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pcdna3-flag-foxa2  (GenScript corporation)

 
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    GenScript corporation pcdna3-flag-foxa2
    (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) <t>Wild</t> <t>type</t> SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.
    Pcdna3 Flag Foxa2, supplied by GenScript corporation, 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/result/pcdna3-flag-foxa2/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    pcdna3-flag-foxa2 - by Bioz Stars, 2026-03
    90/100 stars

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    1) Product Images from "SIRT1 Mediates FOXA2 Breakdown by Deacetylation in a Nutrient-Dependent Manner"

    Article Title: SIRT1 Mediates FOXA2 Breakdown by Deacetylation in a Nutrient-Dependent Manner

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0098438

    (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) Wild type SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.
    Figure Legend Snippet: (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) Wild type SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.

    Techniques Used: Transfection, Lysis, Western Blot, Control, Mutagenesis, Immunoprecipitation, Activity Assay, Expressing, Cell Culture



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    pcdna3-flag-foxa2  (GenScript corporation)
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    GenScript corporation pcdna3-flag-foxa2
    (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) <t>Wild</t> <t>type</t> SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.
    Pcdna3 Flag Foxa2, supplied by GenScript corporation, 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/result/pcdna3-flag-foxa2/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    pcdna3-flag-foxa2 - by Bioz Stars, 2026-03
    90/100 stars
    		  Buy from Supplier

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    (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) Wild type SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.

    Journal: PLoS ONE

    Article Title: SIRT1 Mediates FOXA2 Breakdown by Deacetylation in a Nutrient-Dependent Manner

    doi: 10.1371/journal.pone.0098438

    Figure Lengend Snippet: (A) SIRT1 affects FOXA2 protein stability. HEK293T cells were transfected with FLAG-FOXA2 in the presence of 20 mM NAM or vehicle for 16 hours prior to cell lysis. Western Blots were probed for FLAG, and actin as a loading control. (B) Wild type SIRT1 and mutant SIRT1 H363Y similarly interact with FOXA2. HEK293T cells were transfected with FLAG-FOXA2 alone or in combination with either wild-type (wt) or the catalytically impaired MYC-SIRT1-H363Y mutant. FOXA2 and SIRT1 were co-immunoprecipitated with anti-FLAG-M2 beads and protein levels were assessed by Western Blot analysis probed with the indicated antibodies. (C) SIRT1 catalytic activity regulates FOXA2 stability. HEK293T cells were transfected and subjected to co-immunoprecipitation as in (B). Protein expression and FOXA2 acetylation level were assessed by Western Blot using the indicated antibodies. (D) Acetylation-impaired FLAG-12K-R-FOXA2 mutant showed reduced acetylation and protein stability. HEK293T cells were transfected with either FLAG-FOXA2 or FLAG-12K-R-FOXA2 and cultured as in (A). After FLAG-immunoprecipitation, protein levels were assessed by Western Blot probed with the indicated antibodies. (E) FOXA2 is subjected to proteasomal degradation in a time-dependent manner. HEK293T cells were transfected with FLAG-FOXA2 and supplemented with proteasome inhibitor MG132 (20 µM) or vehicle-containing medium for 3 or 6 hours. FOXA2 protein levels were assessed by Western Blot, and actin as a loading control. (F) SIRT1 mediates endogenous FOXA2 breakdown. HepG2 cells were transfected with MYC-SIRT1 and supplemented with 20 µM MG132- or vehicle-containing medium for 3 hours. (G) NAM preserves stability of FOXA2 upon abrogation of protein translation. HEK293T cells were transfected with FLAG-FOXA2 and supplemented 20 mM NAM or vehicle for 16 hours, followed by addition of cycloheximide (5 µg/ml) or vehicle-containing medium for 4 or 8 hours. (E–G) show a representative experiment from three independent experiments, and quantification of FOXA2 protein levels was performed by normalizing to actin protein levels. (H) 12K-R FOXA2 mutant has a higher level of poly-ubiquitylation. HEK293T cells were transfected with FLAG-FOXA2 or FLAG-12K-R-FOXA2. After culture of 20 µM MG132 for 3 hours, FOXA2 was immunoprecipitated with FLAG-M2 beads and immunoprecipitated proteins were assessed by Western Blot probed with the indicated antibodies. (I) Nutrient-dependent regulation of Foxa2 acetylation level in murine livers. Murine Foxa2 was immunoprecipitated from whole liver protein lysates from mice fed ad libitum (AL) or starved (ST) to 25% of ad libitum levels. Western Blots were probed with the indicated antibodies. The asterisk (*) marks a co-eluting acetylated protein of unknown identity.

    Article Snippet: Wild type pcDNA3-FLAG-FOXA2 was synthesized by GenScript (Piscataway, NJ, USA). pRSV-MYC-SIRT1 and pRSV-MYC-SIRT1 H363Y were kind gifts from Prof. P. Coffer. pGL3-G6pase-Luc ( G6pase promoter [−1227 to +57]) was a kind gift from Dr. D. Schmoll and pGL3-CPT1a-Luc ( Cpt1a promoter [−2432 to +1]) was a kind gift from Prof. M. Negishi .

    Techniques: Transfection, Lysis, Western Blot, Control, Mutagenesis, Immunoprecipitation, Activity Assay, Expressing, Cell Culture