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fgf21 promoter driven luciferase reporter construct (Addgene inc)

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Addgene inc fgf21 promoter driven luciferase reporter construct

Fig. 1. Effects of 3-O-acetyloleanolic acid isolated from Forsythiae Fructus on the secretion of <t>FGF21</t> in C2C12 myotubes. (A) Outline of the extraction and isolation of 3-O-acetyloleanolic acid from Forsythiae Fructus. (B) Structure of 3-O-acetyloleanolic acid. (C) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid isolated from Forsythiae Fructus in C2C12 myotubes. Each value is the mean ± SEM of four separate experiments. The double asterisk indicates P < 0.01.

Fgf21 Promoter Driven Luciferase Reporter Construct, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/fgf21+promoter+driven+luciferase+reporter+construct/pGL3-mFgf21pro1-luc+(Plasmid+%23101797)/pm34449315-75-1-40
Average 90 stars, based on 3 article reviews

fgf21 promoter driven luciferase reporter construct - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "Transmembrane G protein-coupled receptor 5 signaling stimulates fibroblast growth factor 21 expression concomitant with up-regulation of the transcription factor nuclear receptor Nr4a1."

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Fig. 1. Effects of 3-O-acetyloleanolic acid isolated from Forsythiae Fructus on the secretion of FGF21 in C2C12 myotubes. (A) Outline of the extraction and isolation of 3-O-acetyloleanolic acid from Forsythiae Fructus. (B) Structure of 3-O-acetyloleanolic acid. (C) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid isolated from Forsythiae Fructus in C2C12 myotubes. Each value is the mean ± SEM of four separate experiments. The double asterisk indicates P < 0.01.

Figure Legend Snippet: Fig. 1. Effects of 3-O-acetyloleanolic acid isolated from Forsythiae Fructus on the secretion of FGF21 in C2C12 myotubes. (A) Outline of the extraction and isolation of 3-O-acetyloleanolic acid from Forsythiae Fructus. (B) Structure of 3-O-acetyloleanolic acid. (C) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid isolated from Forsythiae Fructus in C2C12 myotubes. Each value is the mean ± SEM of four separate experiments. The double asterisk indicates P < 0.01.

Techniques Used: Isolation, Extraction

Fig. 2. 3-O-Acetyloleanolic acid up- regulates FGF21 expression in C2C12 myotubes. (A) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid in a concentration-dependent manner in C2C12 myotubes. Cells were treated with the indicated concentration of 3-O-acetyloleanolic acid for 20 h, then the media were collected and pro cessed for ELISA. Each value represents the mean ± SEM of five separate ex periments. **P < 0.01, comparison with vehicle control. (B) FGF21 mRNA expression was up-regulated by 3-O- acetyloleanolic acid in C2C12 myo tubes. Cells were treated with 30 µM 3- O-acetyloleanolic acid for 2 and 5 h, harvested, and then processed for real- time qRT-PCR. Each value represents the mean ± SEM of five separate ex periments. *P < 0.01; **P < 0.01, com parison with vehicle control.

Figure Legend Snippet: Fig. 2. 3-O-Acetyloleanolic acid up- regulates FGF21 expression in C2C12 myotubes. (A) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid in a concentration-dependent manner in C2C12 myotubes. Cells were treated with the indicated concentration of 3-O-acetyloleanolic acid for 20 h, then the media were collected and pro cessed for ELISA. Each value represents the mean ± SEM of five separate ex periments. **P < 0.01, comparison with vehicle control. (B) FGF21 mRNA expression was up-regulated by 3-O- acetyloleanolic acid in C2C12 myo tubes. Cells were treated with 30 µM 3- O-acetyloleanolic acid for 2 and 5 h, harvested, and then processed for real- time qRT-PCR. Each value represents the mean ± SEM of five separate ex periments. *P < 0.01; **P < 0.01, com parison with vehicle control.

Techniques Used: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison, Control, Quantitative RT-PCR

Fig. 3. 3-O-Acetyloleanolic acid stimulates FGF21 expression in mice. (A) Effect of the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg on FGF21 mRNA levels in soleus muscle. Total RNA was isolated from the soleus muscle of saline-treated and 3-O-acetyloleanolic acid-treated mice. FGF21 mRNA was determined by qRT-PCR using specific primers. Relative mRNA levels were normalized to 18S. Each value is the mean ± SEM of five mice. The single asterisk indicates P < 0.05. (B) 3-O-acetyloleanolic acid at 30 mg/kg increased circulating FGF21. The plasma FGF21 concentration was determined by FGF21-specific ELISA after the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg. Each value represents the mean ± SEM of five mice. The double asterisk in dicates P < 0.01.

Figure Legend Snippet: Fig. 3. 3-O-Acetyloleanolic acid stimulates FGF21 expression in mice. (A) Effect of the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg on FGF21 mRNA levels in soleus muscle. Total RNA was isolated from the soleus muscle of saline-treated and 3-O-acetyloleanolic acid-treated mice. FGF21 mRNA was determined by qRT-PCR using specific primers. Relative mRNA levels were normalized to 18S. Each value is the mean ± SEM of five mice. The single asterisk indicates P < 0.05. (B) 3-O-acetyloleanolic acid at 30 mg/kg increased circulating FGF21. The plasma FGF21 concentration was determined by FGF21-specific ELISA after the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg. Each value represents the mean ± SEM of five mice. The double asterisk in dicates P < 0.01.

Techniques Used: Expressing, Isolation, Saline, Quantitative RT-PCR, Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay

Fig. 5. Role of p38 MAPK in the up-regulation of FGF21 secretion caused by 3-O-acetyloleanolic acid in C2C12 cells. (A) Cells were treated with 30 µM 3-O-ace tyloleanolic acid for 10 min, harvested, and then processed for western blotting analysis of the phosphorylated and total p38 levels in whole cell lysate. Western blots were quantified using ImageJ software (National Institutes of Health). Each value represents the mean ± SEM of four to five separate experiments. **P < 0.01, comparison with vehicle control. (B) Relative luciferase activity of the FGF21 promoter construct (mFgf21pro1-luc) in C2C12 cells following treatment with 3-O- acetyloleanolic acid and co-transfection of the TGR5-expression plasmid. The total amount of DNA transfected was standardized with an empty vector. After transfection, the cells were preincubated with the indicated concentrations of SB203580 for 20 min and then stimulated with 30 μM 3-O-acetyloleanolic acid for 8 h. Luciferase activity was normalized by β-gal and expressed relative to that in vehicle-treated cells. Each value is the mean ± SEM of five independent experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (C) Cells were incubated for 20 h with 3-O- acetyloleanolic acid and SB203580, and FGF21 production was determined by ELISA. Each value represents the mean ± SEM of five separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (D) Cells were incubated for 5 h in 3-O- acetyloleanolic acid and SB203580, and Fgf21 mRNA levels were determined by qRT-PCR. Fgf21 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of five experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations.

Figure Legend Snippet: Fig. 5. Role of p38 MAPK in the up-regulation of FGF21 secretion caused by 3-O-acetyloleanolic acid in C2C12 cells. (A) Cells were treated with 30 µM 3-O-ace tyloleanolic acid for 10 min, harvested, and then processed for western blotting analysis of the phosphorylated and total p38 levels in whole cell lysate. Western blots were quantified using ImageJ software (National Institutes of Health). Each value represents the mean ± SEM of four to five separate experiments. **P < 0.01, comparison with vehicle control. (B) Relative luciferase activity of the FGF21 promoter construct (mFgf21pro1-luc) in C2C12 cells following treatment with 3-O- acetyloleanolic acid and co-transfection of the TGR5-expression plasmid. The total amount of DNA transfected was standardized with an empty vector. After transfection, the cells were preincubated with the indicated concentrations of SB203580 for 20 min and then stimulated with 30 μM 3-O-acetyloleanolic acid for 8 h. Luciferase activity was normalized by β-gal and expressed relative to that in vehicle-treated cells. Each value is the mean ± SEM of five independent experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (C) Cells were incubated for 20 h with 3-O- acetyloleanolic acid and SB203580, and FGF21 production was determined by ELISA. Each value represents the mean ± SEM of five separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (D) Cells were incubated for 5 h in 3-O- acetyloleanolic acid and SB203580, and Fgf21 mRNA levels were determined by qRT-PCR. Fgf21 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of five experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations.

Techniques Used: Western Blot, Software, Comparison, Control, Luciferase, Activity Assay, Construct, Cotransfection, Expressing, Plasmid Preparation, Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Fig. 6. 3-O-acetyloleanolic acid up-regulates FGF21 expression concomitant with the up-regulation of Nr4a1 in C2C12 myotubes. (A) Nr4a1 mRNA expression was up-regulated by 3-O-acetyloleanolic acid in C2C12 myotubes. Cells were treated with 30 µM 3-O-acetyloleanolic acid for 1, 2 and 5 h, harvested, and then processed for real-time qRT-PCR. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison with vehicle control. (B) Cells were incubated for 5 h with 3-O-acetyloleanolic acid and SB203580, and Nr4a1 mRNA levels were determined by qRT-PCR. Nr4a1 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, com parison of SB203580 concentrations. (C) Cells were transfected with a wild-type FGF21 promoter construct (mFgf21pro1-luc) or NBRE-mutated version (NBRE-mut- mFgf21pro1-luc), and relative luciferase activity was measured. Data are means ± SEM of four separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of constructs.

Figure Legend Snippet: Fig. 6. 3-O-acetyloleanolic acid up-regulates FGF21 expression concomitant with the up-regulation of Nr4a1 in C2C12 myotubes. (A) Nr4a1 mRNA expression was up-regulated by 3-O-acetyloleanolic acid in C2C12 myotubes. Cells were treated with 30 µM 3-O-acetyloleanolic acid for 1, 2 and 5 h, harvested, and then processed for real-time qRT-PCR. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison with vehicle control. (B) Cells were incubated for 5 h with 3-O-acetyloleanolic acid and SB203580, and Nr4a1 mRNA levels were determined by qRT-PCR. Nr4a1 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, com parison of SB203580 concentrations. (C) Cells were transfected with a wild-type FGF21 promoter construct (mFgf21pro1-luc) or NBRE-mutated version (NBRE-mut- mFgf21pro1-luc), and relative luciferase activity was measured. Data are means ± SEM of four separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of constructs.

Techniques Used: Expressing, Quantitative RT-PCR, Comparison, Control, Incubation, Transfection, Construct, Luciferase, Activity Assay

Luciferase:

Article Title: Transmembrane G protein-coupled receptor 5 signaling stimulates fibroblast growth factor 21 expression concomitant with up-regulation of the transcription factor nuclear receptor Nr4a1.
Article Snippet: .. The Fgf21 promoter-driven luciferase reporter construct, mFgf21pro1-luc, containing a DNA fragment corresponding to − 1326 to +100 in the region upstream of the murine Fgf21 transcription initiation site linked to the luciferase reporter gene, was a gift from Seiichi Oyadomari (Addgene plasmid #101797; http://n2t.net/addgene:101797; RRID: Addgene_101797) [40]. .. The mutant construct (NBRE-mut-mFgf21pro1-luc) was generated from the mFgf21pro1-luc plasmid by PCR amplification using Prime STAR mutagenesis (Takara) and the following primers: 5′- TTCTAAGAATCCACTCCCCAGCCTCCT-3′ and 5′- AGTGGATTCTTAGAACTGATGTGGCCC-3′.

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Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Figure Lengend Snippet: Fig. 1. Effects of 3-O-acetyloleanolic acid isolated from Forsythiae Fructus on the secretion of FGF21 in C2C12 myotubes. (A) Outline of the extraction and isolation of 3-O-acetyloleanolic acid from Forsythiae Fructus. (B) Structure of 3-O-acetyloleanolic acid. (C) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid isolated from Forsythiae Fructus in C2C12 myotubes. Each value is the mean ± SEM of four separate experiments. The double asterisk indicates P < 0.01.

Article Snippet: The Fgf21 promoter-driven luciferase reporter construct, mFgf21pro1-luc, containing a DNA fragment corresponding to − 1326 to +100 in the region upstream of the murine Fgf21 transcription initiation site linked to the luciferase reporter gene, was a gift from Seiichi Oyadomari (Addgene plasmid #101797; http://n2t.net/addgene:101797; RRID: Addgene_101797) [40].

Techniques: Isolation, Extraction

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Figure Lengend Snippet: Fig. 2. 3-O-Acetyloleanolic acid up- regulates FGF21 expression in C2C12 myotubes. (A) The secretion of FGF21 was increased by 3-O-acetyloleanolic acid in a concentration-dependent manner in C2C12 myotubes. Cells were treated with the indicated concentration of 3-O-acetyloleanolic acid for 20 h, then the media were collected and pro cessed for ELISA. Each value represents the mean ± SEM of five separate ex periments. **P < 0.01, comparison with vehicle control. (B) FGF21 mRNA expression was up-regulated by 3-O- acetyloleanolic acid in C2C12 myo tubes. Cells were treated with 30 µM 3- O-acetyloleanolic acid for 2 and 5 h, harvested, and then processed for real- time qRT-PCR. Each value represents the mean ± SEM of five separate ex periments. *P < 0.01; **P < 0.01, com parison with vehicle control.

Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison, Control, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Figure Lengend Snippet: Fig. 3. 3-O-Acetyloleanolic acid stimulates FGF21 expression in mice. (A) Effect of the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg on FGF21 mRNA levels in soleus muscle. Total RNA was isolated from the soleus muscle of saline-treated and 3-O-acetyloleanolic acid-treated mice. FGF21 mRNA was determined by qRT-PCR using specific primers. Relative mRNA levels were normalized to 18S. Each value is the mean ± SEM of five mice. The single asterisk indicates P < 0.05. (B) 3-O-acetyloleanolic acid at 30 mg/kg increased circulating FGF21. The plasma FGF21 concentration was determined by FGF21-specific ELISA after the intraperitoneal administration of 3-O-acetyloleanolic acid at 30 mg/kg. Each value represents the mean ± SEM of five mice. The double asterisk in dicates P < 0.01.

Techniques: Expressing, Isolation, Saline, Quantitative RT-PCR, Clinical Proteomics, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Figure Lengend Snippet: Fig. 5. Role of p38 MAPK in the up-regulation of FGF21 secretion caused by 3-O-acetyloleanolic acid in C2C12 cells. (A) Cells were treated with 30 µM 3-O-ace tyloleanolic acid for 10 min, harvested, and then processed for western blotting analysis of the phosphorylated and total p38 levels in whole cell lysate. Western blots were quantified using ImageJ software (National Institutes of Health). Each value represents the mean ± SEM of four to five separate experiments. **P < 0.01, comparison with vehicle control. (B) Relative luciferase activity of the FGF21 promoter construct (mFgf21pro1-luc) in C2C12 cells following treatment with 3-O- acetyloleanolic acid and co-transfection of the TGR5-expression plasmid. The total amount of DNA transfected was standardized with an empty vector. After transfection, the cells were preincubated with the indicated concentrations of SB203580 for 20 min and then stimulated with 30 μM 3-O-acetyloleanolic acid for 8 h. Luciferase activity was normalized by β-gal and expressed relative to that in vehicle-treated cells. Each value is the mean ± SEM of five independent experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (C) Cells were incubated for 20 h with 3-O- acetyloleanolic acid and SB203580, and FGF21 production was determined by ELISA. Each value represents the mean ± SEM of five separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations. (D) Cells were incubated for 5 h in 3-O- acetyloleanolic acid and SB203580, and Fgf21 mRNA levels were determined by qRT-PCR. Fgf21 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of five experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of SB203580 concentrations.

Techniques: Western Blot, Software, Comparison, Control, Luciferase, Activity Assay, Construct, Cotransfection, Expressing, Plasmid Preparation, Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

doi: 10.1016/j.biopha.2021.112078

Figure Lengend Snippet: Fig. 6. 3-O-acetyloleanolic acid up-regulates FGF21 expression concomitant with the up-regulation of Nr4a1 in C2C12 myotubes. (A) Nr4a1 mRNA expression was up-regulated by 3-O-acetyloleanolic acid in C2C12 myotubes. Cells were treated with 30 µM 3-O-acetyloleanolic acid for 1, 2 and 5 h, harvested, and then processed for real-time qRT-PCR. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison with vehicle control. (B) Cells were incubated for 5 h with 3-O-acetyloleanolic acid and SB203580, and Nr4a1 mRNA levels were determined by qRT-PCR. Nr4a1 mRNA levels were normalized relative to 18S mRNA levels. Each value represents the mean ± SEM of four separate experiments. *P < 0.05, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, com parison of SB203580 concentrations. (C) Cells were transfected with a wild-type FGF21 promoter construct (mFgf21pro1-luc) or NBRE-mutated version (NBRE-mut- mFgf21pro1-luc), and relative luciferase activity was measured. Data are means ± SEM of four separate experiments. **P < 0.01, comparison of 3-O-acetyloleanolic acid concentrations. ##P < 0.01, comparison of constructs.

Techniques: Expressing, Quantitative RT-PCR, Comparison, Control, Incubation, Transfection, Construct, Luciferase, Activity Assay

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fgf21 promoter driven luciferase reporter construct (Addgene inc)

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1) Product Images from "Transmembrane G protein-coupled receptor 5 signaling stimulates fibroblast growth factor 21 expression concomitant with up-regulation of the transcription factor nuclear receptor Nr4a1."

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