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pmax e2f1 expression vector  (Addgene inc)


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    Addgene inc pmax e2f1 expression vector
    <t>E2F1</t> upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a <t>pMax-E2F1</t> expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.
    Pmax E2f1 Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e2f1+expression+vector/pMax-E2F1+(Plasmid+%2316007)/pmc06089107-40-1-7
    Average 93 stars, based on 18 article reviews
    pmax e2f1 expression vector - by Bioz Stars, 2026-10
    93/100 stars

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    1) Product Images from "E2F1 induces LSINCT5 transcriptional activity and promotes gastric cancer progression by affecting the epithelial-mesenchymal transition"

    Article Title: E2F1 induces LSINCT5 transcriptional activity and promotes gastric cancer progression by affecting the epithelial-mesenchymal transition

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S171652

    E2F1 upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a pMax-E2F1 expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.
    Figure Legend Snippet: E2F1 upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a pMax-E2F1 expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.

    Techniques Used: Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Cotransfection, Over Expression, Construct, Transfection, Activity Assay, Plasmid Preparation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control

    Primers used in this study
    Figure Legend Snippet: Primers used in this study

    Techniques Used: Real-time Polymerase Chain Reaction

    Related Articles

    Plasmid Preparation:

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector (EV) expression plasmid (pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene) was transiently transfected GTL-16 cells using LipofectamineTM LTX reagent (Invitrogen) and kept until 72 hr post transfection (unless stated otherwise). ..

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector expression plasmid [pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene] was transiently transfected with GTL-16 cells using Lipofectamine LTX reagent (Invitrogen) and kept until 72 hours post transfection (unless stated otherwise). ..

    Article Title: Essential gene knockdowns reveal genetic vulnerabilities and antibiotic sensitivities in Acinetobacter baumannii .
    Article Snippet: .. The Mobile-CRISPRi plasmid pJMP2748 is a variant of pJMP2754 (Addgene 160666) with the sgRNA promoter derived from pJMP2367 (Addgene 160076) and the dCas9 promoter region amplified from the A. baumannii suppressor strain gDNA with oJMP635 and oJMP636. ..

    Article Title: Sustained cancer‐relevant alternative RNA splicing events driven by PRMT5 in high‐risk neuroblastoma
    Article Snippet: RNA interference was carried out with 15 n m siRNA using the Oligofectamine transfection reagent (Invitrogen, Waltham, MA, USA), as per the manufacturer's instructions. siRNAs used are as follows: non‐targeting control, 5′‐AGCUGACCCUGAAGUUCUU‐3′, CPSF3 (human, ID: s28531, Life Technologies, Carlsbad, CA, USA), CPSF4 (human, ID: s21412, Life Technologies), HNRNPA3 (human, ID: s47927, Life Technologies), HNRNPM (human, ID: s144164, Life Technologies), SRSF1 (human, ID: s143028, Life Technologies), SRSF4 (human, ID: s12734, Life Technologies) and DIABLO (human, ID:s224426, Life Technologies). .. Stable GFP‐E2F1 expressing CHP‐134 cell lines were generated by transfecting a pMaX‐GFP vector containing a full‐length E2F1 insert (plasmid #16007, Addgene, Watertown, MA, USA) into CHP‐134 E2F1 Cr (CRISPR) cells and selecting with G418. ..

    Expressing:

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector (EV) expression plasmid (pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene) was transiently transfected GTL-16 cells using LipofectamineTM LTX reagent (Invitrogen) and kept until 72 hr post transfection (unless stated otherwise). ..

    Article Title: High-throughput optimization of antibody production in CHO cells by tuning heavy- and light-chain promoter strength
    Article Snippet: Escherichia coli Mach1 T1R chemically competent cells (C869601, Thermo Fisher Scientific, Waltham, MA, United States) 48 well LB agar plates (prepared in-house and supplemented with 60 μg/mL ampicillin) 2× YT medium: 16 g tryptone, 10 g yeast extract, and 5 g NaCl per liter of distilled water; pH adjusted to 7.0 and sterilized by autoclaving (121 °C, 20 min) SOC medium: prepared according to standard formulations (2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl 2 , 10 mM MgSO 4 , and 20 mM glucose) Ampicillin (60 μg/mL, final concentration in liquid culture) .. CHO-S cells (R80007, Life Technologies, Waltham, MA, United States) CD CHO medium (10743029, Gibco, Thermo Fisher Scientific, Waltham, MA, United States) L-Glutamine (25030081, Thermo Fisher Scientific, Waltham, MA, United States) Freestyle MAX Transfection Reagent (16447100, Thermo Fisher Scientific, Waltham, MA, United States) OptiPRO serum free medium (12309019, Thermo Fisher Scientific, Waltham, MA, United States) Transfection positive control expressing GFP (pMax-E2F1, 16007, Addgene, Watertown, MA, United States) Solution 18 (910-3018, ChemoMetec A/S, Allerød, Denmark) NC-Slide A8 (941-0002, ChemoMetec A/S, Allerød, Denmark) .. MicroAmp Fast 96-Well Reaction Plate (4346907, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) SimpliAmp Thermal Cycler (A24811, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) Filtered pipette tips (10 μL, 200 μL, and 1000 μL; Thermo Fisher Scientific, Waltham, MA, United States) 1.5 mL and 2.0 mL microcentrifuge tubes (Eppendorf SE, Hamburg, Germany) Benchtop microcentrifuge (Eppendorf SE, Hamburg, Germany) Eppendorf Centrifuge 5810 R (Eppendorf SE, Hamburg, Germany) Gel electrophoresis system (Thermo Fisher Scientific, Waltham, MA, United States) 125 mL Erlenmeyer flasks with vent caps (431143, Corning Inc., Corning, NY, United States) 96-half-deep-well microplate (CR1496c, Enzyscreen B.V., Haarlem, The Netherlands) Autoclaved low-evaporation sandwich Duetz covers (CR1296a, Enzyscreen B.V., Heemstede, The Netherlands) Octet RED96 System (Sartorius, Göttingen, Germany) Protein A Biosensors (18-5010, Sartorius, Göttingen, Germany) Black 96-Well Microplates, Non-Binding Surface (3694, Corning Inc., Corning, NY, United States) Shaking incubator (37 °C, 350 rpm) Eppendorf New Brunswick S41i CO 2 incubator for mammalian cell culture (37 °C, 5% CO 2 , 70% humidity, 325 rpm, 2.5 cm orbital diameter) Amersham Imager 600 (AI600, Cytiva, Marlborough, MA, United States) NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, United States) NucleoCounter NC-200 (ChemoMetec A/S, Allerød, Denmark) CytoFLEX flow cytometer (Beckman Coulter, United States) Sterile disposable inoculating loop (10 μL; Deltalab, Barcelona, Spain)

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector expression plasmid [pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene] was transiently transfected with GTL-16 cells using Lipofectamine LTX reagent (Invitrogen) and kept until 72 hours post transfection (unless stated otherwise). ..

    Article Title: Sustained cancer‐relevant alternative RNA splicing events driven by PRMT5 in high‐risk neuroblastoma
    Article Snippet: RNA interference was carried out with 15 n m siRNA using the Oligofectamine transfection reagent (Invitrogen, Waltham, MA, USA), as per the manufacturer's instructions. siRNAs used are as follows: non‐targeting control, 5′‐AGCUGACCCUGAAGUUCUU‐3′, CPSF3 (human, ID: s28531, Life Technologies, Carlsbad, CA, USA), CPSF4 (human, ID: s21412, Life Technologies), HNRNPA3 (human, ID: s47927, Life Technologies), HNRNPM (human, ID: s144164, Life Technologies), SRSF1 (human, ID: s143028, Life Technologies), SRSF4 (human, ID: s12734, Life Technologies) and DIABLO (human, ID:s224426, Life Technologies). .. Stable GFP‐E2F1 expressing CHP‐134 cell lines were generated by transfecting a pMaX‐GFP vector containing a full‐length E2F1 insert (plasmid #16007, Addgene, Watertown, MA, USA) into CHP‐134 E2F1 Cr (CRISPR) cells and selecting with G418. ..

    Transfection:

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector (EV) expression plasmid (pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene) was transiently transfected GTL-16 cells using LipofectamineTM LTX reagent (Invitrogen) and kept until 72 hr post transfection (unless stated otherwise). ..

    Article Title: High-throughput optimization of antibody production in CHO cells by tuning heavy- and light-chain promoter strength
    Article Snippet: Escherichia coli Mach1 T1R chemically competent cells (C869601, Thermo Fisher Scientific, Waltham, MA, United States) 48 well LB agar plates (prepared in-house and supplemented with 60 μg/mL ampicillin) 2× YT medium: 16 g tryptone, 10 g yeast extract, and 5 g NaCl per liter of distilled water; pH adjusted to 7.0 and sterilized by autoclaving (121 °C, 20 min) SOC medium: prepared according to standard formulations (2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl 2 , 10 mM MgSO 4 , and 20 mM glucose) Ampicillin (60 μg/mL, final concentration in liquid culture) .. CHO-S cells (R80007, Life Technologies, Waltham, MA, United States) CD CHO medium (10743029, Gibco, Thermo Fisher Scientific, Waltham, MA, United States) L-Glutamine (25030081, Thermo Fisher Scientific, Waltham, MA, United States) Freestyle MAX Transfection Reagent (16447100, Thermo Fisher Scientific, Waltham, MA, United States) OptiPRO serum free medium (12309019, Thermo Fisher Scientific, Waltham, MA, United States) Transfection positive control expressing GFP (pMax-E2F1, 16007, Addgene, Watertown, MA, United States) Solution 18 (910-3018, ChemoMetec A/S, Allerød, Denmark) NC-Slide A8 (941-0002, ChemoMetec A/S, Allerød, Denmark) .. MicroAmp Fast 96-Well Reaction Plate (4346907, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) SimpliAmp Thermal Cycler (A24811, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) Filtered pipette tips (10 μL, 200 μL, and 1000 μL; Thermo Fisher Scientific, Waltham, MA, United States) 1.5 mL and 2.0 mL microcentrifuge tubes (Eppendorf SE, Hamburg, Germany) Benchtop microcentrifuge (Eppendorf SE, Hamburg, Germany) Eppendorf Centrifuge 5810 R (Eppendorf SE, Hamburg, Germany) Gel electrophoresis system (Thermo Fisher Scientific, Waltham, MA, United States) 125 mL Erlenmeyer flasks with vent caps (431143, Corning Inc., Corning, NY, United States) 96-half-deep-well microplate (CR1496c, Enzyscreen B.V., Haarlem, The Netherlands) Autoclaved low-evaporation sandwich Duetz covers (CR1296a, Enzyscreen B.V., Heemstede, The Netherlands) Octet RED96 System (Sartorius, Göttingen, Germany) Protein A Biosensors (18-5010, Sartorius, Göttingen, Germany) Black 96-Well Microplates, Non-Binding Surface (3694, Corning Inc., Corning, NY, United States) Shaking incubator (37 °C, 350 rpm) Eppendorf New Brunswick S41i CO 2 incubator for mammalian cell culture (37 °C, 5% CO 2 , 70% humidity, 325 rpm, 2.5 cm orbital diameter) Amersham Imager 600 (AI600, Cytiva, Marlborough, MA, United States) NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, United States) NucleoCounter NC-200 (ChemoMetec A/S, Allerød, Denmark) CytoFLEX flow cytometer (Beckman Coulter, United States) Sterile disposable inoculating loop (10 μL; Deltalab, Barcelona, Spain)

    Article Title: E2F1-Associated Purine Synthesis Pathway Is a Major Component of the MET-DNA Damage Response Network
    Article Snippet: .. An E2F1 or empty vector expression plasmid [pMax–E2F1 (RRID:Addgene_16007) or pMax-EV; Addgene] was transiently transfected with GTL-16 cells using Lipofectamine LTX reagent (Invitrogen) and kept until 72 hours post transfection (unless stated otherwise). ..

    Positive Control:

    Article Title: High-throughput optimization of antibody production in CHO cells by tuning heavy- and light-chain promoter strength
    Article Snippet: Escherichia coli Mach1 T1R chemically competent cells (C869601, Thermo Fisher Scientific, Waltham, MA, United States) 48 well LB agar plates (prepared in-house and supplemented with 60 μg/mL ampicillin) 2× YT medium: 16 g tryptone, 10 g yeast extract, and 5 g NaCl per liter of distilled water; pH adjusted to 7.0 and sterilized by autoclaving (121 °C, 20 min) SOC medium: prepared according to standard formulations (2% tryptone, 0.5% yeast extract, 10 mM NaCl, 2.5 mM KCl, 10 mM MgCl 2 , 10 mM MgSO 4 , and 20 mM glucose) Ampicillin (60 μg/mL, final concentration in liquid culture) .. CHO-S cells (R80007, Life Technologies, Waltham, MA, United States) CD CHO medium (10743029, Gibco, Thermo Fisher Scientific, Waltham, MA, United States) L-Glutamine (25030081, Thermo Fisher Scientific, Waltham, MA, United States) Freestyle MAX Transfection Reagent (16447100, Thermo Fisher Scientific, Waltham, MA, United States) OptiPRO serum free medium (12309019, Thermo Fisher Scientific, Waltham, MA, United States) Transfection positive control expressing GFP (pMax-E2F1, 16007, Addgene, Watertown, MA, United States) Solution 18 (910-3018, ChemoMetec A/S, Allerød, Denmark) NC-Slide A8 (941-0002, ChemoMetec A/S, Allerød, Denmark) .. MicroAmp Fast 96-Well Reaction Plate (4346907, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) SimpliAmp Thermal Cycler (A24811, Applied Biosystems, Thermo Fisher Scientific, Waltham, MA, United States) Filtered pipette tips (10 μL, 200 μL, and 1000 μL; Thermo Fisher Scientific, Waltham, MA, United States) 1.5 mL and 2.0 mL microcentrifuge tubes (Eppendorf SE, Hamburg, Germany) Benchtop microcentrifuge (Eppendorf SE, Hamburg, Germany) Eppendorf Centrifuge 5810 R (Eppendorf SE, Hamburg, Germany) Gel electrophoresis system (Thermo Fisher Scientific, Waltham, MA, United States) 125 mL Erlenmeyer flasks with vent caps (431143, Corning Inc., Corning, NY, United States) 96-half-deep-well microplate (CR1496c, Enzyscreen B.V., Haarlem, The Netherlands) Autoclaved low-evaporation sandwich Duetz covers (CR1296a, Enzyscreen B.V., Heemstede, The Netherlands) Octet RED96 System (Sartorius, Göttingen, Germany) Protein A Biosensors (18-5010, Sartorius, Göttingen, Germany) Black 96-Well Microplates, Non-Binding Surface (3694, Corning Inc., Corning, NY, United States) Shaking incubator (37 °C, 350 rpm) Eppendorf New Brunswick S41i CO 2 incubator for mammalian cell culture (37 °C, 5% CO 2 , 70% humidity, 325 rpm, 2.5 cm orbital diameter) Amersham Imager 600 (AI600, Cytiva, Marlborough, MA, United States) NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, United States) NucleoCounter NC-200 (ChemoMetec A/S, Allerød, Denmark) CytoFLEX flow cytometer (Beckman Coulter, United States) Sterile disposable inoculating loop (10 μL; Deltalab, Barcelona, Spain)

    Variant Assay:

    Article Title: Essential gene knockdowns reveal genetic vulnerabilities and antibiotic sensitivities in Acinetobacter baumannii .
    Article Snippet: .. The Mobile-CRISPRi plasmid pJMP2748 is a variant of pJMP2754 (Addgene 160666) with the sgRNA promoter derived from pJMP2367 (Addgene 160076) and the dCas9 promoter region amplified from the A. baumannii suppressor strain gDNA with oJMP635 and oJMP636. ..

    Derivative Assay:

    Article Title: Essential gene knockdowns reveal genetic vulnerabilities and antibiotic sensitivities in Acinetobacter baumannii .
    Article Snippet: .. The Mobile-CRISPRi plasmid pJMP2748 is a variant of pJMP2754 (Addgene 160666) with the sgRNA promoter derived from pJMP2367 (Addgene 160076) and the dCas9 promoter region amplified from the A. baumannii suppressor strain gDNA with oJMP635 and oJMP636. ..

    Amplification:

    Article Title: Essential gene knockdowns reveal genetic vulnerabilities and antibiotic sensitivities in Acinetobacter baumannii .
    Article Snippet: .. The Mobile-CRISPRi plasmid pJMP2748 is a variant of pJMP2754 (Addgene 160666) with the sgRNA promoter derived from pJMP2367 (Addgene 160076) and the dCas9 promoter region amplified from the A. baumannii suppressor strain gDNA with oJMP635 and oJMP636. ..

    Generated:

    Article Title: Sustained cancer‐relevant alternative RNA splicing events driven by PRMT5 in high‐risk neuroblastoma
    Article Snippet: RNA interference was carried out with 15 n m siRNA using the Oligofectamine transfection reagent (Invitrogen, Waltham, MA, USA), as per the manufacturer's instructions. siRNAs used are as follows: non‐targeting control, 5′‐AGCUGACCCUGAAGUUCUU‐3′, CPSF3 (human, ID: s28531, Life Technologies, Carlsbad, CA, USA), CPSF4 (human, ID: s21412, Life Technologies), HNRNPA3 (human, ID: s47927, Life Technologies), HNRNPM (human, ID: s144164, Life Technologies), SRSF1 (human, ID: s143028, Life Technologies), SRSF4 (human, ID: s12734, Life Technologies) and DIABLO (human, ID:s224426, Life Technologies). .. Stable GFP‐E2F1 expressing CHP‐134 cell lines were generated by transfecting a pMaX‐GFP vector containing a full‐length E2F1 insert (plasmid #16007, Addgene, Watertown, MA, USA) into CHP‐134 E2F1 Cr (CRISPR) cells and selecting with G418. ..

    CRISPR:

    Article Title: Sustained cancer‐relevant alternative RNA splicing events driven by PRMT5 in high‐risk neuroblastoma
    Article Snippet: RNA interference was carried out with 15 n m siRNA using the Oligofectamine transfection reagent (Invitrogen, Waltham, MA, USA), as per the manufacturer's instructions. siRNAs used are as follows: non‐targeting control, 5′‐AGCUGACCCUGAAGUUCUU‐3′, CPSF3 (human, ID: s28531, Life Technologies, Carlsbad, CA, USA), CPSF4 (human, ID: s21412, Life Technologies), HNRNPA3 (human, ID: s47927, Life Technologies), HNRNPM (human, ID: s144164, Life Technologies), SRSF1 (human, ID: s143028, Life Technologies), SRSF4 (human, ID: s12734, Life Technologies) and DIABLO (human, ID:s224426, Life Technologies). .. Stable GFP‐E2F1 expressing CHP‐134 cell lines were generated by transfecting a pMaX‐GFP vector containing a full‐length E2F1 insert (plasmid #16007, Addgene, Watertown, MA, USA) into CHP‐134 E2F1 Cr (CRISPR) cells and selecting with G418. ..



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    Addgene inc e2f1 expression vector pcmvha e2f1
    <t>E2F1</t> upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a <t>pMax-E2F1</t> expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.
    E2f1 Expression Vector Pcmvha E2f1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e2f1+expression+vector/pCMVHA+E2F1+(Plasmid+%2324225)/pm32354165-77-11-22
    Average 94 stars, based on 1 article reviews
    e2f1 expression vector pcmvha e2f1 - by Bioz Stars, 2026-10
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    e2f1 expression vector pcmv-e2f1  (Thermo Fisher)
    90
    Thermo Fisher e2f1 expression vector pcmv-e2f1
    Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes ( AHR , THBS1 , DDIT3 , and MKNK2 ) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes ( <t>E2F1</t> , PCNA , CCND1 , CCNE1 MCM2 , MCM4 , MCM5 , MCM7 , and KIAA0101 ) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1 , CCND1 , CCNE1 , MCM2 -7 and GAPDH . The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.
    E2f1 Expression Vector Pcmv E2f1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e2f1+expression+vector/pmc07509406-81-1-11
    Average 90 stars, based on 1 article reviews
    e2f1 expression vector pcmv-e2f1 - by Bioz Stars, 2026-10
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    e2f1 expression vector e2f1 ha  (Addgene inc)
    94
    Addgene inc e2f1 expression vector e2f1 ha
    Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes ( AHR , THBS1 , DDIT3 , and MKNK2 ) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes ( <t>E2F1</t> , PCNA , CCND1 , CCNE1 MCM2 , MCM4 , MCM5 , MCM7 , and KIAA0101 ) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1 , CCND1 , CCNE1 , MCM2 -7 and GAPDH . The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.
    E2f1 Expression Vector E2f1 Ha, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e2f1+expression+vector/pCMVHA+E2F1+(Plasmid+%2324225)/ppr0125839-93-9-17
    Average 94 stars, based on 1 article reviews
    e2f1 expression vector e2f1 ha - by Bioz Stars, 2026-10
    94/100 stars
    		  Buy from Supplier
    e2f1 expression vector  (Addgene inc)
    96
    Addgene inc e2f1 expression vector
    Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes ( AHR , THBS1 , DDIT3 , and MKNK2 ) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes ( <t>E2F1</t> , PCNA , CCND1 , CCNE1 MCM2 , MCM4 , MCM5 , MCM7 , and KIAA0101 ) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1 , CCND1 , CCNE1 , MCM2 -7 and GAPDH . The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.
    E2f1 Expression Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/e2f1+expression+vector/pcDNA3%2E1(%2B)+Laccase2+MCS+Exon+Vector+(Plasmid+%2369893)/pmc07086368__thnov10p3952s1-19-9-13
    Average 96 stars, based on 1 article reviews
    e2f1 expression vector - by Bioz Stars, 2026-10
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    Image Search Results


    E2F1 upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a pMax-E2F1 expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.

    Journal: Cancer Management and Research

    Article Title: E2F1 induces LSINCT5 transcriptional activity and promotes gastric cancer progression by affecting the epithelial-mesenchymal transition

    doi: 10.2147/CMAR.S171652

    Figure Lengend Snippet: E2F1 upregulates LSINCT5 expression in GC cells. Notes: qRT-PCR analysis of the E2F1 expression levels in GC tissues and paired ANTs ( A ) and in metastatic and nonmetastatic GC tissues ( B ). ( C ) qRT-PCR analysis of E2F1 expression in a series of human GC cell lines and a normal gastric epithelial cell line (GES-1). ( D ) ChIP assays using E2F1 antibody demonstrated endogenous E2F1 binding to the LSINCT5 gene promoter, and ectopic expression or siRNA knockdown increased or reduced E2F1 enrichment at the LSINCT5 promoter, respectively. ( E ) A dual-luciferase reporter assay was performed through cotransfection of the LSINCT5 promoter fragment (pGL3-LSINCT5) and an E2F1-overexpression construct. ( F ) We examined the LSINCT5 core promoter region for transcription factor binding sites, and identified four tandem putative E2F1-binding sites. Reporter assays were conducted in cells transfected with various LSINCT5 promoter constructs in which different E2F1-binding elements were deleted (WT, del). Luciferase activity is presented relative to that of the pGL3 vector (a promoter-less vector). ( G ) qRT-PCR analysis of the LSINCT5 expression levels following the transfection of MGC803 and BGC823 cells with a pMax-E2F1 expression vector and siRNA-E2F1, respectively. ( H ) Analysis of the relationship between LSINCT5 and E2F1 mRNA levels (ΔCt value) in GC tissues. Bars: SD; * P <0.05 and ** P <0.01. Abbreviations: ANTs, adjacent normal tissues; Ts, tumor tissues; ChIP, chromatin immunoprecipitation; del, deletion; GC, gastric cancer; qRT-PCR, quantitative real-time polymerase chain reaction; WT, wild type; NC, negative control.

    Article Snippet: The pMax-E2F1 expression vector was purchased from Addgene (plasmid #16007) (Cambridge, MA, USA), and siRNAs specific for E2F1 (#AM16708) and LSINCT5 (#4392420) were chemically synthesized (Invitrogen, Carlsbad, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Cotransfection, Over Expression, Construct, Transfection, Activity Assay, Plasmid Preparation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control

    Primers used in this study

    Journal: Cancer Management and Research

    Article Title: E2F1 induces LSINCT5 transcriptional activity and promotes gastric cancer progression by affecting the epithelial-mesenchymal transition

    doi: 10.2147/CMAR.S171652

    Figure Lengend Snippet: Primers used in this study

    Article Snippet: The pMax-E2F1 expression vector was purchased from Addgene (plasmid #16007) (Cambridge, MA, USA), and siRNAs specific for E2F1 (#AM16708) and LSINCT5 (#4392420) were chemically synthesized (Invitrogen, Carlsbad, CA, USA).

    Techniques: Real-time Polymerase Chain Reaction

    Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes ( AHR , THBS1 , DDIT3 , and MKNK2 ) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes ( E2F1 , PCNA , CCND1 , CCNE1 MCM2 , MCM4 , MCM5 , MCM7 , and KIAA0101 ) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1 , CCND1 , CCNE1 , MCM2 -7 and GAPDH . The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.

    Journal: Frontiers in Oncology

    Article Title: RPL21 siRNA Blocks Proliferation in Pancreatic Cancer Cells by Inhibiting DNA Replication and Inducing G1 Arrest and Apoptosis

    doi: 10.3389/fonc.2020.01730

    Figure Lengend Snippet: Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes ( AHR , THBS1 , DDIT3 , and MKNK2 ) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes ( E2F1 , PCNA , CCND1 , CCNE1 MCM2 , MCM4 , MCM5 , MCM7 , and KIAA0101 ) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1 , CCND1 , CCNE1 , MCM2 -7 and GAPDH . The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.

    Article Snippet: The E2F1 expression vector pCMV-E2F1 was constructed using the pcDNA3.1 plasmid (Invitrogen, Carlsbad, CA, United States).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Sequencing, Western Blot, Negative Control

    RPL21 regulates the cell cycle and DNA replication via E2F1 in PANC-1 and BxPC-3 cells. (A,C) The luciferase reporter vectors ( CCND1 , CCNE1 and MCM2 - 7 ) were constructed using pGL-3 vectors. 1 μg reporter vector was co-transfected with 1 μg pCMV-E2F1 vector or 1 μg empty pcDNA3.1 vector (control) for each well (6-well plates) containing 6 × 10 5 PANC-1 and BxPC-3 cells. Luciferase activity was measured 24 h after the transfection. (B,D) The PANC-1 and BxPC-3 cells were transfected with siRNAs with lipofectamine 2000 reagent. The control, NC, siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. The cells after transfection were seeded in 6-well plates at 6 × 10 5 cells/well, and 2 g E2F1 luciferase reporters (E2F1-promoter) were transfected using lipofectamine 2000 reagent. Luciferase activity was measured 24 h after the transfection. Each bar represents the mean ± SD of triplicate analysis. * indicates P < 0.05 compared to the control as determined by the Student’s t -test.

    Journal: Frontiers in Oncology

    Article Title: RPL21 siRNA Blocks Proliferation in Pancreatic Cancer Cells by Inhibiting DNA Replication and Inducing G1 Arrest and Apoptosis

    doi: 10.3389/fonc.2020.01730

    Figure Lengend Snippet: RPL21 regulates the cell cycle and DNA replication via E2F1 in PANC-1 and BxPC-3 cells. (A,C) The luciferase reporter vectors ( CCND1 , CCNE1 and MCM2 - 7 ) were constructed using pGL-3 vectors. 1 μg reporter vector was co-transfected with 1 μg pCMV-E2F1 vector or 1 μg empty pcDNA3.1 vector (control) for each well (6-well plates) containing 6 × 10 5 PANC-1 and BxPC-3 cells. Luciferase activity was measured 24 h after the transfection. (B,D) The PANC-1 and BxPC-3 cells were transfected with siRNAs with lipofectamine 2000 reagent. The control, NC, siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. The cells after transfection were seeded in 6-well plates at 6 × 10 5 cells/well, and 2 g E2F1 luciferase reporters (E2F1-promoter) were transfected using lipofectamine 2000 reagent. Luciferase activity was measured 24 h after the transfection. Each bar represents the mean ± SD of triplicate analysis. * indicates P < 0.05 compared to the control as determined by the Student’s t -test.

    Article Snippet: The E2F1 expression vector pCMV-E2F1 was constructed using the pcDNA3.1 plasmid (Invitrogen, Carlsbad, CA, United States).

    Techniques: Luciferase, Construct, Plasmid Preparation, Transfection, Activity Assay

    Model of E2F1 -mediated pancreatic cancer cell proliferation.

    Journal: Frontiers in Oncology

    Article Title: RPL21 siRNA Blocks Proliferation in Pancreatic Cancer Cells by Inhibiting DNA Replication and Inducing G1 Arrest and Apoptosis

    doi: 10.3389/fonc.2020.01730

    Figure Lengend Snippet: Model of E2F1 -mediated pancreatic cancer cell proliferation.

    Article Snippet: The E2F1 expression vector pCMV-E2F1 was constructed using the pcDNA3.1 plasmid (Invitrogen, Carlsbad, CA, United States).

    Techniques: