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  • 99
    Thermo Fisher fugene 6
    Fugene 6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 766 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega fugene 6
    Fugene 6, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 11560 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6/product/Promega
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    92
    Boehringer Mannheim fugene 6
    Glomerular H-E staining scores in the kidney samples on day 4 (4-d) and weeks 3 (3-w) and 5 (5-w) after repeated injections of <t>pCAG/DT-A/FuGENE™6</t> complex, pCAG/FuGENE™6 complex or mock injections [FuGENE™6 only or PBS(-) only]. At least 100 glomeruli in the H-E-stained sections were randomly chosen per kidney and the lesions were quantified. A total of 4 kidneys for each group were inspected. The glomerular lesions were graded from 0 to 2: score 0, normal; score 1, weak to mild change (exhibiting mild mesangial cell proliferation and mesangial matrix expansion); score 2, severe change (exhibiting lobular structure of glomeruli and adhesion of glomeruli to the inner surface of Bowman's capsule).
    Fugene 6, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 955 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6/product/Boehringer Mannheim
    Average 92 stars, based on 955 article reviews
    Price from $9.99 to $1999.99
    fugene 6 - by Bioz Stars, 2020-10
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    92
    Boehringer Mannheim fugene 6 transfection reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene 6 Transfection Reagent, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 558 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6 transfection reagent/product/Boehringer Mannheim
    Average 92 stars, based on 558 article reviews
    Price from $9.99 to $1999.99
    fugene 6 transfection reagent - by Bioz Stars, 2020-10
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    99
    Thermo Fisher fugene6
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene6, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 505 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene6/product/Thermo Fisher
    Average 99 stars, based on 505 article reviews
    Price from $9.99 to $1999.99
    fugene6 - by Bioz Stars, 2020-10
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    92
    Boehringer Mannheim fugene 6 reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene 6 Reagent, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6 reagent/product/Boehringer Mannheim
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    Price from $9.99 to $1999.99
    fugene 6 reagent - by Bioz Stars, 2020-10
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    94
    Thermo Fisher fugene 6 transfection reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene 6 Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6 transfection reagent/product/Thermo Fisher
    Average 94 stars, based on 110 article reviews
    Price from $9.99 to $1999.99
    fugene 6 transfection reagent - by Bioz Stars, 2020-10
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    99
    Thermo Fisher fugene 6 reagent
    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by <t>FuGENE</t> 6 <t>Transfection</t> reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.
    Fugene 6 Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 132 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fugene 6 reagent/product/Thermo Fisher
    Average 99 stars, based on 132 article reviews
    Price from $9.99 to $1999.99
    fugene 6 reagent - by Bioz Stars, 2020-10
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    Image Search Results


    Glomerular H-E staining scores in the kidney samples on day 4 (4-d) and weeks 3 (3-w) and 5 (5-w) after repeated injections of pCAG/DT-A/FuGENE™6 complex, pCAG/FuGENE™6 complex or mock injections [FuGENE™6 only or PBS(-) only]. At least 100 glomeruli in the H-E-stained sections were randomly chosen per kidney and the lesions were quantified. A total of 4 kidneys for each group were inspected. The glomerular lesions were graded from 0 to 2: score 0, normal; score 1, weak to mild change (exhibiting mild mesangial cell proliferation and mesangial matrix expansion); score 2, severe change (exhibiting lobular structure of glomeruli and adhesion of glomeruli to the inner surface of Bowman's capsule).

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: Glomerular H-E staining scores in the kidney samples on day 4 (4-d) and weeks 3 (3-w) and 5 (5-w) after repeated injections of pCAG/DT-A/FuGENE™6 complex, pCAG/FuGENE™6 complex or mock injections [FuGENE™6 only or PBS(-) only]. At least 100 glomeruli in the H-E-stained sections were randomly chosen per kidney and the lesions were quantified. A total of 4 kidneys for each group were inspected. The glomerular lesions were graded from 0 to 2: score 0, normal; score 1, weak to mild change (exhibiting mild mesangial cell proliferation and mesangial matrix expansion); score 2, severe change (exhibiting lobular structure of glomeruli and adhesion of glomeruli to the inner surface of Bowman's capsule).

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Staining

    Serum levels of BUN, CRT, Na + , K + and Cl - in the mice repeatedly injected with pCAG/DT-A/FuGENE™6 complex, pCAG/FuGENE™6 complex, FuGENE™6 only or PBS(-) only. Serum sampling was performed on day 4(4-d) and weeks 3 (3-w) and 5 (5-w).

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: Serum levels of BUN, CRT, Na + , K + and Cl - in the mice repeatedly injected with pCAG/DT-A/FuGENE™6 complex, pCAG/FuGENE™6 complex, FuGENE™6 only or PBS(-) only. Serum sampling was performed on day 4(4-d) and weeks 3 (3-w) and 5 (5-w).

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Mouse Assay, Injection, Sampling

    ( A ) Detection of exogenous pCAG/DT-A by PCR analysis. Genomic DNA was extracted from organs of the mice injected with pCAG/DT-A/FuGENE™6 complex or PBS(-) (mock injection). Detection of pCAG/DT-A was performed by PCR using DTA-S/DTA-2RV primer set (see Figure 1A ). This PCR yielded a product of 267 bp [indicated by an arrow in ( A )]. C indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to PCR as a negative control. PC indicates that pCAG/DT-A (4 ng) was subjected to PCR as a positive control. ( B ) Detection of DT-A mRNA by RT-PCR analysis. Detection of DT-A mRNA was performed by RT-PCR using βA-1/DTA-RV primer set (see Figure 1A ), which yielded an expected band of 300 bp [indicated by an arrow in ( B )]. In the lower panel of ( B ), the expression pattern of endogenous β-actin mRNA is indicated. Lu, lung; Br, brain; H, heart; K, kidney; In, intestine; L, liver. C-1 indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to PCR. C-2 indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to reverse transcription and the resulting solution was then PCR-amplified. PC indicates that pCAG/DT-A (4 ng) was subjected to RT-PCR as a positive control. A few non-specifically amplified bands above the 300-bp band were observed in lane C-2, and approximately 600-bp bands [indicated by an arrowhead in ( B )], probably corresponding to the products obtained through amplification of pCAG/DT-A, were also seen in lane PC.

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: ( A ) Detection of exogenous pCAG/DT-A by PCR analysis. Genomic DNA was extracted from organs of the mice injected with pCAG/DT-A/FuGENE™6 complex or PBS(-) (mock injection). Detection of pCAG/DT-A was performed by PCR using DTA-S/DTA-2RV primer set (see Figure 1A ). This PCR yielded a product of 267 bp [indicated by an arrow in ( A )]. C indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to PCR as a negative control. PC indicates that pCAG/DT-A (4 ng) was subjected to PCR as a positive control. ( B ) Detection of DT-A mRNA by RT-PCR analysis. Detection of DT-A mRNA was performed by RT-PCR using βA-1/DTA-RV primer set (see Figure 1A ), which yielded an expected band of 300 bp [indicated by an arrow in ( B )]. In the lower panel of ( B ), the expression pattern of endogenous β-actin mRNA is indicated. Lu, lung; Br, brain; H, heart; K, kidney; In, intestine; L, liver. C-1 indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to PCR. C-2 indicates that tail DNA (1 μg) from a B6C3F1 mouse was directly subjected to reverse transcription and the resulting solution was then PCR-amplified. PC indicates that pCAG/DT-A (4 ng) was subjected to RT-PCR as a positive control. A few non-specifically amplified bands above the 300-bp band were observed in lane C-2, and approximately 600-bp bands [indicated by an arrowhead in ( B )], probably corresponding to the products obtained through amplification of pCAG/DT-A, were also seen in lane PC.

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Polymerase Chain Reaction, Mouse Assay, Injection, Negative Control, Positive Control, Reverse Transcription Polymerase Chain Reaction, Expressing, Amplification

    EGFP fluorescence in 4% paraformaldehyde-fixed heart (a), lung (b,c) and kidneys (d-f) of mice injected 6 times with pCE-29/ FuGENE™6 complex. (a) Inner surface of the heart cut into slices. Note that the luminal surface of the heart exhibited focal and slight EGFP fluorescence (indicated by arrows). (b,c) Inner surface (b) of the lung cut into slices and outer (c) surface of the lung. Note focal or patchy expression of EGFP fluorescence in each sample (indicated by arrows). (d) Inner surface of the kidney cut into slices. Note that several glomeruli expressed EGFP fluorescence (indicated by arrows). (e) A cryostat section of the kidney in (d). Patchy expression for EGFP fluorescence (indicated by an arrow) in glomeruli was noted. Also, some renal tubules exhibited faint fluorescence (indicated by arrowheads). (f) A figure magnified from the boxed region in (e). EGFP fluorescence was patchy in the glomerulus (indicated by arrows).

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: EGFP fluorescence in 4% paraformaldehyde-fixed heart (a), lung (b,c) and kidneys (d-f) of mice injected 6 times with pCE-29/ FuGENE™6 complex. (a) Inner surface of the heart cut into slices. Note that the luminal surface of the heart exhibited focal and slight EGFP fluorescence (indicated by arrows). (b,c) Inner surface (b) of the lung cut into slices and outer (c) surface of the lung. Note focal or patchy expression of EGFP fluorescence in each sample (indicated by arrows). (d) Inner surface of the kidney cut into slices. Note that several glomeruli expressed EGFP fluorescence (indicated by arrows). (e) A cryostat section of the kidney in (d). Patchy expression for EGFP fluorescence (indicated by an arrow) in glomeruli was noted. Also, some renal tubules exhibited faint fluorescence (indicated by arrowheads). (f) A figure magnified from the boxed region in (e). EGFP fluorescence was patchy in the glomerulus (indicated by arrows).

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Fluorescence, Mouse Assay, Injection, Expressing

    H-E staining of glomeruli and renal tubules in the kidneys of mice injected 6 times with pCAG/DT-A/FuGENE™6 complex (a-i), pCAG/FuGENE™6 complex (j) or PBS(-) only (k) on day 4 (a-g) and weeks 3 (h) and 5 (i). On day 4, proliferation of mesangial cells, which causes loss of Bowman's space [indicated by arrows in (a) and (b)] and attenuation of glomerular capillaries [indicated by arrowheads in (c)] were remarkable. In some cases, findings of cellular degeneration [indicated by arrows in (c) and (e)] and appearance of lobular segmental structure of glomerulus (d-f) were observed. Infiltration of lymphocytes into interstitum and renal tubules [indicated by arrowheads in (g)] was sometimes observed. However, no gross alteration in glomeruli was noted in the groups with injection of pCAG/FuGENE™6 complex (j) and of mock injections with FuGENE™6 only (data not shown) or PBS(-) only (k). On week 3, slight recovery was observed in the mice treated with pCAG/DT-A/FuGENE™6 complex, although debris [indicated by an arrow in (h)] existed in Bowman's space. By week 5, the morphology of glomeruli had almost completely recovered, although proteinaceous substances were often observed in Bowman's space (i). (l), Glomerulus of intact kidney.

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: H-E staining of glomeruli and renal tubules in the kidneys of mice injected 6 times with pCAG/DT-A/FuGENE™6 complex (a-i), pCAG/FuGENE™6 complex (j) or PBS(-) only (k) on day 4 (a-g) and weeks 3 (h) and 5 (i). On day 4, proliferation of mesangial cells, which causes loss of Bowman's space [indicated by arrows in (a) and (b)] and attenuation of glomerular capillaries [indicated by arrowheads in (c)] were remarkable. In some cases, findings of cellular degeneration [indicated by arrows in (c) and (e)] and appearance of lobular segmental structure of glomerulus (d-f) were observed. Infiltration of lymphocytes into interstitum and renal tubules [indicated by arrowheads in (g)] was sometimes observed. However, no gross alteration in glomeruli was noted in the groups with injection of pCAG/FuGENE™6 complex (j) and of mock injections with FuGENE™6 only (data not shown) or PBS(-) only (k). On week 3, slight recovery was observed in the mice treated with pCAG/DT-A/FuGENE™6 complex, although debris [indicated by an arrow in (h)] existed in Bowman's space. By week 5, the morphology of glomeruli had almost completely recovered, although proteinaceous substances were often observed in Bowman's space (i). (l), Glomerulus of intact kidney.

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Staining, Mouse Assay, Injection

    Electron microscopic analysis of affected glomeruli (a, c-f, h), and tubules (g) on day 4 (a, c-g) and week 5 (h) after injection of pCAG/DT-A/FuGENE™6 complex. In the affected glomeruli (a), severe matrix expansion with numerous electron-dense deposits was detected in the mesangial matrix. The capillary lumen of endothelial cells was deformed, probably due to proliferation of active mesangial cells. Degeneration of mesangial cells was sometimes observed [indicated by an arrow in (a)]. In contrast, the control glomeruli derived from mock injection of PBS(-) on day 4 (b) exhibited normal appearance. Notably, formation of focal deposits in glomerular basement membrane was often remarkable [indicated by arrows in (c-f)]. The initial, small deposits of glomerular basement membrane are shown by arrowheads in (d) and (e). Incorporation of lipids, probably DNA/lipid complexes that had been re-uptaken by renal tubules, was noted in the cytoplasm of renal tubules [indicated by arrows in (g)]. On week 5, nearly normal glomerular structure was observed (h).

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: Electron microscopic analysis of affected glomeruli (a, c-f, h), and tubules (g) on day 4 (a, c-g) and week 5 (h) after injection of pCAG/DT-A/FuGENE™6 complex. In the affected glomeruli (a), severe matrix expansion with numerous electron-dense deposits was detected in the mesangial matrix. The capillary lumen of endothelial cells was deformed, probably due to proliferation of active mesangial cells. Degeneration of mesangial cells was sometimes observed [indicated by an arrow in (a)]. In contrast, the control glomeruli derived from mock injection of PBS(-) on day 4 (b) exhibited normal appearance. Notably, formation of focal deposits in glomerular basement membrane was often remarkable [indicated by arrows in (c-f)]. The initial, small deposits of glomerular basement membrane are shown by arrowheads in (d) and (e). Incorporation of lipids, probably DNA/lipid complexes that had been re-uptaken by renal tubules, was noted in the cytoplasm of renal tubules [indicated by arrows in (g)]. On week 5, nearly normal glomerular structure was observed (h).

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Injection, Derivative Assay

    PAS and PAM staining of kidney sections sampled day 4 (d-f) and weeks 3 (g-i) and 5 (j-l) after intravenous administration of pCAG/DT-A/FuGENE™6 complex. Note that increased deposition of mesangial matrix, as revealed by increased staining by PAM, was evident in the glomerulus on day 4 (f), but that in later stages (weeks 3 and 5) such deposits were decreased (i,l). (a-c) indicate intact kidney.

    Journal: BMC Nephrology

    Article Title: A new mouse model for renal lesions produced by intravenous injection of diphtheria toxin A-chain expression plasmid

    doi: 10.1186/1471-2369-5-4

    Figure Lengend Snippet: PAS and PAM staining of kidney sections sampled day 4 (d-f) and weeks 3 (g-i) and 5 (j-l) after intravenous administration of pCAG/DT-A/FuGENE™6 complex. Note that increased deposition of mesangial matrix, as revealed by increased staining by PAM, was evident in the glomerulus on day 4 (f), but that in later stages (weeks 3 and 5) such deposits were decreased (i,l). (a-c) indicate intact kidney.

    Article Snippet: Forty μl of FuGENE™6 (Boehringer Mannheim GmbH, Mannheim, Germany), a transfection reagent (lipid), was diluted with 60 μl of PBS(-), and then added to 20 μg of circular plasmid DNA dissolved in 100 μl of PBS(-) according to the manufacturer's protocol.

    Techniques: Staining

    Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by FuGENE 6 Transfection reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Effects of mutant p53 expression on human 15-lipoxygenase-promoter activity and murine 12/15-lipoxygenase gene expression: Evidence that 15-lipoxygenase is a mutator gene

    doi:

    Figure Lengend Snippet: Schematic linear map of human 15-LO )1val cells. Plasmids containing various lengths of human 15-LO promoter 5′ of the luciferase gene were constructed as described in Materials and Methods . The sequence numbers in the map correspond with those published by Kelavkar et al. )1 cells by FuGENE 6 Transfection reagent. The cells were grown in appropriate medium containing 10% FBS at 37, 32, and 39°C, respectively, for 24 h, depending on individual experiments, before being harvested for determination of luciferase activity. Each experiment was repeated at least three times with duplicate samples. The values shown on the right are the averages ±SD of the relative promoter activities.

    Article Snippet: After 24 h, the plasmid DNA (1 μg) was transfected into the cells by FuGENE 6 Transfection reagent (Boehringer Mannheim).

    Techniques: Luciferase, Construct, Sequencing, Transfection, Activity Assay

    Immunocytofluorescence detection of receptors in DU145 cells. DU145 cells were seeded on two-well Lab Tek Chamber slides (Nalge) 18 hr before transfection. One to two micrograms of DNA per 10 5 cells was transfected either with the AR (unliganded or liganded), TR4, or ER alone or in combination with the FuGENE6 transfection reagent (Boehringer-Mannheim). After 24 hr transfection, cells were treated with 100 nM DHT or ethanol. Immunostaining was performed by incubation with the anti-AR polyclonal antibody, anti-TR4 mAb, or anti-ERα mAb, followed by incubation with either fluorescein-conjugated goat anti-rabbit or anti-mouse antibodies (ICN). The red signal represents the AR, the green signal represents the TR4 or ER, and the yellow signal represents colocalization of the two signals in cotransfection of both receptors (the AR with TR4 or ER). Shown is the immunostaining of a single transfection of ( A ) unliganded AR, ( B ) 100 nM DHT treatment of the AR, ( C ) TR4, and ( D ) ER, and for cotransfection of ( E ) AR with TR4 and ( F ) AR with ER. The slides were photographed under 100-fold magnification by using confocal microscopy.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Convergence of two repressors through heterodimer formation of androgen receptor and testicular orphan receptor-4: A unique signaling pathway in the steroid receptor superfamily

    doi:

    Figure Lengend Snippet: Immunocytofluorescence detection of receptors in DU145 cells. DU145 cells were seeded on two-well Lab Tek Chamber slides (Nalge) 18 hr before transfection. One to two micrograms of DNA per 10 5 cells was transfected either with the AR (unliganded or liganded), TR4, or ER alone or in combination with the FuGENE6 transfection reagent (Boehringer-Mannheim). After 24 hr transfection, cells were treated with 100 nM DHT or ethanol. Immunostaining was performed by incubation with the anti-AR polyclonal antibody, anti-TR4 mAb, or anti-ERα mAb, followed by incubation with either fluorescein-conjugated goat anti-rabbit or anti-mouse antibodies (ICN). The red signal represents the AR, the green signal represents the TR4 or ER, and the yellow signal represents colocalization of the two signals in cotransfection of both receptors (the AR with TR4 or ER). Shown is the immunostaining of a single transfection of ( A ) unliganded AR, ( B ) 100 nM DHT treatment of the AR, ( C ) TR4, and ( D ) ER, and for cotransfection of ( E ) AR with TR4 and ( F ) AR with ER. The slides were photographed under 100-fold magnification by using confocal microscopy.

    Article Snippet: One or two micrograms of DNA per 105 cells was transfected by the FuGENE6 transfection reagent (Boehringer-Mannheim).

    Techniques: Transfection, Immunostaining, Incubation, Cotransfection, Confocal Microscopy