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2×clear tfeb re luciferase reporters  (Addgene inc)


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    Addgene inc 2×clear tfeb re luciferase reporters
    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: .
    2×Clear Tfeb Re Luciferase Reporters, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/2%C3%97clear+tfeb+re+luciferase+reporters/pGL2-2xCLEAR-luciferase+(Plasmid+%2381120)/pmc12396377-219-1-7
    Average 93 stars, based on 4 article reviews
    2×clear tfeb re luciferase reporters - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "ATG conjugation–dependent/independent mechanisms underlie lysosomal stress–induced TFEB regulation"

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

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.202307079

    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: .
    Figure Legend 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: .

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

    Related Articles

    Plasmid Preparation:

    Article Title: ATG conjugation–dependent/independent mechanisms underlie lysosomal stress–induced TFEB regulation
    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. .. After being treated with 1 mM LLOMe for 3 h, cells were lysed at room temperature, and luciferase activity and β-galactosidase activity were measured using a commercially available luciferase assay system (E4030; Promega) and β-galactosidase enzyme assay system (E2000; Promega).



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    2×clear tfeb re luciferase reporters  (Addgene inc)
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    Addgene inc 2×clear tfeb re luciferase reporters
    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: .
    2×Clear Tfeb Re Luciferase Reporters, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/2%C3%97clear+tfeb+re+luciferase+reporters/pGL2-2xCLEAR-luciferase+(Plasmid+%2381120)/pmc12396377-219-1-7
    Average 93 stars, based on 1 article reviews
    2×clear tfeb re luciferase reporters - by Bioz Stars, 2026-09
    93/100 stars
    		  Buy from Supplier

    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