3t3-l1 mature adipocytes Search Results


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    ATCC mature adipocytes 3t3 l1 preadipocytes
    The components IPTP and TPP induce endogenous expression and recruitment of PPARγ to the aP2 enhancer region. <t>3T3-L1</t> preadipocytes cells exposed to the differentiation cocktail for 6 days were used for the ChIP assay to measure the endogenous expression and recruitment of PPARγ to the aP2 enhancer region by RT-PCR. Positive controls dexamethasone (DEX) and troglitazone (TROG) were used and depicted in ( A ). ( B ) PPARγ recruitment to the aP2 enhancer region after exposure to 20 μM TPP and 10 μM IPTP was determined. Data represents the mean of n = 6 independent replicates ± SEM. Data were relative to 100% input DNA and significance was determined relative to control PPARγ recruitment using a one-way ANOVA followed by Tukey’s post-hoc analysis (* P
    Mature Adipocytes 3t3 L1 Preadipocytes, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    The components IPTP and TPP induce endogenous expression and recruitment of PPARγ to the aP2 enhancer region. 3T3-L1 preadipocytes cells exposed to the differentiation cocktail for 6 days were used for the ChIP assay to measure the endogenous expression and recruitment of PPARγ to the aP2 enhancer region by RT-PCR. Positive controls dexamethasone (DEX) and troglitazone (TROG) were used and depicted in ( A ). ( B ) PPARγ recruitment to the aP2 enhancer region after exposure to 20 μM TPP and 10 μM IPTP was determined. Data represents the mean of n = 6 independent replicates ± SEM. Data were relative to 100% input DNA and significance was determined relative to control PPARγ recruitment using a one-way ANOVA followed by Tukey’s post-hoc analysis (* P

    Journal: PLoS ONE

    Article Title: Firemaster® 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor (Pparγ) on the adipocyte protein 2 (aP2) promoter

    doi: 10.1371/journal.pone.0175855

    Figure Lengend Snippet: The components IPTP and TPP induce endogenous expression and recruitment of PPARγ to the aP2 enhancer region. 3T3-L1 preadipocytes cells exposed to the differentiation cocktail for 6 days were used for the ChIP assay to measure the endogenous expression and recruitment of PPARγ to the aP2 enhancer region by RT-PCR. Positive controls dexamethasone (DEX) and troglitazone (TROG) were used and depicted in ( A ). ( B ) PPARγ recruitment to the aP2 enhancer region after exposure to 20 μM TPP and 10 μM IPTP was determined. Data represents the mean of n = 6 independent replicates ± SEM. Data were relative to 100% input DNA and significance was determined relative to control PPARγ recruitment using a one-way ANOVA followed by Tukey’s post-hoc analysis (* P

    Article Snippet: Differentiation of 3T3-L1 preadipocytes into mature adipocytes 3T3-L1 preadipocytes (ATCC® CL-173™ ) purchased from the American Type Culture Collection were differentiated as previously described [ ].

    Techniques: Expressing, Chromatin Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction

    Temporal expression of transcription factors and terminal differentiation markers after exposure to Firemaster ® 550 and its components. mRNA expression levels were determined in 3T3-L1 preadipocytes 2, 4, 6, and 9 days post-treatment with the differentiation cocktail consisting of IBMX, insulin, and either 100 μM FM550, 10 μM IPTP, 20 μM TPP, or 20 μM TBPH. After the indicated time-points, RNA was extracted and reverse transcribed, and we measured the levels of ( A ) Pparγ , ( B ) Cebpα , ( C ) aP2 , ( D ) Lpl , and ( E ) Plin , which were normalized to β-actin levels and relative to time-matched vehicle control (MI). Data represent the mean ± S.E.M ( n = 3–5). * P

    Journal: PLoS ONE

    Article Title: Firemaster® 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor (Pparγ) on the adipocyte protein 2 (aP2) promoter

    doi: 10.1371/journal.pone.0175855

    Figure Lengend Snippet: Temporal expression of transcription factors and terminal differentiation markers after exposure to Firemaster ® 550 and its components. mRNA expression levels were determined in 3T3-L1 preadipocytes 2, 4, 6, and 9 days post-treatment with the differentiation cocktail consisting of IBMX, insulin, and either 100 μM FM550, 10 μM IPTP, 20 μM TPP, or 20 μM TBPH. After the indicated time-points, RNA was extracted and reverse transcribed, and we measured the levels of ( A ) Pparγ , ( B ) Cebpα , ( C ) aP2 , ( D ) Lpl , and ( E ) Plin , which were normalized to β-actin levels and relative to time-matched vehicle control (MI). Data represent the mean ± S.E.M ( n = 3–5). * P

    Article Snippet: Differentiation of 3T3-L1 preadipocytes into mature adipocytes 3T3-L1 preadipocytes (ATCC® CL-173™ ) purchased from the American Type Culture Collection were differentiated as previously described [ ].

    Techniques: Expressing

    Lipid accumulation after exposure to Firemaster ® 550 and its components using 3T3-L1 preadipocytes. ( A ) Preadipocytes were differentiated as described in the Materials and Methods with vehicle with IBMX and insulin (MI) or increasing amounts of Firemaster ® 550 (0.1–200 μM) or with the FM550 components ( B ) IPTP, TPP, TBPH, TBB, and DBB (0.1–20 μM) and ( C ) 250 nM dexamethasone (MID), or 5 μM troglitazone (MIT). After 9 days lipid accumulation was visualized using Nile Red staining and then quantified. Lipid accumulation was normalized to DAPI staining and relative to vehicle-treated cells. The photographs are representative data obtained from experiments run in parallel with all the chemicals. Data represent mean ± SEM for n = 3–5 independent experiments performed in triplicate. Statistical significance * P

    Journal: PLoS ONE

    Article Title: Firemaster® 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor (Pparγ) on the adipocyte protein 2 (aP2) promoter

    doi: 10.1371/journal.pone.0175855

    Figure Lengend Snippet: Lipid accumulation after exposure to Firemaster ® 550 and its components using 3T3-L1 preadipocytes. ( A ) Preadipocytes were differentiated as described in the Materials and Methods with vehicle with IBMX and insulin (MI) or increasing amounts of Firemaster ® 550 (0.1–200 μM) or with the FM550 components ( B ) IPTP, TPP, TBPH, TBB, and DBB (0.1–20 μM) and ( C ) 250 nM dexamethasone (MID), or 5 μM troglitazone (MIT). After 9 days lipid accumulation was visualized using Nile Red staining and then quantified. Lipid accumulation was normalized to DAPI staining and relative to vehicle-treated cells. The photographs are representative data obtained from experiments run in parallel with all the chemicals. Data represent mean ± SEM for n = 3–5 independent experiments performed in triplicate. Statistical significance * P

    Article Snippet: Differentiation of 3T3-L1 preadipocytes into mature adipocytes 3T3-L1 preadipocytes (ATCC® CL-173™ ) purchased from the American Type Culture Collection were differentiated as previously described [ ].

    Techniques: Staining

    The effects of selective PPARγ antagonist GW9662 on aP2 mRNA expression and lipid accumulation in the presence of troglitazone, TPP, IPTP, and TBPH. Murine 3T3-L1 preadipocytes were induced to differentiate in the presence of 500 μM IBMX (M), 100 nM insulin (I), supplemented with indicated treatments: 5 μM troglitazone (TROG), 0.25 μM dexamethasone (DEX), 20 μM TPP, 10 μM IPTP, or 20 μM TBPH with either solvent control (Veh) or PPARγ inhibitor GW9662 (GW) twice daily. ( A ) At day 6 of differentiation mRNA expression levels of aP2 were quantified by RT-qPCR normalized to MI control conditions. ( B-E ) At days 2, 4, 6, and 9 lipid accumulation was visualized using Nile Red staining and then quantified. Lipid accumulation was normalized to DAPI staining. Data represent mean ± SEM for n = 3 independent experiments. * denotes p

    Journal: PLoS ONE

    Article Title: Firemaster® 550 and its components isopropylated triphenyl phosphate and triphenyl phosphate enhance adipogenesis and transcriptional activity of peroxisome proliferator activated receptor (Pparγ) on the adipocyte protein 2 (aP2) promoter

    doi: 10.1371/journal.pone.0175855

    Figure Lengend Snippet: The effects of selective PPARγ antagonist GW9662 on aP2 mRNA expression and lipid accumulation in the presence of troglitazone, TPP, IPTP, and TBPH. Murine 3T3-L1 preadipocytes were induced to differentiate in the presence of 500 μM IBMX (M), 100 nM insulin (I), supplemented with indicated treatments: 5 μM troglitazone (TROG), 0.25 μM dexamethasone (DEX), 20 μM TPP, 10 μM IPTP, or 20 μM TBPH with either solvent control (Veh) or PPARγ inhibitor GW9662 (GW) twice daily. ( A ) At day 6 of differentiation mRNA expression levels of aP2 were quantified by RT-qPCR normalized to MI control conditions. ( B-E ) At days 2, 4, 6, and 9 lipid accumulation was visualized using Nile Red staining and then quantified. Lipid accumulation was normalized to DAPI staining. Data represent mean ± SEM for n = 3 independent experiments. * denotes p

    Article Snippet: Differentiation of 3T3-L1 preadipocytes into mature adipocytes 3T3-L1 preadipocytes (ATCC® CL-173™ ) purchased from the American Type Culture Collection were differentiated as previously described [ ].

    Techniques: Expressing, Quantitative RT-PCR, Staining

    SENP1 deletion alters pancreatic adipocyte phenotype and augments NF-κB-dependent inflammation. ( a , b ) Pancreatic adipose was collected from 7 and 14 weeks old Ctrl and SENP1-aP2KO male (male, n =6). Morphology was visualized by haematoxylin and eosin stain staining. Scale bar, 20 μm ( a ). Cell sizes were quantified in ( b ). Three sections from each adipose tissue. Data are representative for three independent experiments. ( c ) Transcript levels of adipocyte differentiation markers (fatty acid synthase, adipose triglyceride lipase and lipoprotein lipase) in PATs were quantified by quantitative PCR with reverse transcription with GAPDH for normalization. n =6, male for each group. ( d ) Increased IKK-NF-κB activities in SENP1-aP2KO PATs. A representative western blot was from Ctrl and SENP1-aP2KO mice at the age of 7 weeks. Data are representative from three independent experiments is shown ( n =6, male). Relative protein levels were quantified from three blots by taking Ctrl as 1.0 ( n =6, male). ( e – h ) NF-κB activation is specifically detected in PATs of SENP1-aP2KO mice. ( e ) Phosphor-p65 staining (red) in PATs but not in pancreas. ( f ) Co-immunostaining of phosphor-p65 (green) and adipocyte marker FABP4 (red). ( g ) High-power images show co-staining of phosphor-p65 (red) in the nucleus of FABP4 + adipocytes (green). ( h ) Co-staining of phosphor-p65 (green) with APC-conjugated adipocyte marker FABP4 (green; yellow arrowheads), but not with macrophage marker F4/80 (red; white arrowheads). Representative images are from one of three sections from SENP1-aP2KO PATs and n =3 male mice. Scale bar, 20 μm. ( i , j ) ChIP assay. ChIP assays with p65/RelA antibody were performed in the adipocyte isolated from Ctrl ( n =6, male) and SENP1-aP2KO mice ( n =6, male) with an IgG isotype as a control. Bindings of p65/RelA to the IL-6, IL-1β and TNF-α gene promoters were quantified with the ratio of IP/input for each promoter ( j ). ( k , l ) Effects of SENP1 knockdown on NF-κB and cytokine expression in adipocytes. 3T3-L1 adipocytes were transfected with control or SENP1-specific siRNA for 24 h. IKK-NF-κB p65/RelA signalling molecules were detected by western blotting. A representative blot from three experiments is shown and protein levels are quantified by taking Ctrl as 1.0 ( k ). Protein levels of IL-6, TNF-α, IFN-γ and IL-1β levels present in adipocyte cultures were detected with ELISA after 24-h culture ( l ). All data are means±s.e.m. of three independent experiments. The two-tailed Student's paired t -test was used for the statistical analysis. * P

    Journal: Nature Communications

    Article Title: SENP1-mediated NEMO deSUMOylation in adipocytes limits inflammatory responses and type-1 diabetes progression

    doi: 10.1038/ncomms9917

    Figure Lengend Snippet: SENP1 deletion alters pancreatic adipocyte phenotype and augments NF-κB-dependent inflammation. ( a , b ) Pancreatic adipose was collected from 7 and 14 weeks old Ctrl and SENP1-aP2KO male (male, n =6). Morphology was visualized by haematoxylin and eosin stain staining. Scale bar, 20 μm ( a ). Cell sizes were quantified in ( b ). Three sections from each adipose tissue. Data are representative for three independent experiments. ( c ) Transcript levels of adipocyte differentiation markers (fatty acid synthase, adipose triglyceride lipase and lipoprotein lipase) in PATs were quantified by quantitative PCR with reverse transcription with GAPDH for normalization. n =6, male for each group. ( d ) Increased IKK-NF-κB activities in SENP1-aP2KO PATs. A representative western blot was from Ctrl and SENP1-aP2KO mice at the age of 7 weeks. Data are representative from three independent experiments is shown ( n =6, male). Relative protein levels were quantified from three blots by taking Ctrl as 1.0 ( n =6, male). ( e – h ) NF-κB activation is specifically detected in PATs of SENP1-aP2KO mice. ( e ) Phosphor-p65 staining (red) in PATs but not in pancreas. ( f ) Co-immunostaining of phosphor-p65 (green) and adipocyte marker FABP4 (red). ( g ) High-power images show co-staining of phosphor-p65 (red) in the nucleus of FABP4 + adipocytes (green). ( h ) Co-staining of phosphor-p65 (green) with APC-conjugated adipocyte marker FABP4 (green; yellow arrowheads), but not with macrophage marker F4/80 (red; white arrowheads). Representative images are from one of three sections from SENP1-aP2KO PATs and n =3 male mice. Scale bar, 20 μm. ( i , j ) ChIP assay. ChIP assays with p65/RelA antibody were performed in the adipocyte isolated from Ctrl ( n =6, male) and SENP1-aP2KO mice ( n =6, male) with an IgG isotype as a control. Bindings of p65/RelA to the IL-6, IL-1β and TNF-α gene promoters were quantified with the ratio of IP/input for each promoter ( j ). ( k , l ) Effects of SENP1 knockdown on NF-κB and cytokine expression in adipocytes. 3T3-L1 adipocytes were transfected with control or SENP1-specific siRNA for 24 h. IKK-NF-κB p65/RelA signalling molecules were detected by western blotting. A representative blot from three experiments is shown and protein levels are quantified by taking Ctrl as 1.0 ( k ). Protein levels of IL-6, TNF-α, IFN-γ and IL-1β levels present in adipocyte cultures were detected with ELISA after 24-h culture ( l ). All data are means±s.e.m. of three independent experiments. The two-tailed Student's paired t -test was used for the statistical analysis. * P

    Article Snippet: Cell lines and isolation of mature adipocytes 3T3-L1, a commonly used preadipocyte mouse cell line, was obtained from ATCC (CL-173) and cultured under DMEM media supplemented with 2 mM glutamine and 10% calf serum.

    Techniques: H&E Stain, Staining, Real-time Polymerase Chain Reaction, Western Blot, Mouse Assay, Activation Assay, Immunostaining, Marker, Chromatin Immunoprecipitation, Isolation, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Two Tailed Test