c ebpβ 3 Search Results


c/ebpβ3′ -luciferase constructs ( c/ebpβ -luc  (Promega)
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Promega c/ebpβ3′ -luciferase constructs ( c/ebpβ -luc
(A) Immunofluorescence analysis of Hoxc8 in the mouse SVF cells derived from inguinal WAT. The cells were untreated (Undifferentiated) or induced to undergo differentiation (Adipogenic induction). The lipid droplets and nuclei were counterstained with Bodipy and DAPI, respectively. Scale bars indicate 30 µm. (B) Immunoblots of Hoxc8 in the mouse SVF cells left untreated or induced to undergo differentiation. β-actin served as a loading control. (C) Upper, the UCP1 expression in the differentiated mouse SVF cells. Scale bars indicate 30 µm. Lower, the fold increase of mRNA expression levels of Ucp1 and Ucp2 in the mouse WAT-SVF cells induced to undergo differentiation. The results were normalized to β-actin . (D) The expression of Pgc-1α and <t>C/EBPβ</t> induced during the differentiation of mouse SVF cells. The results were normalized to β-actin . The data are presented as means ± SEM; * p <0.05. (E) Western blot analysis in different fat depots from mice treated with or without CL-316,243, a β3-adrenergic receptor agonist. ingWAT, epiWAT, and iBAT denote inguinal WAT, epididymal WAT, and interscapular BAT, respectively. (F) Western blot analysis of Hoxc8 in SVF cells and ingWAT of mice treated with CL-316,243 (CL) or saline.
C/Ebpβ3′ Luciferase Constructs ( C/Ebpβ Luc, 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
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(A) Immunofluorescence analysis of Hoxc8 in the mouse SVF cells derived from inguinal WAT. The cells were untreated (Undifferentiated) or induced to undergo differentiation (Adipogenic induction). The lipid droplets and nuclei were counterstained with Bodipy and DAPI, respectively. Scale bars indicate 30 µm. (B) Immunoblots of Hoxc8 in the mouse SVF cells left untreated or induced to undergo differentiation. β-actin served as a loading control. (C) Upper, the UCP1 expression in the differentiated mouse SVF cells. Scale bars indicate 30 µm. Lower, the fold increase of mRNA expression levels of Ucp1 and Ucp2 in the mouse WAT-SVF cells induced to undergo differentiation. The results were normalized to β-actin . (D) The expression of Pgc-1α and C/EBPβ induced during the differentiation of mouse SVF cells. The results were normalized to β-actin . The data are presented as means ± SEM; * p <0.05. (E) Western blot analysis in different fat depots from mice treated with or without CL-316,243, a β3-adrenergic receptor agonist. ingWAT, epiWAT, and iBAT denote inguinal WAT, epididymal WAT, and interscapular BAT, respectively. (F) Western blot analysis of Hoxc8 in SVF cells and ingWAT of mice treated with CL-316,243 (CL) or saline.

Journal: PLoS Biology

Article Title: Essential Role for miR-196a in Brown Adipogenesis of White Fat Progenitor Cells

doi: 10.1371/journal.pbio.1001314

Figure Lengend Snippet: (A) Immunofluorescence analysis of Hoxc8 in the mouse SVF cells derived from inguinal WAT. The cells were untreated (Undifferentiated) or induced to undergo differentiation (Adipogenic induction). The lipid droplets and nuclei were counterstained with Bodipy and DAPI, respectively. Scale bars indicate 30 µm. (B) Immunoblots of Hoxc8 in the mouse SVF cells left untreated or induced to undergo differentiation. β-actin served as a loading control. (C) Upper, the UCP1 expression in the differentiated mouse SVF cells. Scale bars indicate 30 µm. Lower, the fold increase of mRNA expression levels of Ucp1 and Ucp2 in the mouse WAT-SVF cells induced to undergo differentiation. The results were normalized to β-actin . (D) The expression of Pgc-1α and C/EBPβ induced during the differentiation of mouse SVF cells. The results were normalized to β-actin . The data are presented as means ± SEM; * p <0.05. (E) Western blot analysis in different fat depots from mice treated with or without CL-316,243, a β3-adrenergic receptor agonist. ingWAT, epiWAT, and iBAT denote inguinal WAT, epididymal WAT, and interscapular BAT, respectively. (F) Western blot analysis of Hoxc8 in SVF cells and ingWAT of mice treated with CL-316,243 (CL) or saline.

Article Snippet: The C/EBPβ3′ -luciferase constructs ( C/EBPβ -Luc) were generated by cloning the 3′ sequence of the human C/EBPβ gene (+1,021 to +1,837) into the downstream of luciferase gene in pGL3 promoter plasmid (Promega).

Techniques: Immunofluorescence, Derivative Assay, Western Blot, Expressing

(A) The ChIP analysis in 3T3-L1 preadipocytes expressing Flag- Hoxc8 in mouse C/EBPβ locus. H1foo is an oocyte-specific gene and served as a negative control. The numbers 1–4 in C/EBPβ correspond to 1–4 in (B), respectively. (B) The interspecies conservation of the mouse C/EBPβ 3′. The data obtained from the UCSC Genome Browser map. (C) The ChIP analysis in the human WAT-progenitor cells in human C/EBPβ locus. Osteopontin ( OPN ) served as a positive control. (D) The interspecies conservation and location of ChIP primers used in (C) in the human C/EBPβ locus. The data obtained from the UCSC Genome Browser map. (E) Luciferase reporter assay to assess the transcriptional activity of C/EBPβ 3′ sequence inserted into the 3′ end of the luciferase gene. The activity was measured in 3T3-L1 preadipocytes left untreated (Untreated) or induced to undergo adipogenesis (induction). * p <0.05. ♯ p <0.05. (F) Luciferase reporter activity of C/EBPβ 3′ sequence measured in 3T3-L1 preadipocytes transfected with Hoxc8 , homeodomain-mutated Hoxc8 (HDm), or control vector. (G) Luciferase reporter activity in 3T3-L1 preadipocytes transfected with Hoxc8, HDm, or control vector in the presence of trichostatin A, a histone deacetylase (HDAC) inhibitor. (H) Immunoprecipitation in 3T3-L1 preadipocytes stably expressing Flag-Hoxc8. The immunoblot analysis was performed after immunoprecipitation with anti-Flag antibody. The white dot indicates a non-specific band. (I) Immunoprecipitation in 3T3-L1 preadipocytes stably expressing Flag-HDm. The immunoblot analysis was performed after immunoprecipitation with anti-Flag antibody. The white dot indicates a non-specific band. (J) The immunoblot of HDAC3 in the 3T3-L1 preadipocytes transfected with siRNA against HDAC3 . (K) Luciferase reporter activity in 3T3-L1 preadipocytes transfected with Hoxc8 and siRNA against HDAC3 . (L) The mRNA expression levels in the human WAT-progenitor cells stably expressing HOXC8 followed by transfection with C/EBPβ or EGFP and adipogenic induction. All data are presented as means ± SEM. * p <0.05. n.s., not significant.

Journal: PLoS Biology

Article Title: Essential Role for miR-196a in Brown Adipogenesis of White Fat Progenitor Cells

doi: 10.1371/journal.pbio.1001314

Figure Lengend Snippet: (A) The ChIP analysis in 3T3-L1 preadipocytes expressing Flag- Hoxc8 in mouse C/EBPβ locus. H1foo is an oocyte-specific gene and served as a negative control. The numbers 1–4 in C/EBPβ correspond to 1–4 in (B), respectively. (B) The interspecies conservation of the mouse C/EBPβ 3′. The data obtained from the UCSC Genome Browser map. (C) The ChIP analysis in the human WAT-progenitor cells in human C/EBPβ locus. Osteopontin ( OPN ) served as a positive control. (D) The interspecies conservation and location of ChIP primers used in (C) in the human C/EBPβ locus. The data obtained from the UCSC Genome Browser map. (E) Luciferase reporter assay to assess the transcriptional activity of C/EBPβ 3′ sequence inserted into the 3′ end of the luciferase gene. The activity was measured in 3T3-L1 preadipocytes left untreated (Untreated) or induced to undergo adipogenesis (induction). * p <0.05. ♯ p <0.05. (F) Luciferase reporter activity of C/EBPβ 3′ sequence measured in 3T3-L1 preadipocytes transfected with Hoxc8 , homeodomain-mutated Hoxc8 (HDm), or control vector. (G) Luciferase reporter activity in 3T3-L1 preadipocytes transfected with Hoxc8, HDm, or control vector in the presence of trichostatin A, a histone deacetylase (HDAC) inhibitor. (H) Immunoprecipitation in 3T3-L1 preadipocytes stably expressing Flag-Hoxc8. The immunoblot analysis was performed after immunoprecipitation with anti-Flag antibody. The white dot indicates a non-specific band. (I) Immunoprecipitation in 3T3-L1 preadipocytes stably expressing Flag-HDm. The immunoblot analysis was performed after immunoprecipitation with anti-Flag antibody. The white dot indicates a non-specific band. (J) The immunoblot of HDAC3 in the 3T3-L1 preadipocytes transfected with siRNA against HDAC3 . (K) Luciferase reporter activity in 3T3-L1 preadipocytes transfected with Hoxc8 and siRNA against HDAC3 . (L) The mRNA expression levels in the human WAT-progenitor cells stably expressing HOXC8 followed by transfection with C/EBPβ or EGFP and adipogenic induction. All data are presented as means ± SEM. * p <0.05. n.s., not significant.

Article Snippet: The C/EBPβ3′ -luciferase constructs ( C/EBPβ -Luc) were generated by cloning the 3′ sequence of the human C/EBPβ gene (+1,021 to +1,837) into the downstream of luciferase gene in pGL3 promoter plasmid (Promega).

Techniques: Expressing, Negative Control, Positive Control, Luciferase, Reporter Assay, Activity Assay, Sequencing, Transfection, Plasmid Preparation, Histone Deacetylase Assay, Immunoprecipitation, Stable Transfection, Western Blot

Cold temperatures or β3-adrenergic stimulations induce miR-196a in the WAT-resident progenitor cells in mice. miR-196a post-transcriptionally suppress Hoxc8 , which is one of the white-fat genes. The direct target of Hoxc8 is C/EBPβ , a master switch of brown adipogenesis that provokes brown fat gene program in the WAT-progenitor cells.

Journal: PLoS Biology

Article Title: Essential Role for miR-196a in Brown Adipogenesis of White Fat Progenitor Cells

doi: 10.1371/journal.pbio.1001314

Figure Lengend Snippet: Cold temperatures or β3-adrenergic stimulations induce miR-196a in the WAT-resident progenitor cells in mice. miR-196a post-transcriptionally suppress Hoxc8 , which is one of the white-fat genes. The direct target of Hoxc8 is C/EBPβ , a master switch of brown adipogenesis that provokes brown fat gene program in the WAT-progenitor cells.

Article Snippet: The C/EBPβ3′ -luciferase constructs ( C/EBPβ -Luc) were generated by cloning the 3′ sequence of the human C/EBPβ gene (+1,021 to +1,837) into the downstream of luciferase gene in pGL3 promoter plasmid (Promega).

Techniques: