antibody (ABIN‐1 Ab) while a RT‐PCR of Arolnc‐1(upper panel) was performed using the rest of IP product. Notice that the Atrolnc‐1 interacted with ABIN‐1 was increased in CKD mice. Mock, normal mouse IgG. (F) Immunoblot in upper panel shows increased ABIN‐1 expression in C2C12 myotubes after transduced by CRISPR/dCas9 mediated synergistic activation mediator transcription activation system. Bar graph (low panel) illustrates the quantity result (mean ± SEM; n = 5, * P
    Biotin Rna Labelling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 88/100, based on 203 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim biotin rna labelling mix
    <t>Atrolnc‐1</t> regulates MuRF‐1 via interacting with ABIN‐1 in vitro and in vivo . (A) In situ hybridization analysis with a biotin‐labelled Atrolnc‐1 antisense probe in C2C12 cell. Atrolnc‐1 positive signals manifest as brown granules located both in the nuclear (N) and cytoplasm (Cyto). Atrolnc‐1 sense probe was used as negative control (CTL). Scale bar = 20 μm. (B) SDS‐PAGE gels exhibit the results of <t>RNA</t> pull‐down assay. The interaction of biotin‐labelled Atrolnc‐1 probe (marked as 2) with proteins from cytoplasm (left gel) and nucleus (right gel) extracted from C2C12 cells are shown. Atrolnc‐1 antisense probe was used as control (marked as 1). M represented molecular marker. (C) The table lists mass spectrometry result of proteins interacting with Atrolnc‐1. In the gels in (B), ABIN‐1, an inhibitor of NF‐kB signalling, was found to bind with biotin‐labelled Atrolnc‐1 probe in cytoplasmic extracts from myotubes. The iBAQ denotes the sum of all the peptides intensities divided by the number of observable peptides of a protein. (D) Left panel: a representative immunoblotting shows a band at 72 kD using ABIN‐1 antibody in the proteins pulled‐down by biotin‐labelled Atrolnc‐1 probe. Right panel: in cytoplasmic extracts of tibialis anterior (TA) muscle from mice, immunoblotting also detected a band at 72 kD in RNA pull‐down assay by biotin‐labelled Atrolnc‐1 probe. (E) The result of immunoprecipitation (IP) in TA muscle from mice with or without chronic kidney disease (CKD). Ten per cent of the IP product was used to conduct western blot (low panel), showing a band at 72 kD using ABIN‐1 antibody (ABIN‐1 Ab) while a RT‐PCR of Arolnc‐1(upper panel) was performed using the rest of IP product. Notice that the Atrolnc‐1 interacted with ABIN‐1 was increased in CKD mice. Mock, normal mouse IgG. (F) Immunoblot in upper panel shows increased ABIN‐1 expression in C2C12 myotubes after transduced by CRISPR/dCas9 mediated synergistic activation mediator transcription activation system. Bar graph (low panel) illustrates the quantity result (mean ± SEM; n = 5, * P
    Biotin Rna Labelling Mix, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche biotin rna labeling mix sp6 t7
    <t>Atrolnc‐1</t> regulates MuRF‐1 via interacting with ABIN‐1 in vitro and in vivo . (A) In situ hybridization analysis with a biotin‐labelled Atrolnc‐1 antisense probe in C2C12 cell. Atrolnc‐1 positive signals manifest as brown granules located both in the nuclear (N) and cytoplasm (Cyto). Atrolnc‐1 sense probe was used as negative control (CTL). Scale bar = 20 μm. (B) SDS‐PAGE gels exhibit the results of <t>RNA</t> pull‐down assay. The interaction of biotin‐labelled Atrolnc‐1 probe (marked as 2) with proteins from cytoplasm (left gel) and nucleus (right gel) extracted from C2C12 cells are shown. Atrolnc‐1 antisense probe was used as control (marked as 1). M represented molecular marker. (C) The table lists mass spectrometry result of proteins interacting with Atrolnc‐1. In the gels in (B), ABIN‐1, an inhibitor of NF‐kB signalling, was found to bind with biotin‐labelled Atrolnc‐1 probe in cytoplasmic extracts from myotubes. The iBAQ denotes the sum of all the peptides intensities divided by the number of observable peptides of a protein. (D) Left panel: a representative immunoblotting shows a band at 72 kD using ABIN‐1 antibody in the proteins pulled‐down by biotin‐labelled Atrolnc‐1 probe. Right panel: in cytoplasmic extracts of tibialis anterior (TA) muscle from mice, immunoblotting also detected a band at 72 kD in RNA pull‐down assay by biotin‐labelled Atrolnc‐1 probe. (E) The result of immunoprecipitation (IP) in TA muscle from mice with or without chronic kidney disease (CKD). Ten per cent of the IP product was used to conduct western blot (low panel), showing a band at 72 kD using ABIN‐1 antibody (ABIN‐1 Ab) while a RT‐PCR of Arolnc‐1(upper panel) was performed using the rest of IP product. Notice that the Atrolnc‐1 interacted with ABIN‐1 was increased in CKD mice. Mock, normal mouse IgG. (F) Immunoblot in upper panel shows increased ABIN‐1 expression in C2C12 myotubes after transduced by CRISPR/dCas9 mediated synergistic activation mediator transcription activation system. Bar graph (low panel) illustrates the quantity result (mean ± SEM; n = 5, * P
    Biotin Rna Labeling Mix Sp6 T7, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim biotin rna labeling mix
    In situ hybridization of rice root apices with probes specific for transcripts of cdc2Os1 , cdc2Os2 , cdc2Os3 , and R2 . Longitudinal sections of roots were allowed to hybridize with <t>digoxigenin-labeled</t> <t>RNA</t> probes. Hybridization signals are visible as brownish-purple staining. a, cdc2Os1 antisense probe; b, cdc2Os2 antisense probe; c, cdc2Os3 antisense probe; d, R2 antisense probe; and e, cdc2Os1 sense probe. Bar = 100 μm.
    Biotin Rna Labeling Mix, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche biotin rna labeling mix kit
    <t>EPIC1</t> is a nuclear lncRNA regulating MYC targets expression (A) qRT-PCR analysis of EPIC1 expression (top) and Western blot (bottom) of subcellular fractionation in MCF-7cells. GAPDH and U6 <t>RNA</t> served as a marker for cytoplasmic and nuclear gene localization, respectively. SNRP70 and GAPDH served as a specific nuclear and cytoplasmic marker to whole cell lysates (WCL), cytoplasmic (Cyto), and nuclear fractionation (Nuc). Error bars indicate mean ± SD, n = 3 for technical replicates. (B) Schematic of the identification of EPIC1 correlated genes in breast tumors from TCGA (yellow), and genes potentially regulated by EPIC1 in MCF-7 cells (green). (C) Co-expression analysis showing that EPIC1 expression is associated with 2005 genes in 559 patients with breast cancer (BRCA). Each column represents one patient. (D) GSEA analysis of the EPIC1 -related pathways in 20 cancer types (left panel) and EPIC1 knockdown MCF-7 cells (right panel). The heatmap indicates the GSEA scores. (E) Association between the enrichment of MYC targets and EPIC1 expression in breast tumors by GSEA analysis (D). (F) EPIC1 -regulated gene expression by qRT-PCR analysis (top) and RNA-seq (bottom). Error bars indicate mean ± SD, n = 3 for technical replicates. (G) Western blot of MYC-regulated targets in MCF-7 (left) and ZR-75-1 (right) cells treated with EPIC1 and MYC siRNAs. .
    Biotin Rna Labeling Mix Kit, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche spiked in biotin rna labeling mix
    <t>EPIC1</t> is a nuclear lncRNA regulating MYC targets expression (A) qRT-PCR analysis of EPIC1 expression (top) and Western blot (bottom) of subcellular fractionation in MCF-7cells. GAPDH and U6 <t>RNA</t> served as a marker for cytoplasmic and nuclear gene localization, respectively. SNRP70 and GAPDH served as a specific nuclear and cytoplasmic marker to whole cell lysates (WCL), cytoplasmic (Cyto), and nuclear fractionation (Nuc). Error bars indicate mean ± SD, n = 3 for technical replicates. (B) Schematic of the identification of EPIC1 correlated genes in breast tumors from TCGA (yellow), and genes potentially regulated by EPIC1 in MCF-7 cells (green). (C) Co-expression analysis showing that EPIC1 expression is associated with 2005 genes in 559 patients with breast cancer (BRCA). Each column represents one patient. (D) GSEA analysis of the EPIC1 -related pathways in 20 cancer types (left panel) and EPIC1 knockdown MCF-7 cells (right panel). The heatmap indicates the GSEA scores. (E) Association between the enrichment of MYC targets and EPIC1 expression in breast tumors by GSEA analysis (D). (F) EPIC1 -regulated gene expression by qRT-PCR analysis (top) and RNA-seq (bottom). Error bars indicate mean ± SD, n = 3 for technical replicates. (G) Western blot of MYC-regulated targets in MCF-7 (left) and ZR-75-1 (right) cells treated with EPIC1 and MYC siRNAs. .
    Spiked In Biotin Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim biotin rna labeling mix kit
    <t>EPIC1</t> is a nuclear lncRNA regulating MYC targets expression (A) qRT-PCR analysis of EPIC1 expression (top) and Western blot (bottom) of subcellular fractionation in MCF-7cells. GAPDH and U6 <t>RNA</t> served as a marker for cytoplasmic and nuclear gene localization, respectively. SNRP70 and GAPDH served as a specific nuclear and cytoplasmic marker to whole cell lysates (WCL), cytoplasmic (Cyto), and nuclear fractionation (Nuc). Error bars indicate mean ± SD, n = 3 for technical replicates. (B) Schematic of the identification of EPIC1 correlated genes in breast tumors from TCGA (yellow), and genes potentially regulated by EPIC1 in MCF-7 cells (green). (C) Co-expression analysis showing that EPIC1 expression is associated with 2005 genes in 559 patients with breast cancer (BRCA). Each column represents one patient. (D) GSEA analysis of the EPIC1 -related pathways in 20 cancer types (left panel) and EPIC1 knockdown MCF-7 cells (right panel). The heatmap indicates the GSEA scores. (E) Association between the enrichment of MYC targets and EPIC1 expression in breast tumors by GSEA analysis (D). (F) EPIC1 -regulated gene expression by qRT-PCR analysis (top) and RNA-seq (bottom). Error bars indicate mean ± SD, n = 3 for technical replicates. (G) Western blot of MYC-regulated targets in MCF-7 (left) and ZR-75-1 (right) cells treated with EPIC1 and MYC siRNAs. .
    Biotin Rna Labeling Mix Kit, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche biotinylated rna labeling mix
    EGFR-AS1 promotes the maintenance of EGFR mRNA stability by binding to HuR. a Silver staining SDS-PAGE gel of electrophoretically separated proteins immunoprecipitated with EGFR-AS1 and its antisense <t>RNA</t> in 786-O cells. b RNA pull-down assay was performed in 786-O and A498 cells using <t>biotinylated</t> EGFR-AS1 or antisense RNA probe transcribed in vitro and detected by western blots. c Upper: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR-AS1 RNA enrichment in immunoprecipitated complexes. IgG is the negative control. Lower: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR RNA enrichment in immunoprecipitated complexes. d RIP assay of the enrichment of EGFR mRNA with HuR between the EGFR-AS1 knockdown and NC group in RCC cells. IgG was used as an internal control. e Upper: RNA FISH analysis of EGFR mRNA (green) and immunofluorescence detection of HuR (red) in RCC cells. The rightmost graph shows colocalization between the green signal (EGFR) and the red signal (HuR). Pearson’s R = 0.583. Scale bar = 50 μm. Lower: RNA FISH analysis of EGFR-AS1 (green) and immunofluorescence detection of HuR (red) in RCC cells. Pearson’s R = 0.416. f The rate of degradation of the EGFR mRNA between the HuR knockdown and control group using RNA stability assays in RCC cells. g The rate of degradation of the EGFR mRNA between the HuR overexpressing and control group using RNA stability assays in RCC cells. h The rate of degradation of the EGFR mRNA in the EGFR-AS1 knockdown and control cells transfected with pcDNA3.1 + -HuR over 12 h in KETR-3 and ACHN cells. i The rate of degradation of the EGFR mRNA in the EGFR-AS1 overexpressing and control cells transfected with HuR siRNA over 12 h
    Biotinylated Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche dig rna labeling mix
    EGFR-AS1 promotes the maintenance of EGFR mRNA stability by binding to HuR. a Silver staining SDS-PAGE gel of electrophoretically separated proteins immunoprecipitated with EGFR-AS1 and its antisense <t>RNA</t> in 786-O cells. b RNA pull-down assay was performed in 786-O and A498 cells using <t>biotinylated</t> EGFR-AS1 or antisense RNA probe transcribed in vitro and detected by western blots. c Upper: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR-AS1 RNA enrichment in immunoprecipitated complexes. IgG is the negative control. Lower: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR RNA enrichment in immunoprecipitated complexes. d RIP assay of the enrichment of EGFR mRNA with HuR between the EGFR-AS1 knockdown and NC group in RCC cells. IgG was used as an internal control. e Upper: RNA FISH analysis of EGFR mRNA (green) and immunofluorescence detection of HuR (red) in RCC cells. The rightmost graph shows colocalization between the green signal (EGFR) and the red signal (HuR). Pearson’s R = 0.583. Scale bar = 50 μm. Lower: RNA FISH analysis of EGFR-AS1 (green) and immunofluorescence detection of HuR (red) in RCC cells. Pearson’s R = 0.416. f The rate of degradation of the EGFR mRNA between the HuR knockdown and control group using RNA stability assays in RCC cells. g The rate of degradation of the EGFR mRNA between the HuR overexpressing and control group using RNA stability assays in RCC cells. h The rate of degradation of the EGFR mRNA in the EGFR-AS1 knockdown and control cells transfected with pcDNA3.1 + -HuR over 12 h in KETR-3 and ACHN cells. i The rate of degradation of the EGFR mRNA in the EGFR-AS1 overexpressing and control cells transfected with HuR siRNA over 12 h
    Dig Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 5197 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche dig rna labelling mixture biotin rna labelling mix
    EGFR-AS1 promotes the maintenance of EGFR mRNA stability by binding to HuR. a Silver staining SDS-PAGE gel of electrophoretically separated proteins immunoprecipitated with EGFR-AS1 and its antisense <t>RNA</t> in 786-O cells. b RNA pull-down assay was performed in 786-O and A498 cells using <t>biotinylated</t> EGFR-AS1 or antisense RNA probe transcribed in vitro and detected by western blots. c Upper: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR-AS1 RNA enrichment in immunoprecipitated complexes. IgG is the negative control. Lower: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR RNA enrichment in immunoprecipitated complexes. d RIP assay of the enrichment of EGFR mRNA with HuR between the EGFR-AS1 knockdown and NC group in RCC cells. IgG was used as an internal control. e Upper: RNA FISH analysis of EGFR mRNA (green) and immunofluorescence detection of HuR (red) in RCC cells. The rightmost graph shows colocalization between the green signal (EGFR) and the red signal (HuR). Pearson’s R = 0.583. Scale bar = 50 μm. Lower: RNA FISH analysis of EGFR-AS1 (green) and immunofluorescence detection of HuR (red) in RCC cells. Pearson’s R = 0.416. f The rate of degradation of the EGFR mRNA between the HuR knockdown and control group using RNA stability assays in RCC cells. g The rate of degradation of the EGFR mRNA between the HuR overexpressing and control group using RNA stability assays in RCC cells. h The rate of degradation of the EGFR mRNA in the EGFR-AS1 knockdown and control cells transfected with pcDNA3.1 + -HuR over 12 h in KETR-3 and ACHN cells. i The rate of degradation of the EGFR mRNA in the EGFR-AS1 overexpressing and control cells transfected with HuR siRNA over 12 h
    Dig Rna Labelling Mixture Biotin Rna Labelling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Genisphere flashtag biotin rna labeling kit
    Number of detected probes and probe sets in hybridizations using <t>RNA</t> from four tumor samples labeled with the <t>FlashTag</t> Biotin- and FlashTag Biotin HSR labeling kits .
    Flashtag Biotin Rna Labeling Kit, supplied by Genisphere, used in various techniques. Bioz Stars score: 92/100, based on 421 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche t7 rna polymerase biotin rna labeling mix
    Number of detected probes and probe sets in hybridizations using <t>RNA</t> from four tumor samples labeled with the <t>FlashTag</t> Biotin- and FlashTag Biotin HSR labeling kits .
    T7 Rna Polymerase Biotin Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher flashtag biotin hsr rna labeling kits
    Number of detected probes and probe sets in hybridizations using <t>RNA</t> from four tumor samples labeled with the <t>FlashTag</t> Biotin- and FlashTag Biotin HSR labeling kits .
    Flashtag Biotin Hsr Rna Labeling Kits, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Western blot analysis of G6PD expression in melanoma cells, and detection of physical interaction between GAS5 and G6PD protein using RNA immunoprecipitation (RIP) and RNA pull down assay. a A375, A375-GAS5si, SK-Mel-110, and SK-Mel-110-GAS5over cell lysates were subjected to ( a , left) western blot analysis to measure G6PD protein expression with western blotting results, and ( a , right) the results are expressed in histogram format as the ratio of target band density to that of the GAPDH loading control (mean ± standard deviation) for three independent experiments (* P

    Journal: Journal of Cancer Research and Clinical Oncology

    Article Title: LncRNA GAS5 regulates redox balance and dysregulates the cell cycle and apoptosis in malignant melanoma cells

    doi: 10.1007/s00432-018-2820-4

    Figure Lengend Snippet: Western blot analysis of G6PD expression in melanoma cells, and detection of physical interaction between GAS5 and G6PD protein using RNA immunoprecipitation (RIP) and RNA pull down assay. a A375, A375-GAS5si, SK-Mel-110, and SK-Mel-110-GAS5over cell lysates were subjected to ( a , left) western blot analysis to measure G6PD protein expression with western blotting results, and ( a , right) the results are expressed in histogram format as the ratio of target band density to that of the GAPDH loading control (mean ± standard deviation) for three independent experiments (* P

    Article Snippet: RNA–protein coimmunoprecipitation LncRNA-GAS5 cDNA was generated by reverse transcription and PCR, and the cDNA was ligated into pGEM-T Easy (Promega, Madison, WI, USA) to generate GAS5-pGEM-T. LncRNA-GAS5 was transcribed in vitro and biotin-labeled using Biotin RNA Labeling Mix (Sigma-Aldrich).

    Techniques: Western Blot, Expressing, Immunoprecipitation, Pull Down Assay, Standard Deviation

    AVAN direct binds to TRIM25 and enhances the antivirus immune response (A) RNA pull-down of AVAN -associated proteins using biotinylated AVAN or antisense probes. Isolated proteins were resolved by SDS-PAGE followed by silver staining. (B) Pull-down western blot showing that AVAN can bind directly to TRIM25. (C) ChIRP followed by western blot show that AVAN can bind to TRIM25. (D and E) Exogenous (D) and endogenous (E) RIP of TRIM25 in BJ501 infected cells using anti-TRIM25 or anti-IgG antibodies. The relative enrichment fold of AVAN was calculated by qRT-PCR. (F) AVAN pull-down western blot with lysates of A549 cells transfected with Flag, Flag-TRIM25, Flag-SPRY, Flag-B Box/CCD or Flag-Ring. (G) Truncated AVAN pull-down, truncates (upper panel) were obtained via in vitro transcription and incubated with BJ501-infected A549 lysates for RNA pulldown. (H and I) TRIM25 co-immunoprecipitation with proteins from lysates of BJ501-infected A549 cells transfected with AVAN s or siRNAs, followed by immunoblotting. Anti-TRIM25 and anti-RIG-I antibodies were used for immunoprecipitated. (J and K) Immunoblot analysis of endogenous RIG-I ubiquitylation in BJ501-infected A549 cells transfected with AVAN s or siRNAs. Anti-RIG-I antibody was used for immunoprecipitated. (L) Immunoblot analysis of proteins immunoprecipitated with anti-Flag from lysates of BJ501-infected A549 cells transfected with AVAN , HA-Ub and Flag-tagged RIG-I. (M and N) IFN-alpha (M) and IFN-beta (N) expression upon AVAN transfection in A549 cells that were infected by BJ501 or not (MOI=1) at 24h post-infection, and then individually knock down RIG-I or TRIM25, analyzed by qRT-PCR (n=3; means ± SEM; *p

    Journal: bioRxiv

    Article Title: Long noncoding RNA AVAN promotes antiviral innate immunity by interacting with TRIM25 and enhancing the transcription of FOXO3a

    doi: 10.1101/623132

    Figure Lengend Snippet: AVAN direct binds to TRIM25 and enhances the antivirus immune response (A) RNA pull-down of AVAN -associated proteins using biotinylated AVAN or antisense probes. Isolated proteins were resolved by SDS-PAGE followed by silver staining. (B) Pull-down western blot showing that AVAN can bind directly to TRIM25. (C) ChIRP followed by western blot show that AVAN can bind to TRIM25. (D and E) Exogenous (D) and endogenous (E) RIP of TRIM25 in BJ501 infected cells using anti-TRIM25 or anti-IgG antibodies. The relative enrichment fold of AVAN was calculated by qRT-PCR. (F) AVAN pull-down western blot with lysates of A549 cells transfected with Flag, Flag-TRIM25, Flag-SPRY, Flag-B Box/CCD or Flag-Ring. (G) Truncated AVAN pull-down, truncates (upper panel) were obtained via in vitro transcription and incubated with BJ501-infected A549 lysates for RNA pulldown. (H and I) TRIM25 co-immunoprecipitation with proteins from lysates of BJ501-infected A549 cells transfected with AVAN s or siRNAs, followed by immunoblotting. Anti-TRIM25 and anti-RIG-I antibodies were used for immunoprecipitated. (J and K) Immunoblot analysis of endogenous RIG-I ubiquitylation in BJ501-infected A549 cells transfected with AVAN s or siRNAs. Anti-RIG-I antibody was used for immunoprecipitated. (L) Immunoblot analysis of proteins immunoprecipitated with anti-Flag from lysates of BJ501-infected A549 cells transfected with AVAN , HA-Ub and Flag-tagged RIG-I. (M and N) IFN-alpha (M) and IFN-beta (N) expression upon AVAN transfection in A549 cells that were infected by BJ501 or not (MOI=1) at 24h post-infection, and then individually knock down RIG-I or TRIM25, analyzed by qRT-PCR (n=3; means ± SEM; *p

    Article Snippet: In vitro biotin-labeled RNAs (lncAVAN and its antisense RNA) were transcribed with biotin RNA labeling mix (Roche) and T7 RNA polymerase (Roche), treated with RNase-free DNase I (Promega) and purified using the RNeasy Mini Kit (QIAGEN).

    Techniques: Isolation, SDS Page, Silver Staining, Western Blot, Infection, Quantitative RT-PCR, Transfection, In Vitro, Incubation, Immunoprecipitation, Expressing

    Regulatory Mechanism of lncITPF in Its Host Gene (A) Itgbl1 expression was higher in fibrotic tissue than in normal tissue on the basis of RNA sequencing. (B) Itgbl1 was upregulated in cells treated with 5 ng/mL TGF-β1 for 12, 24, 48, and 72 hr. (C) qRT-PCR analysis showed that Itgbl1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (D) Western blot showed that ITGBL1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (E) Single-molecule RNA-FISH detecting the location of lncITPF (red) in cells. U6 and 18S RNA were used as cytoplasmic and nuclear localization markers, respectively. DNA (blue) was stained with DAPI. A representative image is shown. (F) qRT-PCR analysis of RNAs purified from nucleoplasmic (red), chromatin nuclear (gray), and cytosolic (blue) compartments in cells. NC indicates a negative control, BP indicates blank plasmid, and RP indicates the recombinant plasmid of the overexpressed lncITPF. Each bar represents the mean ± SD; n = 6; **p

    Journal: Molecular Therapy

    Article Title: lncITPF Promotes Pulmonary Fibrosis by Targeting hnRNP-L Depending on Its Host Gene ITGBL1

    doi: 10.1016/j.ymthe.2018.08.026

    Figure Lengend Snippet: Regulatory Mechanism of lncITPF in Its Host Gene (A) Itgbl1 expression was higher in fibrotic tissue than in normal tissue on the basis of RNA sequencing. (B) Itgbl1 was upregulated in cells treated with 5 ng/mL TGF-β1 for 12, 24, 48, and 72 hr. (C) qRT-PCR analysis showed that Itgbl1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (D) Western blot showed that ITGBL1 was decreased by lncITPF RNAi and increased by lncITPF overexpression. (E) Single-molecule RNA-FISH detecting the location of lncITPF (red) in cells. U6 and 18S RNA were used as cytoplasmic and nuclear localization markers, respectively. DNA (blue) was stained with DAPI. A representative image is shown. (F) qRT-PCR analysis of RNAs purified from nucleoplasmic (red), chromatin nuclear (gray), and cytosolic (blue) compartments in cells. NC indicates a negative control, BP indicates blank plasmid, and RP indicates the recombinant plasmid of the overexpressed lncITPF. Each bar represents the mean ± SD; n = 6; **p

    Article Snippet: lncITPF transcripts were transcribed using T7 and SP6 RNA polymerase (Ambion) in vitro , then by using the RNeasy Plus Mini Kit (QIAGEN, Germany) and treated with TURBO DNase I. Purified RNAs were biotin-labeled using Biotin RNA Labeling Mix (Ambion Life).

    Techniques: Expressing, RNA Sequencing Assay, Quantitative RT-PCR, Over Expression, Western Blot, Fluorescence In Situ Hybridization, Staining, Purification, Negative Control, Plasmid Preparation, Recombinant

    Downstream Mechanism of lncITPF (A) SDS-PAGE analysis of proteins purified from in vitro binding assay using biotinylated lncITPF or antisense control RNA. The arrow indicates that the highlighted protein bands were subjected to LC-MS. (B) Protein levels of hnRNP-L in immunoprecipitates with lncITPF RNA were evaluated by western blot. (C) RNA immunoprecipitation assays followed by qRT-PCR analysis of co-purified RNAs with hnRNP-L in cross-linked cells. lncITPF RNA expression levels are presented as fold enrichment values relative to IgG immunoprecipitates. (D) Overexpression of lncITPF increased Itgbl1 mRNA expression, which was impaired in hnRNP-L knockdown. (E) Overexpression of lncITPF increased ITGBL1 protein expression, which was impaired in hnRNP-L knockdown. (F) ChIP-qPCR analysis of hnRNP-L occupancy in ITGBL1 promoter in cells. (G) The ChIP-qPCR gel images were quantified and analyzed. (H) ChIP analysis of the effect of hnRNP-L on H3 and H4 histone acetylation of ITGBL1 promoter regions. NC indicates a negative control, BP indicates blank plasmid, and RP indicates the recombinant plasmid of the overexpressed lncITPF. Each bar represents the mean ± SD; n = 6; *p

    Journal: Molecular Therapy

    Article Title: lncITPF Promotes Pulmonary Fibrosis by Targeting hnRNP-L Depending on Its Host Gene ITGBL1

    doi: 10.1016/j.ymthe.2018.08.026

    Figure Lengend Snippet: Downstream Mechanism of lncITPF (A) SDS-PAGE analysis of proteins purified from in vitro binding assay using biotinylated lncITPF or antisense control RNA. The arrow indicates that the highlighted protein bands were subjected to LC-MS. (B) Protein levels of hnRNP-L in immunoprecipitates with lncITPF RNA were evaluated by western blot. (C) RNA immunoprecipitation assays followed by qRT-PCR analysis of co-purified RNAs with hnRNP-L in cross-linked cells. lncITPF RNA expression levels are presented as fold enrichment values relative to IgG immunoprecipitates. (D) Overexpression of lncITPF increased Itgbl1 mRNA expression, which was impaired in hnRNP-L knockdown. (E) Overexpression of lncITPF increased ITGBL1 protein expression, which was impaired in hnRNP-L knockdown. (F) ChIP-qPCR analysis of hnRNP-L occupancy in ITGBL1 promoter in cells. (G) The ChIP-qPCR gel images were quantified and analyzed. (H) ChIP analysis of the effect of hnRNP-L on H3 and H4 histone acetylation of ITGBL1 promoter regions. NC indicates a negative control, BP indicates blank plasmid, and RP indicates the recombinant plasmid of the overexpressed lncITPF. Each bar represents the mean ± SD; n = 6; *p

    Article Snippet: lncITPF transcripts were transcribed using T7 and SP6 RNA polymerase (Ambion) in vitro , then by using the RNeasy Plus Mini Kit (QIAGEN, Germany) and treated with TURBO DNase I. Purified RNAs were biotin-labeled using Biotin RNA Labeling Mix (Ambion Life).

    Techniques: SDS Page, Purification, In Vitro, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Immunoprecipitation, Quantitative RT-PCR, RNA Expression, Over Expression, Expressing, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control, Plasmid Preparation, Recombinant

    ELK4 expression in Aβ 1-42 -incubated ECs and ELK4 regulated BBB permeability in AD microenvironment. (A) RNA microarray analysis was performed in ECs treated with shLINC00662. Red indicates high relative expression and green indicates low relative expression. (B) Relative expression levels of ZEB1, ELK4, and ART1 determined by qRT-PCR. Data represent mean ± SD (n = 3, each). **P

    Journal: bioRxiv

    Article Title: TRA2A-induced upregulation of LINC00662 regulates blood-brain barrier permeability by affecting ELK4 mRNA stability in Alzheimer’s microenvironment

    doi: 10.1101/848408

    Figure Lengend Snippet: ELK4 expression in Aβ 1-42 -incubated ECs and ELK4 regulated BBB permeability in AD microenvironment. (A) RNA microarray analysis was performed in ECs treated with shLINC00662. Red indicates high relative expression and green indicates low relative expression. (B) Relative expression levels of ZEB1, ELK4, and ART1 determined by qRT-PCR. Data represent mean ± SD (n = 3, each). **P

    Article Snippet: RNA pull-down assays Biotin-labelled, full length LINC00662, or antisense RNA was prepared with the Biotin RNA Labeling Mix (GenePharma, Shanghai, China) and transfected into ECs.

    Techniques: Expressing, Incubation, Permeability, Microarray, Quantitative RT-PCR

    Atrolnc‐1 regulates MuRF‐1 via interacting with ABIN‐1 in vitro and in vivo . (A) In situ hybridization analysis with a biotin‐labelled Atrolnc‐1 antisense probe in C2C12 cell. Atrolnc‐1 positive signals manifest as brown granules located both in the nuclear (N) and cytoplasm (Cyto). Atrolnc‐1 sense probe was used as negative control (CTL). Scale bar = 20 μm. (B) SDS‐PAGE gels exhibit the results of RNA pull‐down assay. The interaction of biotin‐labelled Atrolnc‐1 probe (marked as 2) with proteins from cytoplasm (left gel) and nucleus (right gel) extracted from C2C12 cells are shown. Atrolnc‐1 antisense probe was used as control (marked as 1). M represented molecular marker. (C) The table lists mass spectrometry result of proteins interacting with Atrolnc‐1. In the gels in (B), ABIN‐1, an inhibitor of NF‐kB signalling, was found to bind with biotin‐labelled Atrolnc‐1 probe in cytoplasmic extracts from myotubes. The iBAQ denotes the sum of all the peptides intensities divided by the number of observable peptides of a protein. (D) Left panel: a representative immunoblotting shows a band at 72 kD using ABIN‐1 antibody in the proteins pulled‐down by biotin‐labelled Atrolnc‐1 probe. Right panel: in cytoplasmic extracts of tibialis anterior (TA) muscle from mice, immunoblotting also detected a band at 72 kD in RNA pull‐down assay by biotin‐labelled Atrolnc‐1 probe. (E) The result of immunoprecipitation (IP) in TA muscle from mice with or without chronic kidney disease (CKD). Ten per cent of the IP product was used to conduct western blot (low panel), showing a band at 72 kD using ABIN‐1 antibody (ABIN‐1 Ab) while a RT‐PCR of Arolnc‐1(upper panel) was performed using the rest of IP product. Notice that the Atrolnc‐1 interacted with ABIN‐1 was increased in CKD mice. Mock, normal mouse IgG. (F) Immunoblot in upper panel shows increased ABIN‐1 expression in C2C12 myotubes after transduced by CRISPR/dCas9 mediated synergistic activation mediator transcription activation system. Bar graph (low panel) illustrates the quantity result (mean ± SEM; n = 5, * P

    Journal: Journal of Cachexia, Sarcopenia and Muscle

    Article Title: Long‐noncoding RNA Atrolnc‐1 promotes muscle wasting in mice with chronic kidney disease) Long‐noncoding RNA Atrolnc‐1 promotes muscle wasting in mice with chronic kidney disease

    doi: 10.1002/jcsm.12321

    Figure Lengend Snippet: Atrolnc‐1 regulates MuRF‐1 via interacting with ABIN‐1 in vitro and in vivo . (A) In situ hybridization analysis with a biotin‐labelled Atrolnc‐1 antisense probe in C2C12 cell. Atrolnc‐1 positive signals manifest as brown granules located both in the nuclear (N) and cytoplasm (Cyto). Atrolnc‐1 sense probe was used as negative control (CTL). Scale bar = 20 μm. (B) SDS‐PAGE gels exhibit the results of RNA pull‐down assay. The interaction of biotin‐labelled Atrolnc‐1 probe (marked as 2) with proteins from cytoplasm (left gel) and nucleus (right gel) extracted from C2C12 cells are shown. Atrolnc‐1 antisense probe was used as control (marked as 1). M represented molecular marker. (C) The table lists mass spectrometry result of proteins interacting with Atrolnc‐1. In the gels in (B), ABIN‐1, an inhibitor of NF‐kB signalling, was found to bind with biotin‐labelled Atrolnc‐1 probe in cytoplasmic extracts from myotubes. The iBAQ denotes the sum of all the peptides intensities divided by the number of observable peptides of a protein. (D) Left panel: a representative immunoblotting shows a band at 72 kD using ABIN‐1 antibody in the proteins pulled‐down by biotin‐labelled Atrolnc‐1 probe. Right panel: in cytoplasmic extracts of tibialis anterior (TA) muscle from mice, immunoblotting also detected a band at 72 kD in RNA pull‐down assay by biotin‐labelled Atrolnc‐1 probe. (E) The result of immunoprecipitation (IP) in TA muscle from mice with or without chronic kidney disease (CKD). Ten per cent of the IP product was used to conduct western blot (low panel), showing a band at 72 kD using ABIN‐1 antibody (ABIN‐1 Ab) while a RT‐PCR of Arolnc‐1(upper panel) was performed using the rest of IP product. Notice that the Atrolnc‐1 interacted with ABIN‐1 was increased in CKD mice. Mock, normal mouse IgG. (F) Immunoblot in upper panel shows increased ABIN‐1 expression in C2C12 myotubes after transduced by CRISPR/dCas9 mediated synergistic activation mediator transcription activation system. Bar graph (low panel) illustrates the quantity result (mean ± SEM; n = 5, * P

    Article Snippet: To obtain the biotin‐labelled Atrolnc‐1‐RNA probe, linearized pBlue‐Atrolnc‐1 plasmid was incubated with biotin RNA labelling mix (Roche Molecular Systems, Inc. Hague Road, IN) for 20 min at 37°C, in vitro transcription was performed using a MAXIscript® T7/T3 Transcription Kit (Thermo Fisher, Waltham, MA, USA) following manufacturer's instructions.

    Techniques: In Vitro, In Vivo, In Situ Hybridization, Negative Control, CTL Assay, SDS Page, Pull Down Assay, Marker, Mass Spectrometry, Mouse Assay, Immunoprecipitation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, CRISPR, Activation Assay

    In situ hybridization of rice root apices with probes specific for transcripts of cdc2Os1 , cdc2Os2 , cdc2Os3 , and R2 . Longitudinal sections of roots were allowed to hybridize with digoxigenin-labeled RNA probes. Hybridization signals are visible as brownish-purple staining. a, cdc2Os1 antisense probe; b, cdc2Os2 antisense probe; c, cdc2Os3 antisense probe; d, R2 antisense probe; and e, cdc2Os1 sense probe. Bar = 100 μm.

    Journal: Plant Physiology

    Article Title: Differential Expression of Genes for Cyclin-Dependent Protein Kinases in Rice Plants 1

    doi:

    Figure Lengend Snippet: In situ hybridization of rice root apices with probes specific for transcripts of cdc2Os1 , cdc2Os2 , cdc2Os3 , and R2 . Longitudinal sections of roots were allowed to hybridize with digoxigenin-labeled RNA probes. Hybridization signals are visible as brownish-purple staining. a, cdc2Os1 antisense probe; b, cdc2Os2 antisense probe; c, cdc2Os3 antisense probe; d, R2 antisense probe; and e, cdc2Os1 sense probe. Bar = 100 μm.

    Article Snippet: Digoxigenin- and biotin-labeled probes were generated with a digoxigenin RNA-labeling kit in combination with a digoxigenin RNA-labeling mix and a biotin RNA-labeling mix, respectively (Boehringer Mannheim).

    Techniques: In Situ Hybridization, Labeling, Hybridization, Staining

    EPIC1 is a nuclear lncRNA regulating MYC targets expression (A) qRT-PCR analysis of EPIC1 expression (top) and Western blot (bottom) of subcellular fractionation in MCF-7cells. GAPDH and U6 RNA served as a marker for cytoplasmic and nuclear gene localization, respectively. SNRP70 and GAPDH served as a specific nuclear and cytoplasmic marker to whole cell lysates (WCL), cytoplasmic (Cyto), and nuclear fractionation (Nuc). Error bars indicate mean ± SD, n = 3 for technical replicates. (B) Schematic of the identification of EPIC1 correlated genes in breast tumors from TCGA (yellow), and genes potentially regulated by EPIC1 in MCF-7 cells (green). (C) Co-expression analysis showing that EPIC1 expression is associated with 2005 genes in 559 patients with breast cancer (BRCA). Each column represents one patient. (D) GSEA analysis of the EPIC1 -related pathways in 20 cancer types (left panel) and EPIC1 knockdown MCF-7 cells (right panel). The heatmap indicates the GSEA scores. (E) Association between the enrichment of MYC targets and EPIC1 expression in breast tumors by GSEA analysis (D). (F) EPIC1 -regulated gene expression by qRT-PCR analysis (top) and RNA-seq (bottom). Error bars indicate mean ± SD, n = 3 for technical replicates. (G) Western blot of MYC-regulated targets in MCF-7 (left) and ZR-75-1 (right) cells treated with EPIC1 and MYC siRNAs. .

    Journal: Cancer cell

    Article Title: LncRNA epigenetic landscape analysis identifies EPIC1 as an oncogenic lncRNA that interacts with MYC and promotes cell cycle progression in cancer

    doi: 10.1016/j.ccell.2018.03.006

    Figure Lengend Snippet: EPIC1 is a nuclear lncRNA regulating MYC targets expression (A) qRT-PCR analysis of EPIC1 expression (top) and Western blot (bottom) of subcellular fractionation in MCF-7cells. GAPDH and U6 RNA served as a marker for cytoplasmic and nuclear gene localization, respectively. SNRP70 and GAPDH served as a specific nuclear and cytoplasmic marker to whole cell lysates (WCL), cytoplasmic (Cyto), and nuclear fractionation (Nuc). Error bars indicate mean ± SD, n = 3 for technical replicates. (B) Schematic of the identification of EPIC1 correlated genes in breast tumors from TCGA (yellow), and genes potentially regulated by EPIC1 in MCF-7 cells (green). (C) Co-expression analysis showing that EPIC1 expression is associated with 2005 genes in 559 patients with breast cancer (BRCA). Each column represents one patient. (D) GSEA analysis of the EPIC1 -related pathways in 20 cancer types (left panel) and EPIC1 knockdown MCF-7 cells (right panel). The heatmap indicates the GSEA scores. (E) Association between the enrichment of MYC targets and EPIC1 expression in breast tumors by GSEA analysis (D). (F) EPIC1 -regulated gene expression by qRT-PCR analysis (top) and RNA-seq (bottom). Error bars indicate mean ± SD, n = 3 for technical replicates. (G) Western blot of MYC-regulated targets in MCF-7 (left) and ZR-75-1 (right) cells treated with EPIC1 and MYC siRNAs. .

    Article Snippet: Biotin-labeled full-length and truncated fragments of EPIC1 RNA were transcribed in vitro with a Biotin RNA Labeling Mix Kit (Roche, #11685597910) and T7 RNA polymerase (Roche, #10881775001) using the PCR products as a template, treated with RNase-free DNase I (Promega, #M198A), and then isolated with an RNeasy Mini kit.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Fractionation, Marker, RNA Sequencing Assay

    EGFR-AS1 promotes the maintenance of EGFR mRNA stability by binding to HuR. a Silver staining SDS-PAGE gel of electrophoretically separated proteins immunoprecipitated with EGFR-AS1 and its antisense RNA in 786-O cells. b RNA pull-down assay was performed in 786-O and A498 cells using biotinylated EGFR-AS1 or antisense RNA probe transcribed in vitro and detected by western blots. c Upper: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR-AS1 RNA enrichment in immunoprecipitated complexes. IgG is the negative control. Lower: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR RNA enrichment in immunoprecipitated complexes. d RIP assay of the enrichment of EGFR mRNA with HuR between the EGFR-AS1 knockdown and NC group in RCC cells. IgG was used as an internal control. e Upper: RNA FISH analysis of EGFR mRNA (green) and immunofluorescence detection of HuR (red) in RCC cells. The rightmost graph shows colocalization between the green signal (EGFR) and the red signal (HuR). Pearson’s R = 0.583. Scale bar = 50 μm. Lower: RNA FISH analysis of EGFR-AS1 (green) and immunofluorescence detection of HuR (red) in RCC cells. Pearson’s R = 0.416. f The rate of degradation of the EGFR mRNA between the HuR knockdown and control group using RNA stability assays in RCC cells. g The rate of degradation of the EGFR mRNA between the HuR overexpressing and control group using RNA stability assays in RCC cells. h The rate of degradation of the EGFR mRNA in the EGFR-AS1 knockdown and control cells transfected with pcDNA3.1 + -HuR over 12 h in KETR-3 and ACHN cells. i The rate of degradation of the EGFR mRNA in the EGFR-AS1 overexpressing and control cells transfected with HuR siRNA over 12 h

    Journal: Cell Death & Disease

    Article Title: Long noncoding RNA EGFR-AS1 promotes cell growth and metastasis via affecting HuR mediated mRNA stability of EGFR in renal cancer

    doi: 10.1038/s41419-019-1331-9

    Figure Lengend Snippet: EGFR-AS1 promotes the maintenance of EGFR mRNA stability by binding to HuR. a Silver staining SDS-PAGE gel of electrophoretically separated proteins immunoprecipitated with EGFR-AS1 and its antisense RNA in 786-O cells. b RNA pull-down assay was performed in 786-O and A498 cells using biotinylated EGFR-AS1 or antisense RNA probe transcribed in vitro and detected by western blots. c Upper: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR-AS1 RNA enrichment in immunoprecipitated complexes. IgG is the negative control. Lower: RIP assays were performed in 786-O cells using HuR antibody to detect EGFR RNA enrichment in immunoprecipitated complexes. d RIP assay of the enrichment of EGFR mRNA with HuR between the EGFR-AS1 knockdown and NC group in RCC cells. IgG was used as an internal control. e Upper: RNA FISH analysis of EGFR mRNA (green) and immunofluorescence detection of HuR (red) in RCC cells. The rightmost graph shows colocalization between the green signal (EGFR) and the red signal (HuR). Pearson’s R = 0.583. Scale bar = 50 μm. Lower: RNA FISH analysis of EGFR-AS1 (green) and immunofluorescence detection of HuR (red) in RCC cells. Pearson’s R = 0.416. f The rate of degradation of the EGFR mRNA between the HuR knockdown and control group using RNA stability assays in RCC cells. g The rate of degradation of the EGFR mRNA between the HuR overexpressing and control group using RNA stability assays in RCC cells. h The rate of degradation of the EGFR mRNA in the EGFR-AS1 knockdown and control cells transfected with pcDNA3.1 + -HuR over 12 h in KETR-3 and ACHN cells. i The rate of degradation of the EGFR mRNA in the EGFR-AS1 overexpressing and control cells transfected with HuR siRNA over 12 h

    Article Snippet: RNA pull-down assay and mass spectrometry LncRNA EGFR-AS1 was transcribed in vitro from the vector pSPT19-lncRNA-EGFR-AS1 and biotinylated with biotinylated RNA labeling mix (Roche, Mannheim, Germany) and T7 RNA polymerase (Roche) and treated with RNase-free DNase I (Roche).

    Techniques: Binding Assay, Silver Staining, SDS Page, Immunoprecipitation, Pull Down Assay, In Vitro, Western Blot, Negative Control, Fluorescence In Situ Hybridization, Immunofluorescence, Transfection

    Number of detected probes and probe sets in hybridizations using RNA from four tumor samples labeled with the FlashTag Biotin- and FlashTag Biotin HSR labeling kits .

    Journal: BMC Research Notes

    Article Title: Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC)

    doi: 10.1186/1756-0500-4-424

    Figure Lengend Snippet: Number of detected probes and probe sets in hybridizations using RNA from four tumor samples labeled with the FlashTag Biotin- and FlashTag Biotin HSR labeling kits .

    Article Snippet: Different chip lot numbers Correlations in signal intensities were examined across two different lots of arrays that were hybridized (in triplicates) to 100 ng of labeled RNA (FlashTag™ Biotin RNA Labeling Kit, ATP-mix dilution 1:50) from of a single RNA preparation of a T2 NSCLC tumor using the HP Kit .

    Techniques: Labeling

    PCA for a 12 gene signature for prognosis in NSCLC in hybridizations using RNA from four tumor samples labeled with both the FlashTag Biotin- and FlashTag Biotin HSR labeling kits . F (red) represents samples labeled with the FlashTag Biotin kit, and, FH (green) represents the same samples labeled with the FlashTag Biotin HSR labeling kit.

    Journal: BMC Research Notes

    Article Title: Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC)

    doi: 10.1186/1756-0500-4-424

    Figure Lengend Snippet: PCA for a 12 gene signature for prognosis in NSCLC in hybridizations using RNA from four tumor samples labeled with both the FlashTag Biotin- and FlashTag Biotin HSR labeling kits . F (red) represents samples labeled with the FlashTag Biotin kit, and, FH (green) represents the same samples labeled with the FlashTag Biotin HSR labeling kit.

    Article Snippet: Different chip lot numbers Correlations in signal intensities were examined across two different lots of arrays that were hybridized (in triplicates) to 100 ng of labeled RNA (FlashTag™ Biotin RNA Labeling Kit, ATP-mix dilution 1:50) from of a single RNA preparation of a T2 NSCLC tumor using the HP Kit .

    Techniques: Labeling

    Mean signal and background intensities in hybridizations using RNA from four tumor samples labeled with the FlashTag Biotin- and FlashTag Biotin HSR labeling kits .

    Journal: BMC Research Notes

    Article Title: Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC)

    doi: 10.1186/1756-0500-4-424

    Figure Lengend Snippet: Mean signal and background intensities in hybridizations using RNA from four tumor samples labeled with the FlashTag Biotin- and FlashTag Biotin HSR labeling kits .

    Article Snippet: Different chip lot numbers Correlations in signal intensities were examined across two different lots of arrays that were hybridized (in triplicates) to 100 ng of labeled RNA (FlashTag™ Biotin RNA Labeling Kit, ATP-mix dilution 1:50) from of a single RNA preparation of a T2 NSCLC tumor using the HP Kit .

    Techniques: Labeling

    PCA for all human non-coding RNAs in hybridizations using RNA from four tumor samples labeled with both the FlashTag Biotin- and FlashTag Biotin HSR labeling kits . F (red) represents samples labeled with the FlashTag Biotin kit, and, FH (green) represents the same samples labeled with the FlashTag Biotin HSR labeling kit.

    Journal: BMC Research Notes

    Article Title: Analytical variables influencing the performance of a miRNA based laboratory assay for prediction of relapse in stage I non-small cell lung cancer (NSCLC)

    doi: 10.1186/1756-0500-4-424

    Figure Lengend Snippet: PCA for all human non-coding RNAs in hybridizations using RNA from four tumor samples labeled with both the FlashTag Biotin- and FlashTag Biotin HSR labeling kits . F (red) represents samples labeled with the FlashTag Biotin kit, and, FH (green) represents the same samples labeled with the FlashTag Biotin HSR labeling kit.

    Article Snippet: Different chip lot numbers Correlations in signal intensities were examined across two different lots of arrays that were hybridized (in triplicates) to 100 ng of labeled RNA (FlashTag™ Biotin RNA Labeling Kit, ATP-mix dilution 1:50) from of a single RNA preparation of a T2 NSCLC tumor using the HP Kit .

    Techniques: Labeling