pegfp c1 Takara Search Results


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  • 99
    TaKaRa pegfp c1
    Postmortem histology of tumors after intravenous delivery of either pCMV-TK (a, c, d) or <t>pEGFP-C1</t> (b)-loaded microbubbles and treatment with ultrasound and GCV. (a, b) Acellular zones ( arrows ) visible under H E stain. (c) Merged image of two sister
    Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 898 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    egfp  (TaKaRa)
    97
    TaKaRa egfp
    Postmortem histology of tumors after intravenous delivery of either pCMV-TK (a, c, d) or <t>pEGFP-C1</t> (b)-loaded microbubbles and treatment with ultrasound and GCV. (a, b) Acellular zones ( arrows ) visible under H E stain. (c) Merged image of two sister
    Egfp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 97/100, based on 7452 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    TaKaRa bglii hindiii digested pegfp c1
    Postmortem histology of tumors after intravenous delivery of either pCMV-TK (a, c, d) or <t>pEGFP-C1</t> (b)-loaded microbubbles and treatment with ultrasound and GCV. (a, b) Acellular zones ( arrows ) visible under H E stain. (c) Merged image of two sister
    Bglii Hindiii Digested Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa plasmids pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Plasmids Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 237 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa bglii ecori digested pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Bglii Ecori Digested Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    TaKaRa cdnas pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Cdnas Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa complementary dnas pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Complementary Dnas Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa sali digested pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Sali Digested Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 83/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    TaKaRa expression plasmid pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Expression Plasmid Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa plasmid dna pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Plasmid Dna Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    TaKaRa methuosis pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Methuosis Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa clontech pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Clontech Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa mammalian expression vector pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Mammalian Expression Vector Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa site directed mutagenesis plasmid pegfp c1
    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid <t>pEGFP</t> C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.
    Site Directed Mutagenesis Plasmid Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    TaKaRa mammalian expression vectors pegfp c1
    Cellular localizations of low-risk and high-risk HPV E6 in COS-1 cells. COS-1 cells were transfected with plasmids pZMZ66 (6E6E7, nt 85 to 939), pZMZ67 (11E6E7, nt 85 to 924), pZMZ69 (18E6E7, nt 103 to 967), pZMZ70 (16E6E7, nt 81 to 880), and pZMZ81 (16E6E7, nt 81 to 880 with a mutant nt 226 5′ splice site [GU to GG]) (A) and pZMZ82 (16E6, nt 81 to 559 with a mutant nt 226 5′ splice site [GU to GG]), pZMZ71 (16E6*I cDNA, nt 81 to 559), pZMZ72 (16E6*II cDNA, nt 81 to 559), pZMZ73 (16E6, nt 81 to 559), and pZMZ74 (16E7, nt 560 to 880) (B). 16E6*I and 16E6*II cDNA constructs have no coding sequences in the E6 intron region because of pre-mRNA splicing. Diagrams at the top of each panel show the insertion and position of the “E6” or “E7” coding region in mammalian expression vector <t>pEGFP-C1.</t> The individual images were collected at 6 (6E6 and 11E6) or 24 h after transfection. Plasmid pEGFP-C1 was used as a GFP control in each transfection. A fluorescence image, a differential interference contrast (DIC) image, and a merged fluorescence-DIC image for each fusion protein arepresented. (A) Localization of 6E6, 11E6, 16E6, and 18E6 translated from the inserted E6E7 coding region in COS-1 cells. 16E6 mt 5′ ss indicates the plasmid containing the 16E6E7 coding region but having a mutant 5′ splice site at nt 226 position (GU to GG), used as a transfection control for full-length 16E6, since this mutation blocks the splicing of the E6E7 pre-mRNA. The single nucleotide mutation in the 5′ splice site of 16E6E7 converts a GUA codon for valine in 16E6 to a GGA codon for glycine. (B) Localization of various 16E6 proteins translated from the inserted E6 coding region and E6*I or E6*II cDNA. 16E7 protein translated from plasmid pZMZ74 was used for comparison. + or − indicates the presence or absence of cytoplasmic (C), nuclear (N), or nucleolar (No) localization of GFP fusions. Scale bars, ∼8 μm. (C) Western blotting analysis of GFP-E6 or GFP-E7 fusions expressed from pEGFP-C1 vectors. As shown in panels A and B, all GFP fusions except for mutant E6 were included for comparison of the expected full-length E6 or E7 protein, even through the 6E6 and 11E6 coding regions do not have an intron. GFP-16E6 in lane 5, encoded from a plasmid with the 16E6E7 coding region, was included for comparison with GFP-16E6 in lane 8, encoded from a plasmid with the 16E6 coding region. Protein samples prepared from COS-1 cells transfected with the various plasmids were resolved on an SDS-12% polyacrylamide gel and blotted with anti-GFP antibody. GFP and its derived fusions are indicated above the lanes. Prestained protein markers (Bio-Rad) in kilodaltons are shown at the left.
    Mammalian Expression Vectors Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    83
    TaKaRa fluorescent protein pegfp c1
    Immunocytochemical detection of MKL1 and MKL2 protein in cortical neurons with synapses. ( a ) Immunostaining of mature, cortical neuronal cultures. <t>pEGFP-C1</t> vector (4 μg/well) was transfected into cortical neurons at 18 days in culture. Three days later, cells were subjected to immunostaining using anti-GFP and anti-MKL1 or anti-MKL2 antibodies. ( b ) High magnification of GFP-positive cortical neurons at 21 days in culture. Arrowheads indicate dendritic spines. ( c , d ) Synaptic staining of MKL1 and MKL2 in mature, cortical neurons. Cortical neurons at 21 days in culture were immunostained using anti-MKL1, anti-MKL2, anti-postsynaptic density (PSD)-95, and anti-synaptophysin antibodies. MKL and PSD-95 signals merged. MKL and synaptophysin signals also merged.
    Fluorescent Protein Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 83/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa plasmid encoding enhanced green fluorescent protein pegfp c1
    GFP expressions in A10 cells incubated with <t>pEGFP-C1</t> loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).
    Plasmid Encoding Enhanced Green Fluorescent Protein Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa cloning vectors pegfp c1
    GFP expressions in A10 cells incubated with <t>pEGFP-C1</t> loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).
    Cloning Vectors Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa protein encoding plasmids pegfp c1
    GFP expressions in A10 cells incubated with <t>pEGFP-C1</t> loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).
    Protein Encoding Plasmids Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa actin pegfp c 1
    GFP expressions in A10 cells incubated with <t>pEGFP-C1</t> loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).
    Actin Pegfp C 1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa eukaryotic expression vector pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Eukaryotic Expression Vector Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 83/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa eucaryotic expression vector pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Eucaryotic Expression Vector Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa vector plasmids pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Vector Plasmids Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa fluorescent protein egfp expression vector
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Fluorescent Protein Egfp Expression Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 81/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa fluorescence protein gfp expression plasmid pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Fluorescence Protein Gfp Expression Plasmid Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa fluorescent protein gfp
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Fluorescent Protein Gfp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 1533 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    TaKaRa cytosolic gfp vector pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Cytosolic Gfp Vector Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa fluorescent protein gfp expression plasmid pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Fluorescent Protein Gfp Expression Plasmid Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa mutant gfp gene pegfp c1
    LRIG1 expression in SHG-44 cells transfected with <t>pEGFP-C1-LRIG1.</t> a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P
    Mutant Gfp Gene Pegfp C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Postmortem histology of tumors after intravenous delivery of either pCMV-TK (a, c, d) or pEGFP-C1 (b)-loaded microbubbles and treatment with ultrasound and GCV. (a, b) Acellular zones ( arrows ) visible under H E stain. (c) Merged image of two sister

    Journal: Ultrasound in medicine & biology

    Article Title: GENE THERAPY OF CARCINOMA USING ULTRASOUND-TARGETED MICROBUBBLE DESTRUCTION

    doi: 10.1016/j.ultrasmedbio.2010.11.011

    Figure Lengend Snippet: Postmortem histology of tumors after intravenous delivery of either pCMV-TK (a, c, d) or pEGFP-C1 (b)-loaded microbubbles and treatment with ultrasound and GCV. (a, b) Acellular zones ( arrows ) visible under H E stain. (c) Merged image of two sister

    Article Snippet: illustrates tumor growth after transduction of the GCV-treated mice receiving UTMD-gene delivery of pCMV-TK (TK/GCV group) or pEGFP-C1 (GFP/GCV group) from an initial size of 0.1–0.15 mL (Day 0) until the mice were euthanized.

    Techniques: Staining

    Immunofluorescent staining of GFP in murine tumors 3 d after intravenous delivery of pEGFP-C1 bound to microbubbles and treatment with ultrasound (a, b) or no ultrasound (c). GFP-positive staining was seen both in hollow structures (a) as well as individual

    Journal: Ultrasound in medicine & biology

    Article Title: GENE THERAPY OF CARCINOMA USING ULTRASOUND-TARGETED MICROBUBBLE DESTRUCTION

    doi: 10.1016/j.ultrasmedbio.2010.11.011

    Figure Lengend Snippet: Immunofluorescent staining of GFP in murine tumors 3 d after intravenous delivery of pEGFP-C1 bound to microbubbles and treatment with ultrasound (a, b) or no ultrasound (c). GFP-positive staining was seen both in hollow structures (a) as well as individual

    Article Snippet: illustrates tumor growth after transduction of the GCV-treated mice receiving UTMD-gene delivery of pCMV-TK (TK/GCV group) or pEGFP-C1 (GFP/GCV group) from an initial size of 0.1–0.15 mL (Day 0) until the mice were euthanized.

    Techniques: Staining

    Postmortem TUNEL assays from murine tumors after intravenous injection of either pCMV-TK (a) or pEGFP-C1 (b)-loaded microbubbles and treatment with ultrasound and GCV.

    Journal: Ultrasound in medicine & biology

    Article Title: GENE THERAPY OF CARCINOMA USING ULTRASOUND-TARGETED MICROBUBBLE DESTRUCTION

    doi: 10.1016/j.ultrasmedbio.2010.11.011

    Figure Lengend Snippet: Postmortem TUNEL assays from murine tumors after intravenous injection of either pCMV-TK (a) or pEGFP-C1 (b)-loaded microbubbles and treatment with ultrasound and GCV.

    Article Snippet: illustrates tumor growth after transduction of the GCV-treated mice receiving UTMD-gene delivery of pCMV-TK (TK/GCV group) or pEGFP-C1 (GFP/GCV group) from an initial size of 0.1–0.15 mL (Day 0) until the mice were euthanized.

    Techniques: TUNEL Assay, Injection

    Growth of murine tumors after intravenous injection of either pCMV-TK (□) or pEGFP-C1 (▲)-loaded microbubbles and treated with ultrasound. Daily GCV injections began on day 3. Best-fit lines were calculated from all data points in each

    Journal: Ultrasound in medicine & biology

    Article Title: GENE THERAPY OF CARCINOMA USING ULTRASOUND-TARGETED MICROBUBBLE DESTRUCTION

    doi: 10.1016/j.ultrasmedbio.2010.11.011

    Figure Lengend Snippet: Growth of murine tumors after intravenous injection of either pCMV-TK (□) or pEGFP-C1 (▲)-loaded microbubbles and treated with ultrasound. Daily GCV injections began on day 3. Best-fit lines were calculated from all data points in each

    Article Snippet: illustrates tumor growth after transduction of the GCV-treated mice receiving UTMD-gene delivery of pCMV-TK (TK/GCV group) or pEGFP-C1 (GFP/GCV group) from an initial size of 0.1–0.15 mL (Day 0) until the mice were euthanized.

    Techniques: Injection

    Rab5 and Rabankyrin5 overexpression increases macropinosome formation. The distribution of Rab5, Rabankyrin-5 (A) or Rab5(Q79L) (B) in green relative to dextran-positive macropinosomes (red) Scale bar = 10 µm. (C) HEK-Flp-In cells were transiently transfected with pEGFP-C1, pEGFP-Rab5, pEGFP-Rab5-Q79L and pEYFP-Rabankyrin-5. The mean number of macropinosomes/100 transfected cells was quantitated over 3 replicates of 500 transfected cells for each condition. * denotes statistical significance (p

    Journal: PLoS ONE

    Article Title: The SNX-PX-BAR Family in Macropinocytosis: The Regulation of Macropinosome Formation by SNX-PX-BAR Proteins

    doi: 10.1371/journal.pone.0013763

    Figure Lengend Snippet: Rab5 and Rabankyrin5 overexpression increases macropinosome formation. The distribution of Rab5, Rabankyrin-5 (A) or Rab5(Q79L) (B) in green relative to dextran-positive macropinosomes (red) Scale bar = 10 µm. (C) HEK-Flp-In cells were transiently transfected with pEGFP-C1, pEGFP-Rab5, pEGFP-Rab5-Q79L and pEYFP-Rabankyrin-5. The mean number of macropinosomes/100 transfected cells was quantitated over 3 replicates of 500 transfected cells for each condition. * denotes statistical significance (p

    Article Snippet: The construct pEGFP-C1 was obtained from Clontech. pEGFP-Rab5 , pEGFP-Rab5Q79L , and pEYFP-Rabankyrin 5 , are as described previously. pIRES2-EGFP-PTEN and pIRES2-EGFP-PTEN(G129E) were generated by subcloning the open-reading frames of PTEN and PTEN(G129E) into pIRES2-EGFP using BamHI and EcoRI. pEGFP-SNX1, pEGFP-SNX2, pEGFP-SNX4, pEGFP-SNX5, pEGFP-SNX6, pEGFP-SNX7, pEGFP-SNX8, pEGFP-SNX9, pEGFP-SNX18, pEGFP-SNX30, pEGFP-SNX32, and pEGFP-SNX33 were generated by amplifying their full length open reading frame by polymerase chain reaction (PCR) and the resulting PCR products cloned into pEGFP-C1. pmCherry-SNX18 was generated by amplifying the full length open reading frame of SNX18 and cloning the resulting PCR product into pmCherry-C1. pEGFP-ΔSH3-SNX18 was generated by amplifying residues 61-615 of SNX18 and cloning the resulting PCR product into pEGFP-C1.

    Techniques: Over Expression, Transfection

    Macropinosome formation screening assay validation. A: 24 hours post transfection, HEK-Flp-In cell monolayers were pulsed for 5 minutes with 100 µg/mL dextran (10,000 MW) conjugated to tetramethylrhodamine (dextran-TR) at 37°C. The samples were then washed in 4°C PBS, fixed in 4% PFA and imaged and processed as described in Materials and Methods . Briefly Z-stack images comprising of 3×5 µm Z slices were merged into a single RGB image of transfected cells (green) stained with dextran-TR (red) (Ai). The red channel from the RGB image was isolated and converted to an 8-bit grayscale image (Aii). Dextran-positive macropinosomes were selected based on size ( > 0.5 µm in diameter) and fluorescent intensity ( > 100). Selected macropinosomes are shown in the foreground as black (Aiii). This binary image was then converted to a mask and superimposed onto the Green channel of the original RGB image to measure the green fluorescent intensity of the area occupied by each macropinosome in the image (Aiv). The particles with green fluorescence intensity higher than background signal ( > 20) were considered to be macropinosomes within a transfected cell, represented in green. Red particles represent discarded macropinosomes determined to be outside of a transfected cell. Scale = 10 µm. B, C, D: HEK-Flp-In cell monolayers were either serum-starved for 16 hours and treated with dextran-TR in the presence or absence of 100 ng/mL EGF for 5 minutes at 37°C (B), treated with 1 mM amiloride or carrier (0.6% Methanol) for 30 minutes at 37°C before pulsing with dextran-TR (C), or transiently transfected with pEGFP-C1 or pEGFP-SNX5 before pulsing with dextran-TR (D). The samples in each case were assayed for macropinosome formation as described in Materials and Methods , quantitating the mean number of macropinosomes/100 transfected cells over 3 replicates of 500 transfected cells for each condition. * denotes statistical significance (p

    Journal: PLoS ONE

    Article Title: The SNX-PX-BAR Family in Macropinocytosis: The Regulation of Macropinosome Formation by SNX-PX-BAR Proteins

    doi: 10.1371/journal.pone.0013763

    Figure Lengend Snippet: Macropinosome formation screening assay validation. A: 24 hours post transfection, HEK-Flp-In cell monolayers were pulsed for 5 minutes with 100 µg/mL dextran (10,000 MW) conjugated to tetramethylrhodamine (dextran-TR) at 37°C. The samples were then washed in 4°C PBS, fixed in 4% PFA and imaged and processed as described in Materials and Methods . Briefly Z-stack images comprising of 3×5 µm Z slices were merged into a single RGB image of transfected cells (green) stained with dextran-TR (red) (Ai). The red channel from the RGB image was isolated and converted to an 8-bit grayscale image (Aii). Dextran-positive macropinosomes were selected based on size ( > 0.5 µm in diameter) and fluorescent intensity ( > 100). Selected macropinosomes are shown in the foreground as black (Aiii). This binary image was then converted to a mask and superimposed onto the Green channel of the original RGB image to measure the green fluorescent intensity of the area occupied by each macropinosome in the image (Aiv). The particles with green fluorescence intensity higher than background signal ( > 20) were considered to be macropinosomes within a transfected cell, represented in green. Red particles represent discarded macropinosomes determined to be outside of a transfected cell. Scale = 10 µm. B, C, D: HEK-Flp-In cell monolayers were either serum-starved for 16 hours and treated with dextran-TR in the presence or absence of 100 ng/mL EGF for 5 minutes at 37°C (B), treated with 1 mM amiloride or carrier (0.6% Methanol) for 30 minutes at 37°C before pulsing with dextran-TR (C), or transiently transfected with pEGFP-C1 or pEGFP-SNX5 before pulsing with dextran-TR (D). The samples in each case were assayed for macropinosome formation as described in Materials and Methods , quantitating the mean number of macropinosomes/100 transfected cells over 3 replicates of 500 transfected cells for each condition. * denotes statistical significance (p

    Article Snippet: The construct pEGFP-C1 was obtained from Clontech. pEGFP-Rab5 , pEGFP-Rab5Q79L , and pEYFP-Rabankyrin 5 , are as described previously. pIRES2-EGFP-PTEN and pIRES2-EGFP-PTEN(G129E) were generated by subcloning the open-reading frames of PTEN and PTEN(G129E) into pIRES2-EGFP using BamHI and EcoRI. pEGFP-SNX1, pEGFP-SNX2, pEGFP-SNX4, pEGFP-SNX5, pEGFP-SNX6, pEGFP-SNX7, pEGFP-SNX8, pEGFP-SNX9, pEGFP-SNX18, pEGFP-SNX30, pEGFP-SNX32, and pEGFP-SNX33 were generated by amplifying their full length open reading frame by polymerase chain reaction (PCR) and the resulting PCR products cloned into pEGFP-C1. pmCherry-SNX18 was generated by amplifying the full length open reading frame of SNX18 and cloning the resulting PCR product into pmCherry-C1. pEGFP-ΔSH3-SNX18 was generated by amplifying residues 61-615 of SNX18 and cloning the resulting PCR product into pEGFP-C1.

    Techniques: Screening Assay, Transfection, Staining, Isolation, Fluorescence

    Schematic representation of vectors for evaluating the impact of different MARs combination on recombinant protein expression level and stability in CHO cells ( A ). Map of pEGFP-C1 used in this study ( B ). SpA, simian virus 40 early polyadenylation signal; eGFP, enhanced green fluorescence protein.

    Journal: Scientific Reports

    Article Title: Matrix attachment region combinations increase transgene expression in transfected Chinese hamster ovary cells

    doi: 10.1038/srep42805

    Figure Lengend Snippet: Schematic representation of vectors for evaluating the impact of different MARs combination on recombinant protein expression level and stability in CHO cells ( A ). Map of pEGFP-C1 used in this study ( B ). SpA, simian virus 40 early polyadenylation signal; eGFP, enhanced green fluorescence protein.

    Article Snippet: Plasmids and constructs pEGFP-C1 containing the CMV promoter was used as the backbone vector in this study (Clontech, Mountain View, CA, USA).

    Techniques: Recombinant, Expressing, Fluorescence

    Confocal microscopic analysis of M2 expression. A20 cells were transfected with either pEGFP-C1 (A to C) or pEGFP-C1/M2 (D to I) and after 48 h were either fixed, permeabilized, and then counterstained with PI or stained with TRITC-conjugated antibody to MHC class II antigens and then fixed. Cells were then visualized using a Leica TCNTS confocal microscope with combinations of laser light wavelength and band filter specific for EGFP (488 and 525/50 nm [A, D, and G]) or for PI and TRITC (568 and 600/30 nm [B, E, and H]). Image analysis with Leica software enabled the overlaying of these two images (C, F, and I). All images are shown at the same magnification.

    Journal: Journal of Virology

    Article Title: Murid Herpesvirus 4 Strain 68 M2 Protein Is a B-Cell-Associated Antigen Important for Latency but Not Lymphocytosis

    doi: 10.1128/JVI.77.17.9700-9709.2003

    Figure Lengend Snippet: Confocal microscopic analysis of M2 expression. A20 cells were transfected with either pEGFP-C1 (A to C) or pEGFP-C1/M2 (D to I) and after 48 h were either fixed, permeabilized, and then counterstained with PI or stained with TRITC-conjugated antibody to MHC class II antigens and then fixed. Cells were then visualized using a Leica TCNTS confocal microscope with combinations of laser light wavelength and band filter specific for EGFP (488 and 525/50 nm [A, D, and G]) or for PI and TRITC (568 and 600/30 nm [B, E, and H]). Image analysis with Leica software enabled the overlaying of these two images (C, F, and I). All images are shown at the same magnification.

    Article Snippet: Because of the lack of an adequate M2-specific antibody, M2 was expressed as either an N- or C-terminal fusion with enhanced green fluorescent protein (EGFP) by using the vectors pEGFP-C1 and pEGFP-N1 (Clontech) to aid in its detection.

    Techniques: Expressing, Transfection, Staining, Microscopy, Software

    Green fluorescent protein (GFP) expression in C17.2 neural stem cells after transfection. (A) C17.2 neural stem cells were transfected with 0.6, 1.2, 1.8, 2.4 or 3.0 μg pEGFP expression plasmid. 1.2 μg of pEGFP displayed the most effective transfection rate of more than 85% at different time points post-transfection. Results were assessed using GFP fluorescence intensity. (B, C) C17.2 neural stem cells abundantly expressed the GFP protein (green) 24 hours after transfection (visualized under an inverted fluorescence microscope). (B) Scale bar: 200 μm; (C) scale bar: 50 μm.

    Journal: Neural Regeneration Research

    Article Title: Targeting β-secretase with RNAi in neural stem cells for Alzheimer's disease therapy

    doi: 10.3969/j.issn.1673-5374.2013.33.003

    Figure Lengend Snippet: Green fluorescent protein (GFP) expression in C17.2 neural stem cells after transfection. (A) C17.2 neural stem cells were transfected with 0.6, 1.2, 1.8, 2.4 or 3.0 μg pEGFP expression plasmid. 1.2 μg of pEGFP displayed the most effective transfection rate of more than 85% at different time points post-transfection. Results were assessed using GFP fluorescence intensity. (B, C) C17.2 neural stem cells abundantly expressed the GFP protein (green) 24 hours after transfection (visualized under an inverted fluorescence microscope). (B) Scale bar: 200 μm; (C) scale bar: 50 μm.

    Article Snippet: After about 80% confluence, the GFP-expressing plasmid (pEGFP-C1) and psiEGFP plasmid (Clontech, Mountain View, CA, USA) encoding the shRNA against the EGFP gene were simultaneously transfected into neural stem cells to facilitate the evaluation of the effectiveness of the exogenously introduced shRNA.

    Techniques: Expressing, Transfection, Plasmid Preparation, Fluorescence, Microscopy

    MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid pEGFP C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Rab27a Is an Essential Component of Melanosome Receptor for Myosin Va

    doi: 10.1091/mbc.01-12-0595

    Figure Lengend Snippet: MC MV and MC MVΔD, but not MC MVΔF and BR MV, rescue melanosome distribution in dilute melanocytes. Shown is the distribution of melanosomes in brightfield (A, D, G, and J), the cell shape in phase contrast (B, E, H, and K), and the distribution of GFP fluorescence due primarily to unfused GFP expressed from plasmid pEGFP C1 (C, F, I, and L), which was coinjected to ensure accurate identification of all injected cells (see text), in dilute melanocytes microinjected with MC MV (A–C), MC MVΔD (D–F), MC MVΔF (G–I), or BR MV (J–L). Bars, 11.5 μm.

    Article Snippet: The resulting fragment was digested with Bam HI and Eco RI and cloned into plasmid pEGFP C1 that had been cut with Bgl II and Eco RI.

    Techniques: Fluorescence, Plasmid Preparation, Injection

    Overexpressed GJA8 protein modulated the level of LC3 and P62 in HLE cells. a HLE cells stained for GJA8 (red) and LC3 (green) in the NC and Sta2h groups. b Western blot with LC3-I/II in NC and Sta2h groups in HLE cells. c The average level of LC3-II/Actin of three independent experiments. d Transfected with peGFP-C1-GJA8 in the NC and Sta2h groups, HLE cells stained for LC3 (red). e Mean number of LC3 puncta for each treatment ( n = 3 wells, 3 independent experiments, > 50 cells per experiment). f HLE cells for corresponding treatment and transfected with peGFP-C1-GJA8 for 0ug or 1ug or 2ug, testing the expression of P62 and α-Tubulin. g The average level of P62/α-Tubulin of three independent experiments in Sta4h/Sta2h/NC group. h The average level of P62/α-Tubulin of three independent experiments transfected with peGFP-C1-GJA8 for 0ug or 1ug or 2ug in NC group. All values are represented as the mean + SEM; * p

    Journal: Human Genetics

    Article Title: The impact of GJA8 SNPs on susceptibility to age-related cataract

    doi: 10.1007/s00439-018-1945-5

    Figure Lengend Snippet: Overexpressed GJA8 protein modulated the level of LC3 and P62 in HLE cells. a HLE cells stained for GJA8 (red) and LC3 (green) in the NC and Sta2h groups. b Western blot with LC3-I/II in NC and Sta2h groups in HLE cells. c The average level of LC3-II/Actin of three independent experiments. d Transfected with peGFP-C1-GJA8 in the NC and Sta2h groups, HLE cells stained for LC3 (red). e Mean number of LC3 puncta for each treatment ( n = 3 wells, 3 independent experiments, > 50 cells per experiment). f HLE cells for corresponding treatment and transfected with peGFP-C1-GJA8 for 0ug or 1ug or 2ug, testing the expression of P62 and α-Tubulin. g The average level of P62/α-Tubulin of three independent experiments in Sta4h/Sta2h/NC group. h The average level of P62/α-Tubulin of three independent experiments transfected with peGFP-C1-GJA8 for 0ug or 1ug or 2ug in NC group. All values are represented as the mean + SEM; * p

    Article Snippet: The PCR products and vector pEGFP-C1 were digested by Xho I and Eco rI (Takara, Japan).

    Techniques: Staining, Western Blot, Transfection, Expressing

    NS6 interacts with both RIG-I and MDA5. (A to D) HEK-293T cells were cotransfected with pCAGGS-HA-NS6 and Flag-tagged RIG-I (A and C) or Flag-tagged MDA5 (B and D), respectively. At 28 h after transfection, cells were lysed and subjected to immunoprecipitation analysis with anti-HA (IP: HA) or anti-Flag (IP: Flag) antibody. The whole-cell lysates (WCL) and immunoprecipitation (IP) complexes were analyzed via Western blotting using anti-Flag, anti-HA, or anti-β-actin antibodies. (E and F) HEK-293T cells were cotransfected with pCAGGS-HA-NS6 and Flag-tagged RIG-I (E) or MDA5 (F). At 28 h after transfection, the cells were fixed for immunofluorescence assays to detect NS6 protein (red) and RIG-I or MDA5 (green) with anti-HA and anti-Flag antibodies, respectively. (G and H) HEK-293T cells were transfected with pCAGGS-HA-NS6 and pEGFP-C1, Flag-tagged RIG-I (G), or MDA5 (H) expression plasmids for 24 h, followed by the transfection of poly(I·C). Cells were lysed 36 h after transfection, and the clarified supernatants were left untreated or were treated with RNase A (50 μg/ml). The samples were then subjected to immunoprecipitation assays using anti-HA MAb (IP: HA). Cell lysates and immunoprecipitated complexes were subjected to Western blot assays as described for panels A and B.

    Journal: Journal of Virology

    Article Title: Porcine Deltacoronavirus Accessory Protein NS6 Antagonizes Interferon Beta Production by Interfering with the Binding of RIG-I/MDA5 to Double-Stranded RNA

    doi: 10.1128/JVI.00712-18

    Figure Lengend Snippet: NS6 interacts with both RIG-I and MDA5. (A to D) HEK-293T cells were cotransfected with pCAGGS-HA-NS6 and Flag-tagged RIG-I (A and C) or Flag-tagged MDA5 (B and D), respectively. At 28 h after transfection, cells were lysed and subjected to immunoprecipitation analysis with anti-HA (IP: HA) or anti-Flag (IP: Flag) antibody. The whole-cell lysates (WCL) and immunoprecipitation (IP) complexes were analyzed via Western blotting using anti-Flag, anti-HA, or anti-β-actin antibodies. (E and F) HEK-293T cells were cotransfected with pCAGGS-HA-NS6 and Flag-tagged RIG-I (E) or MDA5 (F). At 28 h after transfection, the cells were fixed for immunofluorescence assays to detect NS6 protein (red) and RIG-I or MDA5 (green) with anti-HA and anti-Flag antibodies, respectively. (G and H) HEK-293T cells were transfected with pCAGGS-HA-NS6 and pEGFP-C1, Flag-tagged RIG-I (G), or MDA5 (H) expression plasmids for 24 h, followed by the transfection of poly(I·C). Cells were lysed 36 h after transfection, and the clarified supernatants were left untreated or were treated with RNase A (50 μg/ml). The samples were then subjected to immunoprecipitation assays using anti-HA MAb (IP: HA). Cell lysates and immunoprecipitated complexes were subjected to Western blot assays as described for panels A and B.

    Article Snippet: The GFP expression plasmid (pEGFP-C1) was purchased from TaKaRa (Japan).

    Techniques: Transfection, Immunoprecipitation, Western Blot, Immunofluorescence, Expressing

    Localization of MT and Impα fusion proteins. HeLa S3 cells were transfected with pEGFP-C1–MT and pDsRed1-N1–Impα ( A–C ) or pEGFP-C1–MT (144–476) ( D ), constructed as recommended by Clontech, and analyzed by fluorescence microscopy.

    Journal: Genes & Development

    Article Title: Cap methyltransferase selective binding and methylation of GpppG-RNA are stimulated by importin-?

    doi:

    Figure Lengend Snippet: Localization of MT and Impα fusion proteins. HeLa S3 cells were transfected with pEGFP-C1–MT and pDsRed1-N1–Impα ( A–C ) or pEGFP-C1–MT (144–476) ( D ), constructed as recommended by Clontech, and analyzed by fluorescence microscopy.

    Article Snippet: MT and MT (144–476) were ligated into GFP fusion vector pEGFP-C1, and Impα was cloned into RFP fusion vector pDsRed1-N1 (Clontech).

    Techniques: Transfection, Construct, Fluorescence, Microscopy

    MT/Impα binding. Purified GST and GST–MT ( A ) or GST–Impα ( B ) were incubated with [ 35 ). HeLa S3 cells ( C ) were transfected with pEGFP-C1–Impα and pcDNA3.1(+)-MT-myc. Two days later, cell lysates were immunoprecipitated (IP) in RIPA buffer (see Materials and Methods) as indicated, with anti-myc (α-myc) or anti-GFP (α-GFP) antibodies, and immunoblotted with α-GFP or α-myc. (G) GFP only; (G-I) GFP–Impα fusion protein; (m) myc tag only; (M-m) MT–myc fusion protein.

    Journal: Genes & Development

    Article Title: Cap methyltransferase selective binding and methylation of GpppG-RNA are stimulated by importin-?

    doi:

    Figure Lengend Snippet: MT/Impα binding. Purified GST and GST–MT ( A ) or GST–Impα ( B ) were incubated with [ 35 ). HeLa S3 cells ( C ) were transfected with pEGFP-C1–Impα and pcDNA3.1(+)-MT-myc. Two days later, cell lysates were immunoprecipitated (IP) in RIPA buffer (see Materials and Methods) as indicated, with anti-myc (α-myc) or anti-GFP (α-GFP) antibodies, and immunoblotted with α-GFP or α-myc. (G) GFP only; (G-I) GFP–Impα fusion protein; (m) myc tag only; (M-m) MT–myc fusion protein.

    Article Snippet: MT and MT (144–476) were ligated into GFP fusion vector pEGFP-C1, and Impα was cloned into RFP fusion vector pDsRed1-N1 (Clontech).

    Techniques: Binding Assay, Purification, Incubation, Transfection, Immunoprecipitation

    Cellular localizations of low-risk and high-risk HPV E6 in COS-1 cells. COS-1 cells were transfected with plasmids pZMZ66 (6E6E7, nt 85 to 939), pZMZ67 (11E6E7, nt 85 to 924), pZMZ69 (18E6E7, nt 103 to 967), pZMZ70 (16E6E7, nt 81 to 880), and pZMZ81 (16E6E7, nt 81 to 880 with a mutant nt 226 5′ splice site [GU to GG]) (A) and pZMZ82 (16E6, nt 81 to 559 with a mutant nt 226 5′ splice site [GU to GG]), pZMZ71 (16E6*I cDNA, nt 81 to 559), pZMZ72 (16E6*II cDNA, nt 81 to 559), pZMZ73 (16E6, nt 81 to 559), and pZMZ74 (16E7, nt 560 to 880) (B). 16E6*I and 16E6*II cDNA constructs have no coding sequences in the E6 intron region because of pre-mRNA splicing. Diagrams at the top of each panel show the insertion and position of the “E6” or “E7” coding region in mammalian expression vector pEGFP-C1. The individual images were collected at 6 (6E6 and 11E6) or 24 h after transfection. Plasmid pEGFP-C1 was used as a GFP control in each transfection. A fluorescence image, a differential interference contrast (DIC) image, and a merged fluorescence-DIC image for each fusion protein arepresented. (A) Localization of 6E6, 11E6, 16E6, and 18E6 translated from the inserted E6E7 coding region in COS-1 cells. 16E6 mt 5′ ss indicates the plasmid containing the 16E6E7 coding region but having a mutant 5′ splice site at nt 226 position (GU to GG), used as a transfection control for full-length 16E6, since this mutation blocks the splicing of the E6E7 pre-mRNA. The single nucleotide mutation in the 5′ splice site of 16E6E7 converts a GUA codon for valine in 16E6 to a GGA codon for glycine. (B) Localization of various 16E6 proteins translated from the inserted E6 coding region and E6*I or E6*II cDNA. 16E7 protein translated from plasmid pZMZ74 was used for comparison. + or − indicates the presence or absence of cytoplasmic (C), nuclear (N), or nucleolar (No) localization of GFP fusions. Scale bars, ∼8 μm. (C) Western blotting analysis of GFP-E6 or GFP-E7 fusions expressed from pEGFP-C1 vectors. As shown in panels A and B, all GFP fusions except for mutant E6 were included for comparison of the expected full-length E6 or E7 protein, even through the 6E6 and 11E6 coding regions do not have an intron. GFP-16E6 in lane 5, encoded from a plasmid with the 16E6E7 coding region, was included for comparison with GFP-16E6 in lane 8, encoded from a plasmid with the 16E6 coding region. Protein samples prepared from COS-1 cells transfected with the various plasmids were resolved on an SDS-12% polyacrylamide gel and blotted with anti-GFP antibody. GFP and its derived fusions are indicated above the lanes. Prestained protein markers (Bio-Rad) in kilodaltons are shown at the left.

    Journal: Journal of Virology

    Article Title: Signals That Dictate Nuclear Localization of Human Papillomavirus Type 16 Oncoprotein E6 in Living Cells

    doi: 10.1128/JVI.77.24.13232-13247.2003

    Figure Lengend Snippet: Cellular localizations of low-risk and high-risk HPV E6 in COS-1 cells. COS-1 cells were transfected with plasmids pZMZ66 (6E6E7, nt 85 to 939), pZMZ67 (11E6E7, nt 85 to 924), pZMZ69 (18E6E7, nt 103 to 967), pZMZ70 (16E6E7, nt 81 to 880), and pZMZ81 (16E6E7, nt 81 to 880 with a mutant nt 226 5′ splice site [GU to GG]) (A) and pZMZ82 (16E6, nt 81 to 559 with a mutant nt 226 5′ splice site [GU to GG]), pZMZ71 (16E6*I cDNA, nt 81 to 559), pZMZ72 (16E6*II cDNA, nt 81 to 559), pZMZ73 (16E6, nt 81 to 559), and pZMZ74 (16E7, nt 560 to 880) (B). 16E6*I and 16E6*II cDNA constructs have no coding sequences in the E6 intron region because of pre-mRNA splicing. Diagrams at the top of each panel show the insertion and position of the “E6” or “E7” coding region in mammalian expression vector pEGFP-C1. The individual images were collected at 6 (6E6 and 11E6) or 24 h after transfection. Plasmid pEGFP-C1 was used as a GFP control in each transfection. A fluorescence image, a differential interference contrast (DIC) image, and a merged fluorescence-DIC image for each fusion protein arepresented. (A) Localization of 6E6, 11E6, 16E6, and 18E6 translated from the inserted E6E7 coding region in COS-1 cells. 16E6 mt 5′ ss indicates the plasmid containing the 16E6E7 coding region but having a mutant 5′ splice site at nt 226 position (GU to GG), used as a transfection control for full-length 16E6, since this mutation blocks the splicing of the E6E7 pre-mRNA. The single nucleotide mutation in the 5′ splice site of 16E6E7 converts a GUA codon for valine in 16E6 to a GGA codon for glycine. (B) Localization of various 16E6 proteins translated from the inserted E6 coding region and E6*I or E6*II cDNA. 16E7 protein translated from plasmid pZMZ74 was used for comparison. + or − indicates the presence or absence of cytoplasmic (C), nuclear (N), or nucleolar (No) localization of GFP fusions. Scale bars, ∼8 μm. (C) Western blotting analysis of GFP-E6 or GFP-E7 fusions expressed from pEGFP-C1 vectors. As shown in panels A and B, all GFP fusions except for mutant E6 were included for comparison of the expected full-length E6 or E7 protein, even through the 6E6 and 11E6 coding regions do not have an intron. GFP-16E6 in lane 5, encoded from a plasmid with the 16E6E7 coding region, was included for comparison with GFP-16E6 in lane 8, encoded from a plasmid with the 16E6 coding region. Protein samples prepared from COS-1 cells transfected with the various plasmids were resolved on an SDS-12% polyacrylamide gel and blotted with anti-GFP antibody. GFP and its derived fusions are indicated above the lanes. Prestained protein markers (Bio-Rad) in kilodaltons are shown at the left.

    Article Snippet: Mammalian expression vectors pEGFP-C1 and pEGFP-N1 were purchased from Clontech (Palo Alto, Calif.).

    Techniques: Transfection, Mutagenesis, Construct, Expressing, Plasmid Preparation, Fluorescence, Western Blot, Derivative Assay

    Immunocytochemical detection of MKL1 and MKL2 protein in cortical neurons with synapses. ( a ) Immunostaining of mature, cortical neuronal cultures. pEGFP-C1 vector (4 μg/well) was transfected into cortical neurons at 18 days in culture. Three days later, cells were subjected to immunostaining using anti-GFP and anti-MKL1 or anti-MKL2 antibodies. ( b ) High magnification of GFP-positive cortical neurons at 21 days in culture. Arrowheads indicate dendritic spines. ( c , d ) Synaptic staining of MKL1 and MKL2 in mature, cortical neurons. Cortical neurons at 21 days in culture were immunostained using anti-MKL1, anti-MKL2, anti-postsynaptic density (PSD)-95, and anti-synaptophysin antibodies. MKL and PSD-95 signals merged. MKL and synaptophysin signals also merged.

    Journal: Scientific Reports

    Article Title: Synaptic localisation of SRF coactivators, MKL1 and MKL2, and their role in dendritic spine morphology

    doi: 10.1038/s41598-017-18905-7

    Figure Lengend Snippet: Immunocytochemical detection of MKL1 and MKL2 protein in cortical neurons with synapses. ( a ) Immunostaining of mature, cortical neuronal cultures. pEGFP-C1 vector (4 μg/well) was transfected into cortical neurons at 18 days in culture. Three days later, cells were subjected to immunostaining using anti-GFP and anti-MKL1 or anti-MKL2 antibodies. ( b ) High magnification of GFP-positive cortical neurons at 21 days in culture. Arrowheads indicate dendritic spines. ( c , d ) Synaptic staining of MKL1 and MKL2 in mature, cortical neurons. Cortical neurons at 21 days in culture were immunostained using anti-MKL1, anti-MKL2, anti-postsynaptic density (PSD)-95, and anti-synaptophysin antibodies. MKL and PSD-95 signals merged. MKL and synaptophysin signals also merged.

    Article Snippet: The expression vector for enhanced green fluorescent protein (pEGFP-C1) was purchased from Clontech.

    Techniques: Immunostaining, Plasmid Preparation, Transfection, Staining

    Effect of MKL1 or MKL2 knock-down on dendritic spine morphology and density in cortical neurons. ( a ) Spine morphology of cortical neurons transfected with pEGFP-C1 (1 μg/well) and shR-luc, sh MKL1 , or sh MKL2 (1 μg/well). Transfection was performed at 16 days in culture. Neurons (21 days in culture) were immunostained using anti-GFP antibody. ( b , c ) Spine morphology (m and s, mushroom or stubby; t and f, thin or filopodia; irregular) and spine density. Graphs show mean ± S.D. from seven independent experiments. The statistical significance of differences (vs. shR-luc) was analysed by ANOVA with the Tukey–Kramer test. * p

    Journal: Scientific Reports

    Article Title: Synaptic localisation of SRF coactivators, MKL1 and MKL2, and their role in dendritic spine morphology

    doi: 10.1038/s41598-017-18905-7

    Figure Lengend Snippet: Effect of MKL1 or MKL2 knock-down on dendritic spine morphology and density in cortical neurons. ( a ) Spine morphology of cortical neurons transfected with pEGFP-C1 (1 μg/well) and shR-luc, sh MKL1 , or sh MKL2 (1 μg/well). Transfection was performed at 16 days in culture. Neurons (21 days in culture) were immunostained using anti-GFP antibody. ( b , c ) Spine morphology (m and s, mushroom or stubby; t and f, thin or filopodia; irregular) and spine density. Graphs show mean ± S.D. from seven independent experiments. The statistical significance of differences (vs. shR-luc) was analysed by ANOVA with the Tukey–Kramer test. * p

    Article Snippet: The expression vector for enhanced green fluorescent protein (pEGFP-C1) was purchased from Clontech.

    Techniques: Transfection

    Specific detection of MKL1 and MKL2 by anti-MKL1 and anti-MKL2 antibodies. ( a ) pEGFP-C1 vector and pFLAG-CMV2 vector (FLAG-empty) or pFLAG-CMV2-MKL1 vector (FLAG-MKL1), pCMV-HA vector (HA-empty) or pCMV-HA-MKL2 vector (HA-MKL2) were co-transfected into NIH3T3 cells. Cell lysates were lysed 24 hours after transfection. Anti-MKL1 (MKL1) and anti-MKL2 (MKL2) antibody were used for detection of exogenous and endogenous MKL1 and MKL2, respectively. Anti-FLAG and anti-HA antibodies were used for detection of exogenous MKL1 and MKL2, respectively. Anti-α-tubulin and anti-GFP antibodies were used as internal (loading) controls. ( b ) Detection of endogenous MKL1 and MKL2 in cortical neurons. Left-most lanes: cell lysates from NIH3T3 cells overexpressing FLAG-MKL1 and HA-MKL2. Lane 2: cell lysates from untransfected NIH3T3 cells. Lane 3: cell lysates from rat cortical neurons at 7 days in culture. Full-length blots are presented in Supplementary Figure S5 .

    Journal: Scientific Reports

    Article Title: Synaptic localisation of SRF coactivators, MKL1 and MKL2, and their role in dendritic spine morphology

    doi: 10.1038/s41598-017-18905-7

    Figure Lengend Snippet: Specific detection of MKL1 and MKL2 by anti-MKL1 and anti-MKL2 antibodies. ( a ) pEGFP-C1 vector and pFLAG-CMV2 vector (FLAG-empty) or pFLAG-CMV2-MKL1 vector (FLAG-MKL1), pCMV-HA vector (HA-empty) or pCMV-HA-MKL2 vector (HA-MKL2) were co-transfected into NIH3T3 cells. Cell lysates were lysed 24 hours after transfection. Anti-MKL1 (MKL1) and anti-MKL2 (MKL2) antibody were used for detection of exogenous and endogenous MKL1 and MKL2, respectively. Anti-FLAG and anti-HA antibodies were used for detection of exogenous MKL1 and MKL2, respectively. Anti-α-tubulin and anti-GFP antibodies were used as internal (loading) controls. ( b ) Detection of endogenous MKL1 and MKL2 in cortical neurons. Left-most lanes: cell lysates from NIH3T3 cells overexpressing FLAG-MKL1 and HA-MKL2. Lane 2: cell lysates from untransfected NIH3T3 cells. Lane 3: cell lysates from rat cortical neurons at 7 days in culture. Full-length blots are presented in Supplementary Figure S5 .

    Article Snippet: The expression vector for enhanced green fluorescent protein (pEGFP-C1) was purchased from Clontech.

    Techniques: Plasmid Preparation, Transfection

    GFP expressions in A10 cells incubated with pEGFP-C1 loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).

    Journal: International Journal of Nanomedicine

    Article Title: Local gene delivery via endovascular stents coated with dodecylated chitosan-plasmid DNA nanoparticles

    doi: 10.2147/IJN.S14358

    Figure Lengend Snippet: GFP expressions in A10 cells incubated with pEGFP-C1 loaded NP-stents. A ) pEGFP-C1-loaded NP-stents; B ) dodecylated chitosan-coating stent (no plasmid DNA) (FITC, 100×).

    Article Snippet: Plasmid encoding enhanced green fluorescent protein (pEGFP-C1) was purchased from Clontech (Palo Alto, CA).

    Techniques: Incubation, Plasmid Preparation

    Rabbit CCA results demonstrating cells expressing GFP mediated by pEGFP-C1-loaded NP stents in vivo.

    Journal: International Journal of Nanomedicine

    Article Title: Local gene delivery via endovascular stents coated with dodecylated chitosan-plasmid DNA nanoparticles

    doi: 10.2147/IJN.S14358

    Figure Lengend Snippet: Rabbit CCA results demonstrating cells expressing GFP mediated by pEGFP-C1-loaded NP stents in vivo.

    Article Snippet: Plasmid encoding enhanced green fluorescent protein (pEGFP-C1) was purchased from Clontech (Palo Alto, CA).

    Techniques: Expressing, In Vivo

    LRIG1 expression in SHG-44 cells transfected with pEGFP-C1-LRIG1. a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P

    Journal: Cell Stress & Chaperones

    Article Title: LRIG1 inhibits hypoxia-induced vasculogenic mimicry formation via suppression of the EGFR/PI3K/AKT pathway and epithelial-to-mesenchymal transition in human glioma SHG-44 cells

    doi: 10.1007/s12192-015-0587-y

    Figure Lengend Snippet: LRIG1 expression in SHG-44 cells transfected with pEGFP-C1-LRIG1. a LRIG1 expression in the transfected cells detected by fluorescence microscopy 24 h after transfection. b Western blot analysis of LRIG1 expression. The results were normalized to the untransfected group and presented as means ± SD ( N = 4). *** P

    Article Snippet: The eukaryotic expression vector pEGFP-C1 was purchased from Clontech (Palo Alto, CA, USA).

    Techniques: Expressing, Transfection, Fluorescence, Microscopy, Western Blot

    RT-PCR products of five recombinant plasmids. M: 100-6000 bp DNA Marker. (A) Lane 1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3,Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the primer of β-actin. (B) Lane 1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3, Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the primer of hIL-18 full length. Lane 21 is negative control. (C) Lane1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3, Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the original primers. (D) Control. Lane 1-4 and lane 5-8 showed RT-PCR results of BxPC-3 cells tranfected with pEGFP-C1 and untreated BxPC-3 cells cultivated at 12 h, 24 h, 36 h, 48 h respectively, using the primer of hIL-18 full length. Lane 9-12 and lane 13-16 showed RT-PCR results of BxPC-3 cells with the same treatment (according to Lane 1-4 and 5-8) using the primer of β-actin.

    Journal: International Journal of Medical Sciences

    Article Title: Intracellular Distributing and Interferon-? Secretion of Human Interleukin-18 in BxPC-3 Cells

    doi: 10.7150/ijms.6875

    Figure Lengend Snippet: RT-PCR products of five recombinant plasmids. M: 100-6000 bp DNA Marker. (A) Lane 1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3,Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the primer of β-actin. (B) Lane 1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3, Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the primer of hIL-18 full length. Lane 21 is negative control. (C) Lane1-20 showed the RT-PCR results of BxPC-3 cells tranfected with five recombinant plasmids (Mu0, Mu1, Mu2, Mu3, Mu4) at 12 h, 24 h, 36 h, 48 h respectively, using the original primers. (D) Control. Lane 1-4 and lane 5-8 showed RT-PCR results of BxPC-3 cells tranfected with pEGFP-C1 and untreated BxPC-3 cells cultivated at 12 h, 24 h, 36 h, 48 h respectively, using the primer of hIL-18 full length. Lane 9-12 and lane 13-16 showed RT-PCR results of BxPC-3 cells with the same treatment (according to Lane 1-4 and 5-8) using the primer of β-actin.

    Article Snippet: 2.1 Plasmids and cell culture Eukaryotic expression vector pEGFP-C1 was purchased from Clontech (Palo Alto, USA). pGEM-T-hIL-18 was conserved in our laboratory.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Recombinant, Marker, Negative Control

    Localization of enhanced green fluorescent protein (EGFP)-tagged hIL-18 recombinant plasmids inBxPC-3 cells. Confocal micrographs of transfected BxPC-3 cells expressing EGFP-tagged hIL-18 recombinant plasmid at 12 h post-transfection (A) 24 h post-transfection, (B) 36 h post-transfection, (C) 48 h post-transfection, (D) and composite photo (E) respectively . Images were taken in the plane where maximum fluorescence appeared. Transfected cells presented uniform fluorescence throughout the cytoplasm and the nucleus 12 h post-transfection, and the fluorescence intensity of transfected cells with pEGFP-C1 are durable(A-D), fluorescence intensity of other cells are decreasing. But from 24 h and 36 h post-transfection, the fluorescence of the hIL-18 Mu1 and Mu2 appeared targeted to the membranous region of the BxPC-3 cells. Magnification, 200×.

    Journal: International Journal of Medical Sciences

    Article Title: Intracellular Distributing and Interferon-? Secretion of Human Interleukin-18 in BxPC-3 Cells

    doi: 10.7150/ijms.6875

    Figure Lengend Snippet: Localization of enhanced green fluorescent protein (EGFP)-tagged hIL-18 recombinant plasmids inBxPC-3 cells. Confocal micrographs of transfected BxPC-3 cells expressing EGFP-tagged hIL-18 recombinant plasmid at 12 h post-transfection (A) 24 h post-transfection, (B) 36 h post-transfection, (C) 48 h post-transfection, (D) and composite photo (E) respectively . Images were taken in the plane where maximum fluorescence appeared. Transfected cells presented uniform fluorescence throughout the cytoplasm and the nucleus 12 h post-transfection, and the fluorescence intensity of transfected cells with pEGFP-C1 are durable(A-D), fluorescence intensity of other cells are decreasing. But from 24 h and 36 h post-transfection, the fluorescence of the hIL-18 Mu1 and Mu2 appeared targeted to the membranous region of the BxPC-3 cells. Magnification, 200×.

    Article Snippet: 2.1 Plasmids and cell culture Eukaryotic expression vector pEGFP-C1 was purchased from Clontech (Palo Alto, USA). pGEM-T-hIL-18 was conserved in our laboratory.

    Techniques: Recombinant, Transfection, Expressing, Plasmid Preparation, Fluorescence