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  • 99
    Promega fugene6
    Analysis of cellular protein expression in ZIKV-infected hBMECs. (A to C) hBMECs were mock infected or infected with ZIKV, and 1 to 9 dpi supernatants were analyzed in an ELISA (R D Systems) for CCL/RANTES (A), IFN-β (B), and IFN-λ (C) levels relative to antigen standards. As an hBMEC IFN-β response control, we transfected hBMECs with poly(I/C) (1 µg/ml) and <t>Fugene6</t> at 3:1 and evaluated secreted IFN-β levels in supernatants via ELISA (36 h posttransfection) (D) Western blot analysis of MXA and IFIT1 genes, and GAPDH controls, in lysates from mock-infected or ZIKV-infected hBMECs (1 to 9 dpi). IFIT and MxA protein levels in ZIKV-infected hBMECs 9 dpi versus results 6 h post-IFN-α treatment (1,000 U/ml).
    Fugene6, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 5621 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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
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    92
    Fisher Scientific 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 Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Roche fugene 6
    Effect of cytofectins on CT DNA uptake by macrophages. J774 cells were incubated for up to 48 hr with 0·4–10 µg/ml EMA-CT DNA with or without Fugene 6. DNA uptake was assessed by flow cytometric analysis of the cells collected at different time-points. The levels of DNA uptake are represented as mean fluorescence intensity (MFI) for each group. There was a dose- and time-dependent increase of DNA uptake in the presence of Fugene 6.
    Fugene 6, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 41961 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher fugene
    Effect of cytofectins on CT DNA uptake by macrophages. J774 cells were incubated for up to 48 hr with 0·4–10 µg/ml EMA-CT DNA with or without Fugene 6. DNA uptake was assessed by flow cytometric analysis of the cells collected at different time-points. The levels of DNA uptake are represented as mean fluorescence intensity (MFI) for each group. There was a dose- and time-dependent increase of DNA uptake in the presence of Fugene 6.
    Fugene, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 395 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Analysis of cellular protein expression in ZIKV-infected hBMECs. (A to C) hBMECs were mock infected or infected with ZIKV, and 1 to 9 dpi supernatants were analyzed in an ELISA (R D Systems) for CCL/RANTES (A), IFN-β (B), and IFN-λ (C) levels relative to antigen standards. As an hBMEC IFN-β response control, we transfected hBMECs with poly(I/C) (1 µg/ml) and Fugene6 at 3:1 and evaluated secreted IFN-β levels in supernatants via ELISA (36 h posttransfection) (D) Western blot analysis of MXA and IFIT1 genes, and GAPDH controls, in lysates from mock-infected or ZIKV-infected hBMECs (1 to 9 dpi). IFIT and MxA protein levels in ZIKV-infected hBMECs 9 dpi versus results 6 h post-IFN-α treatment (1,000 U/ml).

    Journal: mBio

    Article Title: Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells

    doi: 10.1128/mBio.00952-17

    Figure Lengend Snippet: Analysis of cellular protein expression in ZIKV-infected hBMECs. (A to C) hBMECs were mock infected or infected with ZIKV, and 1 to 9 dpi supernatants were analyzed in an ELISA (R D Systems) for CCL/RANTES (A), IFN-β (B), and IFN-λ (C) levels relative to antigen standards. As an hBMEC IFN-β response control, we transfected hBMECs with poly(I/C) (1 µg/ml) and Fugene6 at 3:1 and evaluated secreted IFN-β levels in supernatants via ELISA (36 h posttransfection) (D) Western blot analysis of MXA and IFIT1 genes, and GAPDH controls, in lysates from mock-infected or ZIKV-infected hBMECs (1 to 9 dpi). IFIT and MxA protein levels in ZIKV-infected hBMECs 9 dpi versus results 6 h post-IFN-α treatment (1,000 U/ml).

    Article Snippet: Levels of IFN-β, IFN-λ, and CCL5/RANTES in the supernatants of mock- and ZIKV-infected hBMECs at 12 hpi to 9 dpi were measured using a DuoSet ELISA (R & D Systems). hBMECs transfected with poly(I/C) (1 µg/ml) and Fugene6 (3:1; Promega) were similarly evaluated for secreted IFN-β at 36 h posttransfection.

    Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Transfection, Western Blot

    ChIP analysis indicating that ZPO2 and ZBTB32 occupy the Gata3 promoter. EpH4.9 cells were grown in 150-mm culture dishes and cotransfected with full-length Zpo2 and Myc-tagged ZBTB32 (generous gifts from Dr. I.-Cheng Ho, Harvard Medical School) constructs via FuGENE6 transfection reagent (Promega). ChIP analysis was performed using the ChIP-IT High-Sensitivity Kit (Active Motif) in accordance with the manufacturer’s protocol. qPCR for ChIP samples was performed using Gata3 promoter sequence primers listed in the text. ChIP analysis indicated that both ZPO2 and ZBTB32 occupy the promoter sequence of GATA3.

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

    Article Title: ZNF503/Zpo2 drives aggressive breast cancer progression by down-regulation of GATA3 expression

    doi: 10.1073/pnas.1701690114

    Figure Lengend Snippet: ChIP analysis indicating that ZPO2 and ZBTB32 occupy the Gata3 promoter. EpH4.9 cells were grown in 150-mm culture dishes and cotransfected with full-length Zpo2 and Myc-tagged ZBTB32 (generous gifts from Dr. I.-Cheng Ho, Harvard Medical School) constructs via FuGENE6 transfection reagent (Promega). ChIP analysis was performed using the ChIP-IT High-Sensitivity Kit (Active Motif) in accordance with the manufacturer’s protocol. qPCR for ChIP samples was performed using Gata3 promoter sequence primers listed in the text. ChIP analysis indicated that both ZPO2 and ZBTB32 occupy the promoter sequence of GATA3.

    Article Snippet: EpH4.9 cells were cultured in 10-cm culture dishes and cotransfected with 7 µg of V5-tagged full-length Zpo2 and 7 µg of Myc-tagged ZBTB32 using FuGENE6 transfection reagent (Promega).

    Techniques: Chromatin Immunoprecipitation, Construct, Transfection, Real-time Polymerase Chain Reaction, Sequencing

    Z potential of control microparticles and of microparticles treated with PEI, LF2000 and FuGENE 6 ® . Note: The Zeta potential of the functionalized microparticle is also shown. Abbreviations: FMP, functionalized microparticle; PEI, polyethyleneimine; LF, Lipofectamine™.

    Journal: International Journal of Nanomedicine

    Article Title: Enhancing microparticle internalization by nonphagocytic cells through the use of noncovalently conjugated polyethyleneimine

    doi: 10.2147/IJN.S34635

    Figure Lengend Snippet: Z potential of control microparticles and of microparticles treated with PEI, LF2000 and FuGENE 6 ® . Note: The Zeta potential of the functionalized microparticle is also shown. Abbreviations: FMP, functionalized microparticle; PEI, polyethyleneimine; LF, Lipofectamine™.

    Article Snippet: PEI, LF2000, and FuGENE 6 cytotoxicity assay The cytotoxicity of PEI 25 KDa (Sigma-Aldrich, St Louis, MO) used at three different concentrations (0.05, 0.10, and 0.15 mM), of LF2000 (Life Technologies), and of FuGENE 6 (Promega Corporation, Fitchburg, WI), used at the concentrations recommended by the manufacturers, was evaluated after the transfection procedure by assessing two different parameters: the percentage of cells that remained attached to the dish (normalized to the control group), and the viability of the attached cells.

    Techniques:

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    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: For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS(-) (without DNA) or PBS(-) only was prepared.

    Techniques: Staining

    Effect of cytofectins on CT DNA uptake by macrophages. J774 cells were incubated for up to 48 hr with 0·4–10 µg/ml EMA-CT DNA with or without Fugene 6. DNA uptake was assessed by flow cytometric analysis of the cells collected at different time-points. The levels of DNA uptake are represented as mean fluorescence intensity (MFI) for each group. There was a dose- and time-dependent increase of DNA uptake in the presence of Fugene 6.

    Journal: Immunology

    Article Title: Effect of cytofectins on the immune response of murine macrophages to mammalian DNA

    doi: 10.1046/j.1365-2567.2003.01653.x

    Figure Lengend Snippet: Effect of cytofectins on CT DNA uptake by macrophages. J774 cells were incubated for up to 48 hr with 0·4–10 µg/ml EMA-CT DNA with or without Fugene 6. DNA uptake was assessed by flow cytometric analysis of the cells collected at different time-points. The levels of DNA uptake are represented as mean fluorescence intensity (MFI) for each group. There was a dose- and time-dependent increase of DNA uptake in the presence of Fugene 6.

    Article Snippet: Previous studies have shown that EC DNA is immunostimulatory, in contrast to CT DNA, which is inactive alone., In these experiments, J774 cells were treated with 10 µg/ml of ds and ss CT DNA or EC DNA alone or in complexes with Fugene 6 for up to 48 hr; supernatants were then examined for IL-12 and NO production.

    Techniques: Incubation, Flow Cytometry, Fluorescence

    Confocal microscopy analysis of intracellular distribution of CT DNA. J774 cells were simultaneously incubated with 1 µg/ml of EMA-labelled CT DNA in complexes with Fugene 6 (CT DNA) and 0·5 mg/ml Oregon green-labelled dextran (Dextran) at 37° for 12 hr. Internalized CT DNA (red-coloured) and dextran (green-coloured) were examined by confocal microscopy. CT DNA can be seen mostly co-localized (yellow-coloured) with dextran (Merged), indicating a fluid phase endocytosis pathway for DNA uptake (bar = 10 µm).

    Journal: Immunology

    Article Title: Effect of cytofectins on the immune response of murine macrophages to mammalian DNA

    doi: 10.1046/j.1365-2567.2003.01653.x

    Figure Lengend Snippet: Confocal microscopy analysis of intracellular distribution of CT DNA. J774 cells were simultaneously incubated with 1 µg/ml of EMA-labelled CT DNA in complexes with Fugene 6 (CT DNA) and 0·5 mg/ml Oregon green-labelled dextran (Dextran) at 37° for 12 hr. Internalized CT DNA (red-coloured) and dextran (green-coloured) were examined by confocal microscopy. CT DNA can be seen mostly co-localized (yellow-coloured) with dextran (Merged), indicating a fluid phase endocytosis pathway for DNA uptake (bar = 10 µm).

    Article Snippet: Previous studies have shown that EC DNA is immunostimulatory, in contrast to CT DNA, which is inactive alone., In these experiments, J774 cells were treated with 10 µg/ml of ds and ss CT DNA or EC DNA alone or in complexes with Fugene 6 for up to 48 hr; supernatants were then examined for IL-12 and NO production.

    Techniques: Confocal Microscopy, Incubation

    Effects of chloroquine on induction of NO by CT DNA–cytofectin complexes. J774 cells were pretreated with 1–4 µg/ml of chloroquine for 1 hr, followed by incubation with 10 µg/ml CT DNA in the presence of Fugene 6. EC DNA at 10 µg/ml and LPS at 1 µg/ml were used as controls. NO induction by CT DNA–cytofectin complexes was partially blocked by chloroquine. Whereas chloroquine blocked EC DNA-induced NO, it had a minimal effect on LPS activity. Error bars depict the standard deviations. The data are representative of at least three separate experiments. * P

    Journal: Immunology

    Article Title: Effect of cytofectins on the immune response of murine macrophages to mammalian DNA

    doi: 10.1046/j.1365-2567.2003.01653.x

    Figure Lengend Snippet: Effects of chloroquine on induction of NO by CT DNA–cytofectin complexes. J774 cells were pretreated with 1–4 µg/ml of chloroquine for 1 hr, followed by incubation with 10 µg/ml CT DNA in the presence of Fugene 6. EC DNA at 10 µg/ml and LPS at 1 µg/ml were used as controls. NO induction by CT DNA–cytofectin complexes was partially blocked by chloroquine. Whereas chloroquine blocked EC DNA-induced NO, it had a minimal effect on LPS activity. Error bars depict the standard deviations. The data are representative of at least three separate experiments. * P

    Article Snippet: Previous studies have shown that EC DNA is immunostimulatory, in contrast to CT DNA, which is inactive alone., In these experiments, J774 cells were treated with 10 µg/ml of ds and ss CT DNA or EC DNA alone or in complexes with Fugene 6 for up to 48 hr; supernatants were then examined for IL-12 and NO production.

    Techniques: Incubation, Activity Assay