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  • 99
    TaKaRa pmcherry c1
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 24832 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc pmcherry c1
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pmcherry c1/product/Addgene inc
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    93
    Becton Dickinson pmcherry c1
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa pmcherry c1 template
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1 Template, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Addgene inc pmcherry c1 vector
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1 Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Addgene inc pmcherry c1 plasmids
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1 Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Addgene inc pmcherry c1 fak ha
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1 Fak Ha, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson pmcherry c1 vector
    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with <t>pmCherry-AUF1</t> (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), <t>pmCherry-C1</t> (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P
    Pmcherry C1 Vector, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with pmCherry-AUF1 (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), pmCherry-C1 (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P

    Journal: Oncotarget

    Article Title: RNA-binding protein AUF1 suppresses miR-122 biogenesis by down-regulating Dicer1 in hepatocellular carcinoma

    doi: 10.18632/oncotarget.24079

    Figure Lengend Snippet: Inhibited expression of AUF1 promotes HCC cell death ( A ) PLC/PRF/5 cells were grown in 6-well plate. The cells were transfected with pmCherry-AUF1 (AUF1) or siRNA of AUF1 (siAUF1) for 24 h in the medium without serum. Apoptosis was determined by flow cytometry. Control cells were transfected with liposome (NC), pmCherry-C1 (C1) or siMock. ( B ) 80% confluent HHL-5 cells were transfected with siAUF1 or siMock for 24 h in the medium without serum. The expression of Dicer1, AUF1, and cleaved PARP1 was determined by Western blotting (a). The relative expression levels are presented as mean ± SD (b, c, d). n = 4, * P

    Article Snippet: The constructed pEGFP-AUF1 was verified by sequencing. pmCherry-AUF1 was constructed by inserting the sequence of p45AUF1 into pmCherry-C1 (Clontech, Mountain View, CA).

    Techniques: Expressing, Planar Chromatography, Transfection, Flow Cytometry, Cytometry, Western Blot

    Subcellular distribution of cloned TRIM5 group proteins. ( A ) Domain composition of TRIM5 group proteins. The protein accession numbers in GenBank are indicated. ( B ) 293T cells (1 × 10 6 ) were transfected with the plasmids containing V5-tagged TRIM cDNAs (2 μg) for 30 h, and whole-cell extracts (WCE, 50 μg) were resolved on on 4–12% NuPAGE ( upper panel ) or 10% SDS-PAGE ( lower panel ) for immunoblotting assay with anti-V5 Ab. ( C ) NIH3T3 cells were transfected with indicated mCherry florescence protein–tagged TRIM genes cloned in pmCherry-C1 vector (1 μg) for 24 h and visualized by fluorescence microscopy at ×400 magnification. The representative images for each TRIM protein are shown. Of > 200 cells inspected, > 90% of TRIM12c-transfected cells exhibited cytoplasmic body–like structures.

    Journal: The Journal of Immunology Author Choice

    Article Title: Tripartite Motif (TRIM) 12c, a Mouse Homolog of TRIM5, Is a Ubiquitin Ligase That Stimulates Type I IFN and NF-κB Pathways along with TNFR-Associated Factor 6

    doi: 10.4049/jimmunol.1402064

    Figure Lengend Snippet: Subcellular distribution of cloned TRIM5 group proteins. ( A ) Domain composition of TRIM5 group proteins. The protein accession numbers in GenBank are indicated. ( B ) 293T cells (1 × 10 6 ) were transfected with the plasmids containing V5-tagged TRIM cDNAs (2 μg) for 30 h, and whole-cell extracts (WCE, 50 μg) were resolved on on 4–12% NuPAGE ( upper panel ) or 10% SDS-PAGE ( lower panel ) for immunoblotting assay with anti-V5 Ab. ( C ) NIH3T3 cells were transfected with indicated mCherry florescence protein–tagged TRIM genes cloned in pmCherry-C1 vector (1 μg) for 24 h and visualized by fluorescence microscopy at ×400 magnification. The representative images for each TRIM protein are shown. Of > 200 cells inspected, > 90% of TRIM12c-transfected cells exhibited cytoplasmic body–like structures.

    Article Snippet: Full-length cDNAs for Trim5 group genes were cloned from RAW 264.7 cells or BMDCs (both 1 × 106 cells) stimulated with IFN-γ (100 U/ml) for 24 h. First-strand cDNAs were synthesized from 5 μg total RNA prepared by the RNeasy kit (Qiagen) or TRIzol (Invitrogen) and used to amplify full-length cDNA for Trim genes by conventional PCR and cloned into the pcDNA3.1-V5 vector or pmCherry-C1 vector (Clontech) using primers listed in . pcDNA3.1 vectors (Invitrogen) containing Flag-tagged mouse IRF3 or IRF7 were described ( , ).

    Techniques: Clone Assay, Transfection, SDS Page, Plasmid Preparation, Fluorescence, Microscopy

    KANK1-talin interaction is required for recruiting CMSCs to FAs. Widefield fluorescence images of HeLa cells depleted of KANK1 and KANK2 and expressing the indicated siRNA-resistant GFP-KANK1 fusions (rescue), stained for the FA marker phospho-tyrosine (pY). Widefield fluorescence images of HeLa cells transfected with the control siRNA or siRNAs against KANK1 and KANK2, expressing GFP alone or the indicated siRNA-resistant GFP-KANK1 fusions and stained for LL5β or KIF21A. Quantification of peripheral clustering of LL5β and KIF21A in cells treated as in panel (B) (n=12, 5–6 cells per condition). TIRFM images of live HeLa cells depleted of KANK1 and KANK2 and co-expressing the indicated siRNA-resistant GFP-KANK1 fusions and mCherry-CLASP2. Quantification of peripheral clustering of mCherry-CLASP2 in cells treated as in panel (D) (n=20, 8 cells per condition). Widefield fluorescence images of HeLa cells transfected with the indicated GFP-KANK1 fusions and stained for endogenous LL5β. Quantification of peripheral clustering of LL5β in cells treated as in panel (F) (n=12, 6 cells per condition). Widefield fluorescence images of HeLa cells transfected with GFP-tagged KANK1 or its CC1 mutant and stained for LL5β. Quantification of peripheral clustering of LL5β cells treated as in panel (H) (n=12, 6 cells per condition). Error bars, SEM; ns, non-significant; **, P

    Journal: bioRxiv

    Article Title: Talin-KANK1 interaction controls the recruitment of cortical microtubule stabilizing complexes to focal adhesions

    doi: 10.1101/055210

    Figure Lengend Snippet: KANK1-talin interaction is required for recruiting CMSCs to FAs. Widefield fluorescence images of HeLa cells depleted of KANK1 and KANK2 and expressing the indicated siRNA-resistant GFP-KANK1 fusions (rescue), stained for the FA marker phospho-tyrosine (pY). Widefield fluorescence images of HeLa cells transfected with the control siRNA or siRNAs against KANK1 and KANK2, expressing GFP alone or the indicated siRNA-resistant GFP-KANK1 fusions and stained for LL5β or KIF21A. Quantification of peripheral clustering of LL5β and KIF21A in cells treated as in panel (B) (n=12, 5–6 cells per condition). TIRFM images of live HeLa cells depleted of KANK1 and KANK2 and co-expressing the indicated siRNA-resistant GFP-KANK1 fusions and mCherry-CLASP2. Quantification of peripheral clustering of mCherry-CLASP2 in cells treated as in panel (D) (n=20, 8 cells per condition). Widefield fluorescence images of HeLa cells transfected with the indicated GFP-KANK1 fusions and stained for endogenous LL5β. Quantification of peripheral clustering of LL5β in cells treated as in panel (F) (n=12, 6 cells per condition). Widefield fluorescence images of HeLa cells transfected with GFP-tagged KANK1 or its CC1 mutant and stained for LL5β. Quantification of peripheral clustering of LL5β cells treated as in panel (H) (n=12, 6 cells per condition). Error bars, SEM; ns, non-significant; **, P

    Article Snippet: Rescue constructs for BioGFP-tagged KANK1 were either based on the version previously described ( ) or a version obtained by PCR-based mutagenesis of the sequence AGTCAGCGTCTGCGAA to GGTGAGTGTGTGTGAG. mCherry-tagged paxillin construct was made by replacing GFP from pQC-GPXN ( ) by mCherry (pmCherry-C1, Clontech).

    Techniques: Fluorescence, Expressing, Staining, Marker, Transfection, Mutagenesis

    The KN motif of KANK1 interacts with the R7 domain of talin1. Schematic representation of KANK1 and the deletion mutants used in this study, and the summary of their interactions and localization. N.d., not determined in this study. TIRFM images of live HeLa cells transiently expressing the indicated GFP-tagged KANK1 deletion mutants together with the focal adhesion marker mCherry-paxillin. Identification of the binding partners of Bio-GFP-tagged KANK1 and its indicated deletion mutants by using streptavidin pull down assays from HEK293T cells combined with mass spectrometry. Streptavidin pull down assays with the BioGFP-tagged KANK1 or the indicated KANK1 mutants, co-expressed with GFP-talin1 in HEK293T cells, analyzed by Western blotting with the indicated antibodies. Sequence alignment of KANK1 and KANK2 with the known talin-binding sites of DLC1, RIAM and Paxillin. The LD-motif and the interacting hydrophobic residues are highlighted green and blue respectively. Schematic representation of talin1 and the deletion mutants used in this study, and their interaction with KANK1. Streptavidin pull down assays with the BioGFP-tagged talin1 or the indicated talin1 mutants, co-expressed with HA-KANK1 in HEK293T cells, analyzed by Western blotting with the indicated antibodies.

    Journal: bioRxiv

    Article Title: Talin-KANK1 interaction controls the recruitment of cortical microtubule stabilizing complexes to focal adhesions

    doi: 10.1101/055210

    Figure Lengend Snippet: The KN motif of KANK1 interacts with the R7 domain of talin1. Schematic representation of KANK1 and the deletion mutants used in this study, and the summary of their interactions and localization. N.d., not determined in this study. TIRFM images of live HeLa cells transiently expressing the indicated GFP-tagged KANK1 deletion mutants together with the focal adhesion marker mCherry-paxillin. Identification of the binding partners of Bio-GFP-tagged KANK1 and its indicated deletion mutants by using streptavidin pull down assays from HEK293T cells combined with mass spectrometry. Streptavidin pull down assays with the BioGFP-tagged KANK1 or the indicated KANK1 mutants, co-expressed with GFP-talin1 in HEK293T cells, analyzed by Western blotting with the indicated antibodies. Sequence alignment of KANK1 and KANK2 with the known talin-binding sites of DLC1, RIAM and Paxillin. The LD-motif and the interacting hydrophobic residues are highlighted green and blue respectively. Schematic representation of talin1 and the deletion mutants used in this study, and their interaction with KANK1. Streptavidin pull down assays with the BioGFP-tagged talin1 or the indicated talin1 mutants, co-expressed with HA-KANK1 in HEK293T cells, analyzed by Western blotting with the indicated antibodies.

    Article Snippet: Rescue constructs for BioGFP-tagged KANK1 were either based on the version previously described ( ) or a version obtained by PCR-based mutagenesis of the sequence AGTCAGCGTCTGCGAA to GGTGAGTGTGTGTGAG. mCherry-tagged paxillin construct was made by replacing GFP from pQC-GPXN ( ) by mCherry (pmCherry-C1, Clontech).

    Techniques: Expressing, Marker, Binding Assay, Mass Spectrometry, Western Blot, Sequencing