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are-pgl2 reporter plasmid  (Promega)

 
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    Promega are-pgl2 reporter plasmid
    Are Pgl2 Reporter Plasmid, supplied by Promega, 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/pgl2+reporter+plasmid/pgl3+basic/pmc12145549-299-1-38
    Average 90 stars, based on 1 article reviews
    are-pgl2 reporter plasmid - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: Regulation of expression of apolipoprotein A-I by selenium status in human liver hepatoblastoma cells.
    Article Snippet: Background Cardiomyopathy is common to areas with low selenium (Se) intake and in patients receiving total parenteral nutrition.. Although controversial, a few studies have suggested a protective role for Se in coronary heart disease on the basis of modulation of high-density lipoproteins (HDL).. Aims of the study In this study, the role of Se as a positive regulator of expression of a key HDL, apolipoprotein A-I (apoA-I), has been evaluated in human hepatoblastoma (HepG2) cell culture model. We further examined if the transcription of apoA-I, driven by the nuclear hormone receptor, peroxisome-proliferator activated receptor, PPARa, was trans-repressed by the presence of the oxidative stress-responsive transcription factor, NF-jB.

    Article Title: Lactobacillus acidophilus Alleviates Platelet-Activating Factor-Induced Inflammatory Responses in Human Intestinal Epithelial Cells
    Article Snippet: The secretion of IL-8 in the spent media of control and treated cells was measured by DuoSet ELISA kit for human IL-8 (R&D Systems, Minneapolis, MN), according to the manufacturer's instructions as described previously by us . .. A 1310 bp fragment of the 5 ′-untranslated region (p-Bcl1310) of Bcl10 gene cloned earlier into the pGL2 reporter plasmid (Promega, Madison, WI) was transfected into Caco-2 cells were using Lipofectamine 2000 (Invitrogen). ..

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. Cloning of 5′-untranslated region of Bcl10 gene and measurement of promoter activity A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: The probiotic Lactobacillus plantarum counteracts TNF-?-induced downregulation of SMCT1 expression and function
    Article Snippet: .. A 2,015-bp fragment of the 5′-untranslated region of slc5a8 gene and several 5′ deletion fragments were cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template.

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).

    Plasmid Preparation:

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Article Title: A gene regulatory architecture that controls region-independent dynamics of oligodendrocyte differentiation.
    Article Snippet: Laboratory of Systems Tumor Immunology, Hautklinik, Universitätsklinikum Erlangen and Friedrich-Alexander-Universität ErlangenNürnberg, Erlangen, Germany Faculty of Mechanical Engineering, Specialty Division for Systems Biotechnology, Technische Universität München, Munich, Germany Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität ErlangenNürnberg, Germany Neuroregeneration, Department of Neurology, Medical Faculty, Heinrich-Heine University, Düsseldorf, Germany Correspondence Prof. Dr. Julio Vera, Laboratory of Systems Tumor Immunology, Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstr.. 14, D-91052 Erlangen, Germany.. Email: julio.vera-gonzalez@uk-erlangen.de Funding information Deutsche Forschungsgemeinschaft, Grant/ Award Numbers: GRK2162/1, Ku1934/2-1, We1326/12-1, Ve642/1-1, Re3661/1-1 Abstract Oligodendrocytes (OLs) facilitate information processing in the vertebrate central nervous system via axonal ensheathment.

    Article Title: Regulation of expression of apolipoprotein A-I by selenium status in human liver hepatoblastoma cells.
    Article Snippet: Background Cardiomyopathy is common to areas with low selenium (Se) intake and in patients receiving total parenteral nutrition.. Although controversial, a few studies have suggested a protective role for Se in coronary heart disease on the basis of modulation of high-density lipoproteins (HDL).. Aims of the study In this study, the role of Se as a positive regulator of expression of a key HDL, apolipoprotein A-I (apoA-I), has been evaluated in human hepatoblastoma (HepG2) cell culture model. We further examined if the transcription of apoA-I, driven by the nuclear hormone receptor, peroxisome-proliferator activated receptor, PPARa, was trans-repressed by the presence of the oxidative stress-responsive transcription factor, NF-jB.

    Article Title: Lactobacillus acidophilus Alleviates Platelet-Activating Factor-Induced Inflammatory Responses in Human Intestinal Epithelial Cells
    Article Snippet: The secretion of IL-8 in the spent media of control and treated cells was measured by DuoSet ELISA kit for human IL-8 (R&D Systems, Minneapolis, MN), according to the manufacturer's instructions as described previously by us . .. A 1310 bp fragment of the 5 ′-untranslated region (p-Bcl1310) of Bcl10 gene cloned earlier into the pGL2 reporter plasmid (Promega, Madison, WI) was transfected into Caco-2 cells were using Lipofectamine 2000 (Invitrogen). ..

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. Cloning of 5′-untranslated region of Bcl10 gene and measurement of promoter activity A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: The probiotic Lactobacillus plantarum counteracts TNF-?-induced downregulation of SMCT1 expression and function
    Article Snippet: .. A 2,015-bp fragment of the 5′-untranslated region of slc5a8 gene and several 5′ deletion fragments were cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template.

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).

    Luciferase:

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Article Title: Regulation of expression of apolipoprotein A-I by selenium status in human liver hepatoblastoma cells.
    Article Snippet: Background Cardiomyopathy is common to areas with low selenium (Se) intake and in patients receiving total parenteral nutrition.. Although controversial, a few studies have suggested a protective role for Se in coronary heart disease on the basis of modulation of high-density lipoproteins (HDL).. Aims of the study In this study, the role of Se as a positive regulator of expression of a key HDL, apolipoprotein A-I (apoA-I), has been evaluated in human hepatoblastoma (HepG2) cell culture model. We further examined if the transcription of apoA-I, driven by the nuclear hormone receptor, peroxisome-proliferator activated receptor, PPARa, was trans-repressed by the presence of the oxidative stress-responsive transcription factor, NF-jB.

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. Cloning of 5′-untranslated region of Bcl10 gene and measurement of promoter activity A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: The probiotic Lactobacillus plantarum counteracts TNF-?-induced downregulation of SMCT1 expression and function
    Article Snippet: .. A 2,015-bp fragment of the 5′-untranslated region of slc5a8 gene and several 5′ deletion fragments were cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template.

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).

    Construct:

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Activity Assay:

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. Cloning of 5′-untranslated region of Bcl10 gene and measurement of promoter activity A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).

    Subcloning:

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Polymerase Chain Reaction:

    Article Title: Decreased AP-1 activity and interleukin-11 expression by bone marrow stromal cells may be associated with impaired bone formation in aged mice.
    Article Snippet: .. Construction of luciferase reporter plasmids Chimeric luciferase reporter plasmids, pGL2–3.8k,pGL2– 1.7k, pGL2–460, and pGL2–280, were constructed to investigate the transcriptional activity of the promoter by subcloning into the pGL2 reporter plasmid (Promega) polymerase chain reaction (PCR)-amplified fragments of the mouse IL-11 gene promoter extending from nucleotides 3800 to 6 (pGL2– 3.8k), 1700 to 6 (pGL2–1.7k), 445 to 6 (pGL2–460), and 269 to 6 (pGL2–280). ..

    Article Title: A gene regulatory architecture that controls region-independent dynamics of oligodendrocyte differentiation.
    Article Snippet: Laboratory of Systems Tumor Immunology, Hautklinik, Universitätsklinikum Erlangen and Friedrich-Alexander-Universität ErlangenNürnberg, Erlangen, Germany Faculty of Mechanical Engineering, Specialty Division for Systems Biotechnology, Technische Universität München, Munich, Germany Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität ErlangenNürnberg, Germany Neuroregeneration, Department of Neurology, Medical Faculty, Heinrich-Heine University, Düsseldorf, Germany Correspondence Prof. Dr. Julio Vera, Laboratory of Systems Tumor Immunology, Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstr.. 14, D-91052 Erlangen, Germany.. Email: julio.vera-gonzalez@uk-erlangen.de Funding information Deutsche Forschungsgemeinschaft, Grant/ Award Numbers: GRK2162/1, Ku1934/2-1, We1326/12-1, Ve642/1-1, Re3661/1-1 Abstract Oligodendrocytes (OLs) facilitate information processing in the vertebrate central nervous system via axonal ensheathment.

    Generated:

    Article Title: A gene regulatory architecture that controls region-independent dynamics of oligodendrocyte differentiation.
    Article Snippet: Laboratory of Systems Tumor Immunology, Hautklinik, Universitätsklinikum Erlangen and Friedrich-Alexander-Universität ErlangenNürnberg, Erlangen, Germany Faculty of Mechanical Engineering, Specialty Division for Systems Biotechnology, Technische Universität München, Munich, Germany Institut für Biochemie, Emil-Fischer-Zentrum, Friedrich-Alexander-Universität ErlangenNürnberg, Germany Neuroregeneration, Department of Neurology, Medical Faculty, Heinrich-Heine University, Düsseldorf, Germany Correspondence Prof. Dr. Julio Vera, Laboratory of Systems Tumor Immunology, Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstr.. 14, D-91052 Erlangen, Germany.. Email: julio.vera-gonzalez@uk-erlangen.de Funding information Deutsche Forschungsgemeinschaft, Grant/ Award Numbers: GRK2162/1, Ku1934/2-1, We1326/12-1, Ve642/1-1, Re3661/1-1 Abstract Oligodendrocytes (OLs) facilitate information processing in the vertebrate central nervous system via axonal ensheathment.

    Transfection:

    Article Title: Lactobacillus acidophilus Alleviates Platelet-Activating Factor-Induced Inflammatory Responses in Human Intestinal Epithelial Cells
    Article Snippet: The secretion of IL-8 in the spent media of control and treated cells was measured by DuoSet ELISA kit for human IL-8 (R&D Systems, Minneapolis, MN), according to the manufacturer's instructions as described previously by us . .. A 1310 bp fragment of the 5 ′-untranslated region (p-Bcl1310) of Bcl10 gene cloned earlier into the pGL2 reporter plasmid (Promega, Madison, WI) was transfected into Caco-2 cells were using Lipofectamine 2000 (Invitrogen). ..

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).

    Cloning:

    Article Title: Platelet-activating factor-induced NF-? B activation and IL-8 production in intestinal epithelial cells are Bcl10 dependent
    Article Snippet: The sample values were normalized with total protein content (BCA Protein assay kit; Pierce) and expressed as picograms per milligram cellular protein. .. Cloning of 5′-untranslated region of Bcl10 gene and measurement of promoter activity A 1310 bp fragment of the 5′-untranslated region of Bcl10 gene was cloned into the pGL2 reporter plasmid (Promega, Madison, WI) between Xho I and HinD III sites upstream of the luciferase reporter gene. .. We used Elongase Amplification System (Invitrogen) and PCR method to clone this fragment using human genomic DNA as the template and the gene specific primer pairs shown in .

    Recombinant:

    Article Title: ARID1A-mutated cervical cancer depends on the activation of YAP signaling.
    Article Snippet: .. Human YAP1 gene promoter was cloned into pGL2 reporter plasmid (Promega, USA), and the recombinant plasmid pGL2-YAP1 promoter was transfected into C33A, CaSki, ME180 or HeLa cells. pRL-SV40 reporter plasmid with Renilla luciferase activity (Promega, USA) was used to normalize the transfection efficiency. .. After 24 h, the firefly and Renilla luciferase activities weremeasured by a Dual-Luciferase Reporter Assay Kit (Promega, USA).



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    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of <t>2×CLEAR</t> (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .
    Snail Pgl2 Snail Promoter Luciferase Reporter Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of <t>2×CLEAR</t> (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .
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    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of <t>2×CLEAR</t> (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .
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    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of <t>2×CLEAR</t> (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .
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    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of <t>2×CLEAR</t> (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .
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    Image Search Results


    APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of 2×CLEAR (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .

    Journal: The Journal of Cell Biology

    Article Title: ATG conjugation–dependent/independent mechanisms underlie lysosomal stress–induced TFEB regulation

    doi: 10.1083/jcb.202307079

    Figure Lengend Snippet: APEX1 is required for TFEB expression and stability. (A) Quantification of the relative expression of TFEB, TFE3, and MITF by qPCR in HeLa cells transfected with siLuc or si APEX1 under nutrient conditions ( n = 3 biologically independent samples). (B) Representative immunoblots of TFEB, p-TFEB (Ser211), APEX1, and GAPDH in WT HeLa cells transfected with EGFP stop (negative control) or EGFP::APEX1, either untreated (control) or treated with 1 mM LLOMe for 1 h. (C) Quantification of 2×CLEAR (TFEB RE)-luciferase reporter assays in HeLa cells stably expressing mNG or APEX1::mNG transfected under 1 mM LLOMe treatment for 3-h conditions ( n = 3 biologically independent samples). (D) Representative immunoblots of TFEB, APEX1, and GAPDH in WT HeLa cells that were either untreated (control) or treated with MG132 for 3 h. Cells were transfected with siLuc or si APEX1 . (E) Quantification of the image data shown in D ( n = 3 biologically independent samples). (F) Representative immunoblots of TFEB and GAPDH in WT HeLa cells treated with chloroquine for 4 h. Cells were transfected with si APEX1 . (G) Quantification of image data shown in F ( n = 3 biologically independent samples). (H) Representative immunofluorescence images of HeLa cells stained using an antibody against endogenous γH2AX (green). Cells were transfected with siLuc or si APEX1 . (I) Representative immunoblots of γH2AX and GAPDH in WT HeLa cells that were transfected with siLuc or si APEX1 . (J) Quantification of image data shown in I ( n = 3 biologically independent samples). (K) Schematic representation of APEX1 mutant constructs. (L) Representative fluorescence images of HeLa cells stably expressing mNG::APEX1 WT or mNG::APEX1 mutants. (M) Results of the PLA using WT HeLa cells stably expressing mNG::stop, mNG::APEX1 WT, or mNG::APEX1 mutants. Cells were transfected with si APEX1 . The indicated plasmids were transiently transfected and were either untreated (control) or treated with 1 mM LLOMe for 1 h followed by the PLA procedure. (H, L, and M) Scale bars, 50 μm. (E) P values were determined by ANOVA with Tukey’s multiple comparison test; *P < 0.05. (A and C) P values were determined by a t test; *P < 0.05, **P < 0.01. Source data are available for this figure: .

    Article Snippet: Briefly, 2×CLEAR (TFEB RE)-luciferase reporters (plasmid #81120; Addgene) and plasmid encoding pMCX-β-galactosidase (gift from Dr. H. Ogawa, The Research foundation for Microbial disease of Osaka University, Suita, Japan) were cotransfected into HeLa cells.

    Techniques: Expressing, Transfection, Western Blot, Negative Control, Control, Luciferase, Stable Transfection, Immunofluorescence, Staining, Mutagenesis, Construct, Fluorescence, Comparison