sw 872 Search Results


listed liposarcoma cell line sw872  (ATCC)
95
ATCC listed liposarcoma cell line sw872
Listed Liposarcoma Cell Line Sw872, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sw 872 cells  (CLS Cell Lines Service GmbH)
90
CLS Cell Lines Service GmbH sw 872 cells
Figure 2. In vivo application of [D]-K3H3L9. (a) Eighty-six percent remission in s.c. <t>implanted</t> <t>SW-872</t> liposarcoma xenografts after
Sw 872 Cells, supplied by CLS Cell Lines Service GmbH, 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/product/sw+872/SW-872+Cells/pm20734391-33-0-5
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sw 872 cells - by Bioz Stars, 2026-09
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human liposarcoma cell line sw872  (ATCC)
94
ATCC human liposarcoma cell line sw872
FIG. 2. Increased cellular degradation of 125I-2M* by cells treated with PPAR ligands. A, differentiated primary human adi- pocytes were treated with various concentrations of rosiglitazone in the absence of insulin for 24 h prior to the experiment. The cells were then incubated at 37 °C with HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M* with or without RAP (30 g/ml). At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble material was used as a measure of the degraded protein. The data points represent the means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.009; **, p 0.007; ***, p 0.002. B and C, <t>SW872</t> cells (treated with vehicle, 500 nM rosiglitazone, and 160 M arachidonic acid) were washed at 37 °C to remove any residual fetal calf serum. The cells were incubated at 37 °C for the indicated times in HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M*. At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble ma- terial was used as a measure of the degraded protein. Trichloroacetic acid-soluble material from untreated cells (vehicle) was compared with the trichloroacetic acid-soluble material from cells treated with 500 nM rosiglitazone (B) or 160 M arachidonic acid (C). The data points rep- resent means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.006; **, p 0.03. DMSO, dimethyl sulfoxide.
Human Liposarcoma Cell Line Sw872, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sw+872/SW+872%3B+Liposarcoma%3B+Human/10__1074_slash_jbc__m212989200-73-2-7
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human liposarcoma cell line sw872 - by Bioz Stars, 2026-09
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sw 872 bcrc 60432  (BioResource International Inc)
90
BioResource International Inc sw 872 bcrc 60432
FIG. 2. Increased cellular degradation of 125I-2M* by cells treated with PPAR ligands. A, differentiated primary human adi- pocytes were treated with various concentrations of rosiglitazone in the absence of insulin for 24 h prior to the experiment. The cells were then incubated at 37 °C with HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M* with or without RAP (30 g/ml). At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble material was used as a measure of the degraded protein. The data points represent the means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.009; **, p 0.007; ***, p 0.002. B and C, <t>SW872</t> cells (treated with vehicle, 500 nM rosiglitazone, and 160 M arachidonic acid) were washed at 37 °C to remove any residual fetal calf serum. The cells were incubated at 37 °C for the indicated times in HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M*. At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble ma- terial was used as a measure of the degraded protein. Trichloroacetic acid-soluble material from untreated cells (vehicle) was compared with the trichloroacetic acid-soluble material from cells treated with 500 nM rosiglitazone (B) or 160 M arachidonic acid (C). The data points rep- resent means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.006; **, p 0.03. DMSO, dimethyl sulfoxide.
Sw 872 Bcrc 60432, supplied by BioResource International Inc, 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/product/sw+872/sw+872+human+liposarcoma+cells+bcrc+60432/pm29389554-81-8-33
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human liposarcoma cell line sw  (Pasteur Institute)
86
Pasteur Institute human liposarcoma cell line sw
FIG. 2. Increased cellular degradation of 125I-2M* by cells treated with PPAR ligands. A, differentiated primary human adi- pocytes were treated with various concentrations of rosiglitazone in the absence of insulin for 24 h prior to the experiment. The cells were then incubated at 37 °C with HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M* with or without RAP (30 g/ml). At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble material was used as a measure of the degraded protein. The data points represent the means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.009; **, p 0.007; ***, p 0.002. B and C, <t>SW872</t> cells (treated with vehicle, 500 nM rosiglitazone, and 160 M arachidonic acid) were washed at 37 °C to remove any residual fetal calf serum. The cells were incubated at 37 °C for the indicated times in HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M*. At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble ma- terial was used as a measure of the degraded protein. Trichloroacetic acid-soluble material from untreated cells (vehicle) was compared with the trichloroacetic acid-soluble material from cells treated with 500 nM rosiglitazone (B) or 160 M arachidonic acid (C). The data points rep- resent means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.006; **, p 0.03. DMSO, dimethyl sulfoxide.
Human Liposarcoma Cell Line Sw, supplied by Pasteur Institute, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/sw+872/872+cell+human+line+liposarcoma+sw/10__31661_slash_gmj__v4i3__335-46-0-10
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sw 872  (AcceGen Biotechnology)
N/A
The SW 872 cell line was initiated by A. Leibovitz in 1974 at the Scott and White Clinic, Temple, Texas from a surgical specimen of a fibrosarcoma removed from a 36 year old male Caucasian.The
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Image Search Results


Figure 2. In vivo application of [D]-K3H3L9. (a) Eighty-six percent remission in s.c. implanted SW-872 liposarcoma xenografts after

Journal: International journal of cancer

Article Title: Oncolytic designer host defense peptide suppresses growth of human liposarcoma.

doi: 10.1002/ijc.25627

Figure Lengend Snippet: Figure 2. In vivo application of [D]-K3H3L9. (a) Eighty-six percent remission in s.c. implanted SW-872 liposarcoma xenografts after

Article Snippet: SW-872 cells were obtained from CLS in 2006 and last authenticated via DNA (STR) profiling by the DSMZ GmbH (DSMZ GmbH, Braunschweig, Germany) in July 2008.

Techniques: In Vivo

Figure 5. Analysis of killing mechanism. (a) TUNEL assay revealed an apoptotic effect of [D]-K3H3L9 toward SW-872 cells and primary

Journal: International journal of cancer

Article Title: Oncolytic designer host defense peptide suppresses growth of human liposarcoma.

doi: 10.1002/ijc.25627

Figure Lengend Snippet: Figure 5. Analysis of killing mechanism. (a) TUNEL assay revealed an apoptotic effect of [D]-K3H3L9 toward SW-872 cells and primary

Article Snippet: SW-872 cells were obtained from CLS in 2006 and last authenticated via DNA (STR) profiling by the DSMZ GmbH (DSMZ GmbH, Braunschweig, Germany) in July 2008.

Techniques: TUNEL Assay

Figure 6. FACS analysis. (a) Distribution of dead SW-872 liposarcoma cells after 2, 15 and 30 min of incubation with a lethal dose of [D]- K3H3L9 (white bar) and PBS (black bar), respectively. FACS analysis revealed a time-dependent rise in the percentage of dead cells. In

Journal: International journal of cancer

Article Title: Oncolytic designer host defense peptide suppresses growth of human liposarcoma.

doi: 10.1002/ijc.25627

Figure Lengend Snippet: Figure 6. FACS analysis. (a) Distribution of dead SW-872 liposarcoma cells after 2, 15 and 30 min of incubation with a lethal dose of [D]- K3H3L9 (white bar) and PBS (black bar), respectively. FACS analysis revealed a time-dependent rise in the percentage of dead cells. In

Article Snippet: SW-872 cells were obtained from CLS in 2006 and last authenticated via DNA (STR) profiling by the DSMZ GmbH (DSMZ GmbH, Braunschweig, Germany) in July 2008.

Techniques: Incubation

FIG. 2. Increased cellular degradation of 125I-2M* by cells treated with PPAR ligands. A, differentiated primary human adi- pocytes were treated with various concentrations of rosiglitazone in the absence of insulin for 24 h prior to the experiment. The cells were then incubated at 37 °C with HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M* with or without RAP (30 g/ml). At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble material was used as a measure of the degraded protein. The data points represent the means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.009; **, p 0.007; ***, p 0.002. B and C, SW872 cells (treated with vehicle, 500 nM rosiglitazone, and 160 M arachidonic acid) were washed at 37 °C to remove any residual fetal calf serum. The cells were incubated at 37 °C for the indicated times in HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M*. At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble ma- terial was used as a measure of the degraded protein. Trichloroacetic acid-soluble material from untreated cells (vehicle) was compared with the trichloroacetic acid-soluble material from cells treated with 500 nM rosiglitazone (B) or 160 M arachidonic acid (C). The data points rep- resent means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.006; **, p 0.03. DMSO, dimethyl sulfoxide.

Journal: Journal of Biological Chemistry

Article Title: Adipocyte Low Density Lipoprotein Receptor-related Protein Gene Expression and Function Is Regulated by Peroxisome Proliferator-activated Receptor γ

doi: 10.1074/jbc.m212989200

Figure Lengend Snippet: FIG. 2. Increased cellular degradation of 125I-2M* by cells treated with PPAR ligands. A, differentiated primary human adi- pocytes were treated with various concentrations of rosiglitazone in the absence of insulin for 24 h prior to the experiment. The cells were then incubated at 37 °C with HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M* with or without RAP (30 g/ml). At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble material was used as a measure of the degraded protein. The data points represent the means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.009; **, p 0.007; ***, p 0.002. B and C, SW872 cells (treated with vehicle, 500 nM rosiglitazone, and 160 M arachidonic acid) were washed at 37 °C to remove any residual fetal calf serum. The cells were incubated at 37 °C for the indicated times in HBSS, 25 mM HEPES, pH 7.45, 10 mg/ml BSA containing 1 mg/ml 125I-2M*. At the beginning of the incubation period and at each time point, the medium was precipitated with ice-cold tricarboxylic acid, and the tricarboxylic acid-soluble ma- terial was used as a measure of the degraded protein. Trichloroacetic acid-soluble material from untreated cells (vehicle) was compared with the trichloroacetic acid-soluble material from cells treated with 500 nM rosiglitazone (B) or 160 M arachidonic acid (C). The data points rep- resent means from triplicate experiments, and the error bars represent the standard error of the mean. *, p 0.006; **, p 0.03. DMSO, dimethyl sulfoxide.

Article Snippet: Cell Culture—The human liposarcoma cell line SW872 (American Type Culture Collection, Manassas, VA) was previously characterized and has been shown to be a good cell model for adipocyte gene expression (42–44).

Techniques: Incubation

FIG. 3. LRP protein levels are increased in SW872 cells upon exposure to PPAR ligands. A, cells were incubated with 1 g/ml fluorescently labeled activated 2M (2M*) (Cy3 monofunctional reac- tive dye) on ice for 45 min in 3:1 Dulbecco’s modified Eagle’s medium/ Ham’s F-12 medium supplement with 2 mg/ml BSA buffered with 10 mM HEPES after an equilibration period of 1 h. The cells were washed with ice-cold PBS three times prior to being fixed with 4% paraformal- dehyde for 10 min at 0 °C. The cells were rinsed with PBS and kept in 2 ml of PBS at room temperature for fluorescence microscopy. The photographs are representative of cells treated with the ligands indi- cated. A 30-fold excess of unlabeled 2M* was used to compete for binding with the fluorescently labeled 2M*. The photographs are normalized so that an increase in intensity on the cell surface and the cell circumference between photographs represents an increase in the fluorescence (i.e. total binding). B, Western blot of cells treated with Me2SO (vehicle control), 160 M arachidonic acid, or 500 nM rosiglita- zone for 24 h. 5 g of total protein was loaded in each lane. The blot was developed using chemiluminescent techniques, and the bands were visualized using the ChemiDoc apparatus. The band at 515 kDa corre- sponds to the -subunit of LRP. C, quantification of Western blot. The 515-kDa band corresponding to LRP was quantified using Quantity One software. All of the samples were normalized to the vehicle-treated cells (control). The Results are shown as the means of triplicate exper- iments, and the error bars represent the standard error of the mean. *, the two-tailed p value is 0.002; **, the two-tailed p value is 0.003.

Journal: Journal of Biological Chemistry

Article Title: Adipocyte Low Density Lipoprotein Receptor-related Protein Gene Expression and Function Is Regulated by Peroxisome Proliferator-activated Receptor γ

doi: 10.1074/jbc.m212989200

Figure Lengend Snippet: FIG. 3. LRP protein levels are increased in SW872 cells upon exposure to PPAR ligands. A, cells were incubated with 1 g/ml fluorescently labeled activated 2M (2M*) (Cy3 monofunctional reac- tive dye) on ice for 45 min in 3:1 Dulbecco’s modified Eagle’s medium/ Ham’s F-12 medium supplement with 2 mg/ml BSA buffered with 10 mM HEPES after an equilibration period of 1 h. The cells were washed with ice-cold PBS three times prior to being fixed with 4% paraformal- dehyde for 10 min at 0 °C. The cells were rinsed with PBS and kept in 2 ml of PBS at room temperature for fluorescence microscopy. The photographs are representative of cells treated with the ligands indi- cated. A 30-fold excess of unlabeled 2M* was used to compete for binding with the fluorescently labeled 2M*. The photographs are normalized so that an increase in intensity on the cell surface and the cell circumference between photographs represents an increase in the fluorescence (i.e. total binding). B, Western blot of cells treated with Me2SO (vehicle control), 160 M arachidonic acid, or 500 nM rosiglita- zone for 24 h. 5 g of total protein was loaded in each lane. The blot was developed using chemiluminescent techniques, and the bands were visualized using the ChemiDoc apparatus. The band at 515 kDa corre- sponds to the -subunit of LRP. C, quantification of Western blot. The 515-kDa band corresponding to LRP was quantified using Quantity One software. All of the samples were normalized to the vehicle-treated cells (control). The Results are shown as the means of triplicate exper- iments, and the error bars represent the standard error of the mean. *, the two-tailed p value is 0.002; **, the two-tailed p value is 0.003.

Article Snippet: Cell Culture—The human liposarcoma cell line SW872 (American Type Culture Collection, Manassas, VA) was previously characterized and has been shown to be a good cell model for adipocyte gene expression (42–44).

Techniques: Incubation, Labeling, Modification, Fluorescence, Microscopy, Binding Assay, Western Blot, Control, Software, Two Tailed Test

FIG. 4. Endogenous LRP mRNA levels in SW872 cells are mod- ulated by PPAR ligands at the transcriptional level. The effect of PPAR ligands on levels of LRP mRNA was determined by incubating SW872 cells for 24 h in Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (3:1) supplemented with complete serum in the presence or absence of the PPAR ligands. A, oleic acid. B, arachidonic acid. C, rosiglitazone. Also a potent inhibitor of RNA polymerase II, -amanitin, was added to SW872 cells in the presence or absence of 500 nM rosigli- tazone to inhibit transcription (C). Reverse transcription was performed on total RNA isolated from these cells, and multiplex PCR was per- formed using LRP and 18 S gene-specific primers (relative quantitative RT-PCR). PCR products were visualized by EtBr staining on the Chemi- Doc, and the band intensities were determined using Quantity One software. The intensity of the LRP product was divided by the intensity of the 18 S product to obtain a value termed relative intensity. These results were normalized to the control and shown as fold increases. The bars in each graph represent the means of triplicate experiments, and the error bars represent the standard error of the mean. One-tailed p values from paired t test are shown above the bars to indicate signifi- cant differences.

Journal: Journal of Biological Chemistry

Article Title: Adipocyte Low Density Lipoprotein Receptor-related Protein Gene Expression and Function Is Regulated by Peroxisome Proliferator-activated Receptor γ

doi: 10.1074/jbc.m212989200

Figure Lengend Snippet: FIG. 4. Endogenous LRP mRNA levels in SW872 cells are mod- ulated by PPAR ligands at the transcriptional level. The effect of PPAR ligands on levels of LRP mRNA was determined by incubating SW872 cells for 24 h in Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (3:1) supplemented with complete serum in the presence or absence of the PPAR ligands. A, oleic acid. B, arachidonic acid. C, rosiglitazone. Also a potent inhibitor of RNA polymerase II, -amanitin, was added to SW872 cells in the presence or absence of 500 nM rosigli- tazone to inhibit transcription (C). Reverse transcription was performed on total RNA isolated from these cells, and multiplex PCR was per- formed using LRP and 18 S gene-specific primers (relative quantitative RT-PCR). PCR products were visualized by EtBr staining on the Chemi- Doc, and the band intensities were determined using Quantity One software. The intensity of the LRP product was divided by the intensity of the 18 S product to obtain a value termed relative intensity. These results were normalized to the control and shown as fold increases. The bars in each graph represent the means of triplicate experiments, and the error bars represent the standard error of the mean. One-tailed p values from paired t test are shown above the bars to indicate signifi- cant differences.

Article Snippet: Cell Culture—The human liposarcoma cell line SW872 (American Type Culture Collection, Manassas, VA) was previously characterized and has been shown to be a good cell model for adipocyte gene expression (42–44).

Techniques: Modification, Reverse Transcription, Isolation, Multiplex Assay, Quantitative RT-PCR, Staining, Software, Control, One-tailed Test

FIG. 6. PPAR-RXR heterodimers selectively bind PPRE in the LRP promoter. EMSA were performed on oligomers correspond- ing the PPREs of LRP, hACOX, and LRPmut. A, the PPREs of the human LRP and ACOX genes are shown compared with the consensus PPRE along with the sequence of the mutant LRP PPRE. The mis- matches for each PPRE are underlined. B, oligomers were incubated on ice for 20 min with various components. Unprogrammed reticulocyte lysate (TNT reaction containing empty pSPORT1 vector) was used as a control (lanes 1, 7, and 13). TNT in vitro transcribed/translated PPAR and RXR were incubated with radiolabeled PPRE oligomers (lanes 2, 8, and 14). To control for specificity of binding competition experiments were performed by adding increasing amounts of unlabeled oligomer to the binding reactions. LRP binding reactions were competed using cold hACOX oligomers at 5, 10, 25, and 50 excess (lanes 3–6), and the hACOX binding reactions were competed using cold LRP oligomers at 5, 10, 25, and 50 excess (lanes 9–12). C, nuclear extracts from primary human adipocytes were incubated with radiolabeled oligomer (lanes 1, 6, and 11) on ice for 20 min. Specificity of binding was again tested using unlabeled oligomers in competition experiments as de- scribed above (lanes 2–5 and 7–10). D, gel supershift analysis was used to determine whether PPAR was present in shift seen with nuclear extracts. Free probe (first lane) was used as a control reaction. The nuclear extracts (18 g) were incubated with labeled oligomer and run in the second lane. NuShift anti-PPAR antibody (4 l) was incubated with 18 g of nuclear extracts from SW872 cells prior to being added to the EMSA reaction mix containing labeled oligomers (third lane). A nonspecific antibody (mouse monoclonal anti--actin) was used as a negative control for the supershift (fourth lane). The arrows indicate the shift and the supershift.

Journal: Journal of Biological Chemistry

Article Title: Adipocyte Low Density Lipoprotein Receptor-related Protein Gene Expression and Function Is Regulated by Peroxisome Proliferator-activated Receptor γ

doi: 10.1074/jbc.m212989200

Figure Lengend Snippet: FIG. 6. PPAR-RXR heterodimers selectively bind PPRE in the LRP promoter. EMSA were performed on oligomers correspond- ing the PPREs of LRP, hACOX, and LRPmut. A, the PPREs of the human LRP and ACOX genes are shown compared with the consensus PPRE along with the sequence of the mutant LRP PPRE. The mis- matches for each PPRE are underlined. B, oligomers were incubated on ice for 20 min with various components. Unprogrammed reticulocyte lysate (TNT reaction containing empty pSPORT1 vector) was used as a control (lanes 1, 7, and 13). TNT in vitro transcribed/translated PPAR and RXR were incubated with radiolabeled PPRE oligomers (lanes 2, 8, and 14). To control for specificity of binding competition experiments were performed by adding increasing amounts of unlabeled oligomer to the binding reactions. LRP binding reactions were competed using cold hACOX oligomers at 5, 10, 25, and 50 excess (lanes 3–6), and the hACOX binding reactions were competed using cold LRP oligomers at 5, 10, 25, and 50 excess (lanes 9–12). C, nuclear extracts from primary human adipocytes were incubated with radiolabeled oligomer (lanes 1, 6, and 11) on ice for 20 min. Specificity of binding was again tested using unlabeled oligomers in competition experiments as de- scribed above (lanes 2–5 and 7–10). D, gel supershift analysis was used to determine whether PPAR was present in shift seen with nuclear extracts. Free probe (first lane) was used as a control reaction. The nuclear extracts (18 g) were incubated with labeled oligomer and run in the second lane. NuShift anti-PPAR antibody (4 l) was incubated with 18 g of nuclear extracts from SW872 cells prior to being added to the EMSA reaction mix containing labeled oligomers (third lane). A nonspecific antibody (mouse monoclonal anti--actin) was used as a negative control for the supershift (fourth lane). The arrows indicate the shift and the supershift.

Article Snippet: Cell Culture—The human liposarcoma cell line SW872 (American Type Culture Collection, Manassas, VA) was previously characterized and has been shown to be a good cell model for adipocyte gene expression (42–44).

Techniques: Sequencing, Mutagenesis, Incubation, Plasmid Preparation, Control, In Vitro, Binding Assay, Labeling, Negative Control

FIG. 7. LRP promoter confers regulation of luciferase activity by rosiglitazone. A, the promoter region of human LRP (0 to 1200) was cloned into pGL3-Basic. This construct was then transiently co- transfected into SW872 cells along with the Renilla luciferase reporter vector pRL-CMV using the calcium-phosphate precipitation method. Luciferase activities for each of firefly and Renilla luciferase were determined using the dual luciferase assay system. The intensity of firefly luciferase is shown as a function of Renilla luciferase (relative activity). The relative intensity is normalized to the control (vehicle treatment) values. The means of triplicate experiments are graphed, and the error bars represent the standard error of the mean. The one-tailed p values from paired t tests are shown for significant differ- ences. B, pGL3-LRP was subjected to site-directed mutagenesis, in which the half-site of the DR-1 to which PPAR binds, was mutated so that 5 of 6 nucleotides no longer matched the consensus. PPREs of pGL3-LRP and pGL3-LRPmutant PPRE are shown with the consensus sequence for comparison. Point mutations are underlined, and the mismatches present in endogenous PPRE are in italics. C, pGL3- LRPmutant PPRE, was transiently co-transfected with pRL-CMV into SW872 cells, and their respective luciferase activities were determined using the dual luciferase assay system. The relative intensity was calculated, and the means of triplicate experiments are shown. The values of relative intensity are normalized to the control (vehicle-treat- ed) cells. The error bars represent the standard error of the mean. Rosiglitazone has a significant effect on pGL3-LRP; however, it is not significant for pGL3-LRPmutant PPRE. The one-tailed p values from t tests were 0.02 and 0.2, respectively.

Journal: Journal of Biological Chemistry

Article Title: Adipocyte Low Density Lipoprotein Receptor-related Protein Gene Expression and Function Is Regulated by Peroxisome Proliferator-activated Receptor γ

doi: 10.1074/jbc.m212989200

Figure Lengend Snippet: FIG. 7. LRP promoter confers regulation of luciferase activity by rosiglitazone. A, the promoter region of human LRP (0 to 1200) was cloned into pGL3-Basic. This construct was then transiently co- transfected into SW872 cells along with the Renilla luciferase reporter vector pRL-CMV using the calcium-phosphate precipitation method. Luciferase activities for each of firefly and Renilla luciferase were determined using the dual luciferase assay system. The intensity of firefly luciferase is shown as a function of Renilla luciferase (relative activity). The relative intensity is normalized to the control (vehicle treatment) values. The means of triplicate experiments are graphed, and the error bars represent the standard error of the mean. The one-tailed p values from paired t tests are shown for significant differ- ences. B, pGL3-LRP was subjected to site-directed mutagenesis, in which the half-site of the DR-1 to which PPAR binds, was mutated so that 5 of 6 nucleotides no longer matched the consensus. PPREs of pGL3-LRP and pGL3-LRPmutant PPRE are shown with the consensus sequence for comparison. Point mutations are underlined, and the mismatches present in endogenous PPRE are in italics. C, pGL3- LRPmutant PPRE, was transiently co-transfected with pRL-CMV into SW872 cells, and their respective luciferase activities were determined using the dual luciferase assay system. The relative intensity was calculated, and the means of triplicate experiments are shown. The values of relative intensity are normalized to the control (vehicle-treat- ed) cells. The error bars represent the standard error of the mean. Rosiglitazone has a significant effect on pGL3-LRP; however, it is not significant for pGL3-LRPmutant PPRE. The one-tailed p values from t tests were 0.02 and 0.2, respectively.

Article Snippet: Cell Culture—The human liposarcoma cell line SW872 (American Type Culture Collection, Manassas, VA) was previously characterized and has been shown to be a good cell model for adipocyte gene expression (42–44).

Techniques: Luciferase, Activity Assay, Clone Assay, Construct, Transfection, Plasmid Preparation, Control, One-tailed Test, Mutagenesis, Sequencing, Comparison