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pegfpc1 vector  (TaKaRa)


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    Structured Review

    TaKaRa pegfpc1 vector
    HEK293T cells were transfected with <t>pEGFPC1</t> vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.
    Pegfpc1 Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 619 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pegfpc1+vector/pmCherry+Vector/bio_rxiv__2023__07__24__550336-71-15-17
    Average 96 stars, based on 619 article reviews
    pegfpc1 vector - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells"

    Article Title: Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells

    Journal: bioRxiv

    doi: 10.1101/2023.07.24.550336

    HEK293T cells were transfected with pEGFPC1 vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.
    Figure Legend Snippet: HEK293T cells were transfected with pEGFPC1 vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.

    Techniques Used: Transfection, Plasmid Preparation, Immunostaining, Expressing, Western Blot

    HEK293T cells were infected with CHIKV at 0.1 MOI. Scribble levels were analyzed at 24 and 48 hpi using western blot and quantitated using densitometry (A). Twenty-four hours post-infection, the cells were fixed, and immunostaining was performed for Scribble and CHIKV capsid and E2 proteins. Inset shows zoomed images of an E2 expressing cell. Arrowheads indicate Scribble puncta (B). Effect of TF on Scribble expression. HEK293T cells were transfected with pEGFPC1-TF and its mutants. Only vector and pEGFPC1-6K were also transfected, and Scribble levels were analyzed 48 h post-transfection using western blot and quantitated using densitometry (C). Electron microscopy (EM) imaging of CHIKV-infected HEK293T cells. HEK293T cells were infected with CHIKV at 1.0 MOI and Scribble was visualized using a gold-labeled secondary antibody (D-i). EM images of an infected cell at higher magnification (D-ii). Immunostaining for ubiquitin and Scribble in CHIKV infected HEK293T cells (E). Data represents mean ± SEM of two biological replicates.
    Figure Legend Snippet: HEK293T cells were infected with CHIKV at 0.1 MOI. Scribble levels were analyzed at 24 and 48 hpi using western blot and quantitated using densitometry (A). Twenty-four hours post-infection, the cells were fixed, and immunostaining was performed for Scribble and CHIKV capsid and E2 proteins. Inset shows zoomed images of an E2 expressing cell. Arrowheads indicate Scribble puncta (B). Effect of TF on Scribble expression. HEK293T cells were transfected with pEGFPC1-TF and its mutants. Only vector and pEGFPC1-6K were also transfected, and Scribble levels were analyzed 48 h post-transfection using western blot and quantitated using densitometry (C). Electron microscopy (EM) imaging of CHIKV-infected HEK293T cells. HEK293T cells were infected with CHIKV at 1.0 MOI and Scribble was visualized using a gold-labeled secondary antibody (D-i). EM images of an infected cell at higher magnification (D-ii). Immunostaining for ubiquitin and Scribble in CHIKV infected HEK293T cells (E). Data represents mean ± SEM of two biological replicates.

    Techniques Used: Infection, Western Blot, Immunostaining, Expressing, Transfection, Plasmid Preparation, Electron Microscopy, Imaging, Labeling

    Related Articles

    Clone Assay:

    Article Title: Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells
    Article Snippet: .. The cDNA sequences of CHIKV 6K and TF (Supplementary Figure S1) were cloned in the pEGFPC1 vector (Clontech) to generate fusion proteins tagged with the Enhanced Green Fluorescent Protein (EGFP) at the N-terminus. ..

    Article Title: Importin α as a molecular marker for investigating the microenvironment of micronuclei
    Article Snippet: The fluorescence images were observed using a confocal microscope (Leica TCS SP8 II; Leica Microsystems, Wetzlar, Germany). .. Human importin α1 (KPNA2) cDNA subcloned into the pGEX6P2 plasmid was gifted from Akira Tsujii (NIBIOHN) and then cloned into pEGFPC1 vector (Clontech, Mountain View, CA, USA). pEGFPC1-mouse importin α2 (KPNA2) was constructed as described previously ( ). ..

    Article Title: C-terminal cysteines of HRas control Erk signaling and 15-deoxy-Δ 12,14 -prostaglandin J 2 (15d-PGJ 2 ) mediated inhibition of myoblast differentiation
    Article Snippet: .. Unmutated and cysteine mutants of HRas WT [HRas WT, HRas-C181S, and HRas-C184S] and HRas V12 [HRas V12, HRas V12-C181S, HRas V12-C184S] were cloned in the pEGFPC1 vector (Clontech) by restriction digestion-ligation method. .. Constructs of wild-type HRas were PCR amplified from a previously available HRas construct in the lab with construct-specific primers using Phusion High Fidelity DNA Polymerase (Thermo Scientific) followed by restriction digestion of HRas constructs and the pEGFPC1 vector using Xho1/EcoR1 enzymes (New England Biolabs Inc.).

    Article Title: Electron tomography reveals changes in spatial distribution of UBTF1 and UBTF2 isoforms within nucleolar components during rRNA synthesis inhibition.
    Article Snippet: .. After a BspEI–BamHI digest, the PCR products were cloned into the Age I and BamH I sites of pEGFPC1 vector (Clontech Laboratories, USA). ..

    Article Title: The cytosolic N-terminal region of heterologously-expressed transmembrane channel-like protein 1 (TMC1) can be cleaved in HEK293 cells
    Article Snippet: We used mTmc1ex1 cDNA which was cloned from cochleas of male C57BL/6N mice in pcDNA3.1(+) vector (Takara Bio, Otsu, Japan), mammalian expression vectors, as reported previously [ ]. .. Mouse Tmc2 cDNA was amplified from a cDNA library of dorsal root ganglia of a C57BL/6J male mouse and cloned into pEGFPC1 vector (Takara Bio). .. The mutants of mTmc1 were made by site-directed mutagenesis using the PrimeSTAR Mutagenesis Basal kit (Takara Bio).

    Plasmid Preparation:

    Article Title: Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells
    Article Snippet: .. The cDNA sequences of CHIKV 6K and TF (Supplementary Figure S1) were cloned in the pEGFPC1 vector (Clontech) to generate fusion proteins tagged with the Enhanced Green Fluorescent Protein (EGFP) at the N-terminus. ..

    Article Title: Importin α as a molecular marker for investigating the microenvironment of micronuclei
    Article Snippet: The fluorescence images were observed using a confocal microscope (Leica TCS SP8 II; Leica Microsystems, Wetzlar, Germany). .. Human importin α1 (KPNA2) cDNA subcloned into the pGEX6P2 plasmid was gifted from Akira Tsujii (NIBIOHN) and then cloned into pEGFPC1 vector (Clontech, Mountain View, CA, USA). pEGFPC1-mouse importin α2 (KPNA2) was constructed as described previously ( ). ..

    Article Title: C-terminal cysteines of HRas control Erk signaling and 15-deoxy-Δ 12,14 -prostaglandin J 2 (15d-PGJ 2 ) mediated inhibition of myoblast differentiation
    Article Snippet: .. Unmutated and cysteine mutants of HRas WT [HRas WT, HRas-C181S, and HRas-C184S] and HRas V12 [HRas V12, HRas V12-C181S, HRas V12-C184S] were cloned in the pEGFPC1 vector (Clontech) by restriction digestion-ligation method. .. Constructs of wild-type HRas were PCR amplified from a previously available HRas construct in the lab with construct-specific primers using Phusion High Fidelity DNA Polymerase (Thermo Scientific) followed by restriction digestion of HRas constructs and the pEGFPC1 vector using Xho1/EcoR1 enzymes (New England Biolabs Inc.).

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Article Title: Electron tomography reveals changes in spatial distribution of UBTF1 and UBTF2 isoforms within nucleolar components during rRNA synthesis inhibition.
    Article Snippet: .. After a BspEI–BamHI digest, the PCR products were cloned into the Age I and BamH I sites of pEGFPC1 vector (Clontech Laboratories, USA). ..

    Article Title: The cytosolic N-terminal region of heterologously-expressed transmembrane channel-like protein 1 (TMC1) can be cleaved in HEK293 cells
    Article Snippet: We used mTmc1ex1 cDNA which was cloned from cochleas of male C57BL/6N mice in pcDNA3.1(+) vector (Takara Bio, Otsu, Japan), mammalian expression vectors, as reported previously [ ]. .. Mouse Tmc2 cDNA was amplified from a cDNA library of dorsal root ganglia of a C57BL/6J male mouse and cloned into pEGFPC1 vector (Takara Bio). .. The mutants of mTmc1 were made by site-directed mutagenesis using the PrimeSTAR Mutagenesis Basal kit (Takara Bio).

    Article Title: ZIP kinase phosphorylated and activated by Rho kinase/ROCK contributes to cytokinesis in mammalian cultured cells.
    Article Snippet: Cytokinesis of animal cells requires contraction of a contractile ring, composed of actin filaments and myosin II filaments.. Phosphorylation of myosin II regulatory light chain (MRLC) promotes contraction of the actomyosin ring by activating myosin II motor activity.. Both Rho-associated coiled-coil kinase (Rho kinase/ROCK) and Zipper-interacting protein kinase (ZIP kinase/ZIPK) have been reported to phosphorylate MRLC at the contractile ring.

    Construct:

    Article Title: Importin α as a molecular marker for investigating the microenvironment of micronuclei
    Article Snippet: The fluorescence images were observed using a confocal microscope (Leica TCS SP8 II; Leica Microsystems, Wetzlar, Germany). .. Human importin α1 (KPNA2) cDNA subcloned into the pGEX6P2 plasmid was gifted from Akira Tsujii (NIBIOHN) and then cloned into pEGFPC1 vector (Clontech, Mountain View, CA, USA). pEGFPC1-mouse importin α2 (KPNA2) was constructed as described previously ( ). ..

    Modification:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Mutagenesis:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Sequencing:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    FLAG-tag:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Transfection:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Concentration Assay:

    Article Title: Lateral access mechanism of LPA receptor probed by molecular dynamics simulation
    Article Snippet: .. A modified pEGFPC1 vector (Clontech) encoding the wild-type or a mutant LPA 6 , consisting of a haemagglutinin signal sequence, FLAG epitope tag and zebrafish LPA 6 (residues 1–312) [ ], was transfected together with a pCAGGS plasmid encoding alkaline phosphatase (AP)-tagged TGFα (AP-TGFα; human codon-optimized) into HEK293FT cells (seeded 1-day before the transfection in 4 ml per 6-cm culture dish at a cell concentration of 2 x 10 cells/ml) by using a polyethylenimine (PEI) transfection reagent (400 ng LPA 6 plasmid, 1 μg AP-TGFα plasmid, and 8 μl of 1 mg/ml PEI solution per 6-cm culture dish). ..

    Polymerase Chain Reaction:

    Article Title: Electron tomography reveals changes in spatial distribution of UBTF1 and UBTF2 isoforms within nucleolar components during rRNA synthesis inhibition.
    Article Snippet: .. After a BspEI–BamHI digest, the PCR products were cloned into the Age I and BamH I sites of pEGFPC1 vector (Clontech Laboratories, USA). ..

    Amplification:

    Article Title: The cytosolic N-terminal region of heterologously-expressed transmembrane channel-like protein 1 (TMC1) can be cleaved in HEK293 cells
    Article Snippet: We used mTmc1ex1 cDNA which was cloned from cochleas of male C57BL/6N mice in pcDNA3.1(+) vector (Takara Bio, Otsu, Japan), mammalian expression vectors, as reported previously [ ]. .. Mouse Tmc2 cDNA was amplified from a cDNA library of dorsal root ganglia of a C57BL/6J male mouse and cloned into pEGFPC1 vector (Takara Bio). .. The mutants of mTmc1 were made by site-directed mutagenesis using the PrimeSTAR Mutagenesis Basal kit (Takara Bio).

    cDNA Library Assay:

    Article Title: The cytosolic N-terminal region of heterologously-expressed transmembrane channel-like protein 1 (TMC1) can be cleaved in HEK293 cells
    Article Snippet: We used mTmc1ex1 cDNA which was cloned from cochleas of male C57BL/6N mice in pcDNA3.1(+) vector (Takara Bio, Otsu, Japan), mammalian expression vectors, as reported previously [ ]. .. Mouse Tmc2 cDNA was amplified from a cDNA library of dorsal root ganglia of a C57BL/6J male mouse and cloned into pEGFPC1 vector (Takara Bio). .. The mutants of mTmc1 were made by site-directed mutagenesis using the PrimeSTAR Mutagenesis Basal kit (Takara Bio).

    other:

    Article Title: FSP27 Promotes Lipid Droplet Clustering and Then Fusion to Regulate Triglyceride Accumulation
    Article Snippet: COS-7 cells were purchased from ATCC. pEGFPC1 vector was purchased from Clontech.

    Generated:

    Article Title: ZIP kinase phosphorylated and activated by Rho kinase/ROCK contributes to cytokinesis in mammalian cultured cells.
    Article Snippet: Cytokinesis of animal cells requires contraction of a contractile ring, composed of actin filaments and myosin II filaments.. Phosphorylation of myosin II regulatory light chain (MRLC) promotes contraction of the actomyosin ring by activating myosin II motor activity.. Both Rho-associated coiled-coil kinase (Rho kinase/ROCK) and Zipper-interacting protein kinase (ZIP kinase/ZIPK) have been reported to phosphorylate MRLC at the contractile ring.



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    HEK293T cells were transfected with <t>pEGFPC1</t> vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.
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    Image Search Results


    HEK293T cells were transfected with pEGFPC1 vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.

    Journal: bioRxiv

    Article Title: Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells

    doi: 10.1101/2023.07.24.550336

    Figure Lengend Snippet: HEK293T cells were transfected with pEGFPC1 vector or pEGFPC1 plasmids carrying TF or 6K. Twenty-four hours post-transfection, cells were fixed, and immunostaining was performed for Scribble (red). Inset shows zoomed images of an EGFP-TF expressing cell. Arrowheads indicate Scribble puncta and co-localization with TF (A). Mutational analysis of TF and its effect on Scribble localization (B). Expression data of TF and TF mutants in HEK293T cells. Twenty-four hours post-transfection, cells were lysed, and 6K and TF expression were analyzed using western blot with anti-GFP antibody (C). L: protein ladder, C1: pEGFPC1 empty vector, TF Mut1: TF-HGAATV, TF Mut2: TF-AAAATV.

    Article Snippet: The cDNA sequences of CHIKV 6K and TF (Supplementary Figure S1) were cloned in the pEGFPC1 vector (Clontech) to generate fusion proteins tagged with the Enhanced Green Fluorescent Protein (EGFP) at the N-terminus.

    Techniques: Transfection, Plasmid Preparation, Immunostaining, Expressing, Western Blot

    HEK293T cells were infected with CHIKV at 0.1 MOI. Scribble levels were analyzed at 24 and 48 hpi using western blot and quantitated using densitometry (A). Twenty-four hours post-infection, the cells were fixed, and immunostaining was performed for Scribble and CHIKV capsid and E2 proteins. Inset shows zoomed images of an E2 expressing cell. Arrowheads indicate Scribble puncta (B). Effect of TF on Scribble expression. HEK293T cells were transfected with pEGFPC1-TF and its mutants. Only vector and pEGFPC1-6K were also transfected, and Scribble levels were analyzed 48 h post-transfection using western blot and quantitated using densitometry (C). Electron microscopy (EM) imaging of CHIKV-infected HEK293T cells. HEK293T cells were infected with CHIKV at 1.0 MOI and Scribble was visualized using a gold-labeled secondary antibody (D-i). EM images of an infected cell at higher magnification (D-ii). Immunostaining for ubiquitin and Scribble in CHIKV infected HEK293T cells (E). Data represents mean ± SEM of two biological replicates.

    Journal: bioRxiv

    Article Title: Downregulation of a cell polarity protein potentiates Chikungunya Virus infection in host cells

    doi: 10.1101/2023.07.24.550336

    Figure Lengend Snippet: HEK293T cells were infected with CHIKV at 0.1 MOI. Scribble levels were analyzed at 24 and 48 hpi using western blot and quantitated using densitometry (A). Twenty-four hours post-infection, the cells were fixed, and immunostaining was performed for Scribble and CHIKV capsid and E2 proteins. Inset shows zoomed images of an E2 expressing cell. Arrowheads indicate Scribble puncta (B). Effect of TF on Scribble expression. HEK293T cells were transfected with pEGFPC1-TF and its mutants. Only vector and pEGFPC1-6K were also transfected, and Scribble levels were analyzed 48 h post-transfection using western blot and quantitated using densitometry (C). Electron microscopy (EM) imaging of CHIKV-infected HEK293T cells. HEK293T cells were infected with CHIKV at 1.0 MOI and Scribble was visualized using a gold-labeled secondary antibody (D-i). EM images of an infected cell at higher magnification (D-ii). Immunostaining for ubiquitin and Scribble in CHIKV infected HEK293T cells (E). Data represents mean ± SEM of two biological replicates.

    Article Snippet: The cDNA sequences of CHIKV 6K and TF (Supplementary Figure S1) were cloned in the pEGFPC1 vector (Clontech) to generate fusion proteins tagged with the Enhanced Green Fluorescent Protein (EGFP) at the N-terminus.

    Techniques: Infection, Western Blot, Immunostaining, Expressing, Transfection, Plasmid Preparation, Electron Microscopy, Imaging, Labeling

    Journal: eLife

    Article Title: SAFB regulates hippocampal stem cell fate by targeting Drosha to destabilize Nfib mRNA

    doi: 10.7554/eLife.74940

    Figure Lengend Snippet:

    Article Snippet: Recombinant DNA reagent , pEGFPC1-6XHis- FLKSRP (plasmid) , , RRID: Addgene_23001 , pEGFPC1 expression vector.

    Techniques: Protease Inhibitor, Recombinant, Clone Assay, Bicinchoninic Acid Protein Assay, Mutagenesis, Transfection, Sequencing, esiRNA, Blocking Assay, Plasmid Preparation, Expressing, Software