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  • 99
    Thermo Fisher lipofectamine plus
    NGB reduces cyclin D1 expression and inhibits cell growth via merlin. (A) Protein and mRNA levels of cyclin D1 are inhibited by NGB. JS1 cells were transfected with increasing amounts of Flag-NGB and immunoblotted with indicated antibodies (panels 1 to 3). Total RNA was isolated from the cells and subjected to semiquantitative reverse transcription-PCR using primers specific for cyclin D1 and β-actin (panels 4 and 5). (B) Knockdown of NGB increases cyclin D1 level. HeLa cells were transfected with NGB-RNAi using <t>Lipofectamine.</t> Following 72 h of incubation, cells were lysed and immunoblotted with anti-NGB (top), -cyclin D1 (middle), and -actin (bottom) antibodies. (C) NGB-reduced cyclin D1 is abrogated by knockdown of merlin. JS1/NGB-59 cells were infected with lentivirus pLKO.1-shRNA/Nf2 and pLKO.1-puro vector and immunoblotted with indicated antibodies. (D and E) Merlin mediates NGB-inhibited cell growth and tumorigenicity. Indicated cells were seeded in 48-well plates at 0.2 × 10 5 /well. The cell number was counted daily for 3 days. The experiment was performed in triplicate (D). The cells were subcutaneously injected into nude mice (3 × 10 6 cells/mouse). Tumor volume was measured every 2 days. Data shown are representative of results from two independent experiments carried out with 16 mice each (8 mice/cell line) (E).
    Lipofectamine Plus, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 15798 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    lipofectamine plus - by Bioz Stars, 2020-09
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    99
    Thermo Fisher lipofectamine ltx plus reagent
    CD8 + T cell response to 13 T . lestoquardi antigens measured by IFNγ ELISA. Sheep 4247 fibroblasts were transfected with expression constructs of candidate antigen genes using either <t>Lipofectamine</t> <t>LTX</t> or Fugene transfection reagents for 48 h before the addition of effector cells for 72 h and IFNγ ELISA. The antigens were tested separately and data are presented as mean ± SD of biological repeats (n = 3).
    Lipofectamine Ltx Plus Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1551 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipofectamine ltx plus reagent/product/Thermo Fisher
    Average 99 stars, based on 1551 article reviews
    Price from $9.99 to $1999.99
    lipofectamine ltx plus reagent - by Bioz Stars, 2020-09
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    99
    Thermo Fisher lipofectamine plus reagents
    Uptake of cystine and CDDP resistance in A2780 cells transfected with cDNAs for xCT and 4F2hc. A2780 cells were cultured in the dish for 24 h and then they were transfected with pEGFP-C1 (Control) or cotransfected with pEGFP-C1/xCT and pEGFP-C1/4F2hc (x c − ) using <t>Lipofectamine</t> Plus™. After transfection, the cells were incubated for 33 h and the rate of cystine (0.05 m M ) uptake was measured ( A ) or CDDP resistance was examined ( B ). CDDP resistance was estimated by measuring viability of the cells treated with 50 μ M CDDP in fresh medium for the time indicated. Values are means±s.d. ( n =6). * Significantly greater than the corresponding control, P
    Lipofectamine Plus Reagents, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1209 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipofectamine plus reagents/product/Thermo Fisher
    Average 99 stars, based on 1209 article reviews
    Price from $9.99 to $1999.99
    lipofectamine plus reagents - by Bioz Stars, 2020-09
    99/100 stars
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    99
    Thermo Fisher lipofectamine plus transfection reagent
    The effects of CagA on cell growth and HER-2 expression. (A) Cell growth as measured by the MTT assay in AGS, MKN1, and HFE-145 cells after <t>transfection.</t> CagA transfected cells showed significantly increased growth. (B C) Effect of CagA on HER-2 DNA copy number and mRNA expression in AGS, MKN1, and HFE-145 cells. Ectopic CagA expression induced an increase of HER-2 DNA copy number and mRNA transcript expression. (D) HER-2 protein expression measured by Western blot in CagA transfected AGS, MKN1, and HFE-145 cells. Increased expression of the HER-2 protein was observed in CagA transfected cells compared to those in mock (vector + <t>Lipofectamine)</t> transfected cells. (E F) The gastric mucosae of H. pylori infected C57BL/6 mice showed increased HER-2 DNA copy number and protein expression. (G) CagA or H 2 O 2 treatment significantly increased ROS production, but treatment with the antioxidant TEMPOL reverted the CagA- and H 2 O 2 -induced ROS level in AGS,MKN1, and HFE-145 cells. (H) CagA or H 2 O 2 treatment significantly increased HER-2 DNA copy number, whereas TEMPOL reverted the H 2 O 2 -induced HER-2 copy number change in AGS cells.
    Lipofectamine Plus Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1874 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lipofectamine plus transfection reagent/product/Thermo Fisher
    Average 99 stars, based on 1874 article reviews
    Price from $9.99 to $1999.99
    lipofectamine plus transfection reagent - by Bioz Stars, 2020-09
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    Image Search Results


    NGB reduces cyclin D1 expression and inhibits cell growth via merlin. (A) Protein and mRNA levels of cyclin D1 are inhibited by NGB. JS1 cells were transfected with increasing amounts of Flag-NGB and immunoblotted with indicated antibodies (panels 1 to 3). Total RNA was isolated from the cells and subjected to semiquantitative reverse transcription-PCR using primers specific for cyclin D1 and β-actin (panels 4 and 5). (B) Knockdown of NGB increases cyclin D1 level. HeLa cells were transfected with NGB-RNAi using Lipofectamine. Following 72 h of incubation, cells were lysed and immunoblotted with anti-NGB (top), -cyclin D1 (middle), and -actin (bottom) antibodies. (C) NGB-reduced cyclin D1 is abrogated by knockdown of merlin. JS1/NGB-59 cells were infected with lentivirus pLKO.1-shRNA/Nf2 and pLKO.1-puro vector and immunoblotted with indicated antibodies. (D and E) Merlin mediates NGB-inhibited cell growth and tumorigenicity. Indicated cells were seeded in 48-well plates at 0.2 × 10 5 /well. The cell number was counted daily for 3 days. The experiment was performed in triplicate (D). The cells were subcutaneously injected into nude mice (3 × 10 6 cells/mouse). Tumor volume was measured every 2 days. Data shown are representative of results from two independent experiments carried out with 16 mice each (8 mice/cell line) (E).

    Journal: Molecular and Cellular Biology

    Article Title: Identification and Characterization of Putative Tumor Suppressor NGB, a GTP-Binding Protein That Interacts with the Neurofibromatosis 2 Protein ▿

    doi: 10.1128/MCB.00572-06

    Figure Lengend Snippet: NGB reduces cyclin D1 expression and inhibits cell growth via merlin. (A) Protein and mRNA levels of cyclin D1 are inhibited by NGB. JS1 cells were transfected with increasing amounts of Flag-NGB and immunoblotted with indicated antibodies (panels 1 to 3). Total RNA was isolated from the cells and subjected to semiquantitative reverse transcription-PCR using primers specific for cyclin D1 and β-actin (panels 4 and 5). (B) Knockdown of NGB increases cyclin D1 level. HeLa cells were transfected with NGB-RNAi using Lipofectamine. Following 72 h of incubation, cells were lysed and immunoblotted with anti-NGB (top), -cyclin D1 (middle), and -actin (bottom) antibodies. (C) NGB-reduced cyclin D1 is abrogated by knockdown of merlin. JS1/NGB-59 cells were infected with lentivirus pLKO.1-shRNA/Nf2 and pLKO.1-puro vector and immunoblotted with indicated antibodies. (D and E) Merlin mediates NGB-inhibited cell growth and tumorigenicity. Indicated cells were seeded in 48-well plates at 0.2 × 10 5 /well. The cell number was counted daily for 3 days. The experiment was performed in triplicate (D). The cells were subcutaneously injected into nude mice (3 × 10 6 cells/mouse). Tumor volume was measured every 2 days. Data shown are representative of results from two independent experiments carried out with 16 mice each (8 mice/cell line) (E).

    Article Snippet: Cell culture media, protein A/G beads, and Lipofectamine Plus were from Invitrogen (Carlsbad, CA).

    Techniques: Expressing, Transfection, Isolation, Polymerase Chain Reaction, Incubation, Infection, shRNA, Plasmid Preparation, Injection, Mouse Assay

    SOD2 knockdown inhibits both the migration and invasion abilities of TSCC To characterize the role of SOD2 in aiding metastasis, the plasmid containing SOD2 shRNA was transiently transfected into UM1 cells using Lipofectamine Plus reagent. Cells were tested 24 h post-transfection. (A) Significant reduction of SOD2 protein levels and activities were observed in the SOD2 shRNA-transfected UM1 cells compared to the vector control transfected cells. *: P

    Journal: Free radical biology & medicine

    Article Title: Manganese superoxide dismutase induces migration and invasion of tongue squamous cell carcinoma via H2O2-dependent Snail signaling

    doi: 10.1016/j.freeradbiomed.2012.04.031

    Figure Lengend Snippet: SOD2 knockdown inhibits both the migration and invasion abilities of TSCC To characterize the role of SOD2 in aiding metastasis, the plasmid containing SOD2 shRNA was transiently transfected into UM1 cells using Lipofectamine Plus reagent. Cells were tested 24 h post-transfection. (A) Significant reduction of SOD2 protein levels and activities were observed in the SOD2 shRNA-transfected UM1 cells compared to the vector control transfected cells. *: P

    Article Snippet: The constructed plasmids were transiently transfected into UM1 cells using Lipofectamine Plus reagent (Invitrogen, CA, USA), according to the manufacturer's instructions[ ].

    Techniques: Migration, Plasmid Preparation, shRNA, Transfection

    Snail signaling contributes to SOD2-induced migration and invasion of TSCC To characterize the role of Snail signaling in SOD2-induced metastasis of TSCC, western blot analysis was used with actin as the loading control. Plasmids containing SOD2 shRNA were transiently transfected into UM1 cells using Lipofectamine Plus reagent. Cells were tested 24 h post-transfection. The UM2 cells or SOD2 shRNA-transfected UM1 cells were treated with 100μM H 2 O 2 for 24 h. (A) UM1 cells displayed an increase in Snai1, Snai2, MMP-1, ERK1/2 and pERK1/2 protein levels, and decreased protein levels of E-cadtherin compared to UM2 cells. (B) UM1 cells displayed decreased snai1, snai2, MMP-1, ERK1/2 and pERK1/2 protein levels and increased E-cadtherin protein levels upon SOD2 knockdown. (C, D) The addition of H 2 O 2 increased the protein levels of Snai1, Snai2, MMP-1, ERK1/2 and pERK1/2 and decreased protein levels of E-cadtherin in both UM2 cells and the SOD2 shRNA-transfected UMl cells.

    Journal: Free radical biology & medicine

    Article Title: Manganese superoxide dismutase induces migration and invasion of tongue squamous cell carcinoma via H2O2-dependent Snail signaling

    doi: 10.1016/j.freeradbiomed.2012.04.031

    Figure Lengend Snippet: Snail signaling contributes to SOD2-induced migration and invasion of TSCC To characterize the role of Snail signaling in SOD2-induced metastasis of TSCC, western blot analysis was used with actin as the loading control. Plasmids containing SOD2 shRNA were transiently transfected into UM1 cells using Lipofectamine Plus reagent. Cells were tested 24 h post-transfection. The UM2 cells or SOD2 shRNA-transfected UM1 cells were treated with 100μM H 2 O 2 for 24 h. (A) UM1 cells displayed an increase in Snai1, Snai2, MMP-1, ERK1/2 and pERK1/2 protein levels, and decreased protein levels of E-cadtherin compared to UM2 cells. (B) UM1 cells displayed decreased snai1, snai2, MMP-1, ERK1/2 and pERK1/2 protein levels and increased E-cadtherin protein levels upon SOD2 knockdown. (C, D) The addition of H 2 O 2 increased the protein levels of Snai1, Snai2, MMP-1, ERK1/2 and pERK1/2 and decreased protein levels of E-cadtherin in both UM2 cells and the SOD2 shRNA-transfected UMl cells.

    Article Snippet: The constructed plasmids were transiently transfected into UM1 cells using Lipofectamine Plus reagent (Invitrogen, CA, USA), according to the manufacturer's instructions[ ].

    Techniques: Migration, Western Blot, shRNA, Transfection

    Dominant-negative mutants of p65 (Δp65) and C/EBP β (ΔC/EBP β ) inhibit MBP-primed T cell-induced expression of proinflammatory cytokines in BV-2 microglial cells Cells were transfected with 0.5 μ g of either Δp65 or ΔC/EBP β using LipofectAMINE-plus. After 24 h of transfection, cells were stimulated with MBP-primed T cells (0.5:1 of T cell:glia) under serum-free condition. A , after 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free medium for another 23 h. Supernatants were used to assay IL-1 β and TNF- α . Data are mean ± S.D. of three different experiments. B , after 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free media for another 3 h. The expression of different cytokines was analyzed by a gene array kit (GEArray). The relative expression of different cytokines (cytokines/glyceraldehyde-3-phosphate dehydrogenase ( GAPDH )) was measured after scanning the bands with a Fluor Chem 8800 Imaging System. Data are mean ± S.D. of three different experiments.

    Journal: The Journal of biological chemistry

    Article Title: Role of Very-late Antigen-4 (VLA-4) in Myelin Basic Protein-primed T Cell Contact-induced Expression of Proinflammatory Cytokines in Microglial Cells *

    doi: 10.1074/jbc.M301789200

    Figure Lengend Snippet: Dominant-negative mutants of p65 (Δp65) and C/EBP β (ΔC/EBP β ) inhibit MBP-primed T cell-induced expression of proinflammatory cytokines in BV-2 microglial cells Cells were transfected with 0.5 μ g of either Δp65 or ΔC/EBP β using LipofectAMINE-plus. After 24 h of transfection, cells were stimulated with MBP-primed T cells (0.5:1 of T cell:glia) under serum-free condition. A , after 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free medium for another 23 h. Supernatants were used to assay IL-1 β and TNF- α . Data are mean ± S.D. of three different experiments. B , after 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free media for another 3 h. The expression of different cytokines was analyzed by a gene array kit (GEArray). The relative expression of different cytokines (cytokines/glyceraldehyde-3-phosphate dehydrogenase ( GAPDH )) was measured after scanning the bands with a Fluor Chem 8800 Imaging System. Data are mean ± S.D. of three different experiments.

    Article Snippet: To assay the transcriptional activity of NF- κ B and C/EBP β , cells at 50–60% confluence in 12-well plates were transfected with 0.5 μ g of either PBIIX-Luc, an NF- κ B-dependent reporter construct , or pC/EBP β -Luc, an C/EBP β -dependent reporter construct , using the LipofectAMINE Plus method (Invitrogen) ( - ).

    Techniques: Dominant Negative Mutation, Expressing, Transfection, Incubation, Imaging

    MBP-primed T cells induce the activation of NF- κ B in BV-2 microglial cells A , cells incubated in serum-free DMEM/F-12 were stimulated with MBP-primed T cells (0.5:1 of T cell:glia). After different minutes of stimulation, culture dishes were shaken and washed three times with HBSS. Adherent cells were taken out to prepare nuclear extracts, and nuclear proteins were used for EMSA as described under “Materials and Methods.” The upper , middle , and lower arrows indicate the induced NF- κ B band, nonspecific band, and the unbound probe, respectively. B , cells plated at 50–60% confluence in 12-well plates were cotransfected with 0.5 μ g of PBIIX-Luc and 25 ng of pRL-TK using LipofectAMINE Plus as described under “Materials and Methods.” After 24 h of transfection, BV-2 cells were stimulated with different concentrations of MBP-primed T cells. After 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free medium for another 5 h. Firefly and Renilla luciferase activities were obtained by analyzing total cell extract of BV-2 cells as described under “Materials and Methods.” Data are mean ± S.D. of three different experiments.

    Journal: The Journal of biological chemistry

    Article Title: Role of Very-late Antigen-4 (VLA-4) in Myelin Basic Protein-primed T Cell Contact-induced Expression of Proinflammatory Cytokines in Microglial Cells *

    doi: 10.1074/jbc.M301789200

    Figure Lengend Snippet: MBP-primed T cells induce the activation of NF- κ B in BV-2 microglial cells A , cells incubated in serum-free DMEM/F-12 were stimulated with MBP-primed T cells (0.5:1 of T cell:glia). After different minutes of stimulation, culture dishes were shaken and washed three times with HBSS. Adherent cells were taken out to prepare nuclear extracts, and nuclear proteins were used for EMSA as described under “Materials and Methods.” The upper , middle , and lower arrows indicate the induced NF- κ B band, nonspecific band, and the unbound probe, respectively. B , cells plated at 50–60% confluence in 12-well plates were cotransfected with 0.5 μ g of PBIIX-Luc and 25 ng of pRL-TK using LipofectAMINE Plus as described under “Materials and Methods.” After 24 h of transfection, BV-2 cells were stimulated with different concentrations of MBP-primed T cells. After 1 h of stimulation, T cells were removed followed by the incubation of adherent BV-2 cells in serum-free medium for another 5 h. Firefly and Renilla luciferase activities were obtained by analyzing total cell extract of BV-2 cells as described under “Materials and Methods.” Data are mean ± S.D. of three different experiments.

    Article Snippet: To assay the transcriptional activity of NF- κ B and C/EBP β , cells at 50–60% confluence in 12-well plates were transfected with 0.5 μ g of either PBIIX-Luc, an NF- κ B-dependent reporter construct , or pC/EBP β -Luc, an C/EBP β -dependent reporter construct , using the LipofectAMINE Plus method (Invitrogen) ( - ).

    Techniques: Activation Assay, Incubation, Transfection, Luciferase

    FIP200 interacts with PIASy in vivo. (A) HEK293T cells were transfected with a combination of empty Flag/HA-vector (Ctrl), wild-type (wt) Flag/HA-PIASy, C342F mutant (mut) Flag/HA-PIASy, and Flag-FIP200 as indicated. At 24 h after transfection, 100 μg

    Journal: Molecular and Cellular Biology

    Article Title: Spatial Interplay between PIASy and FIP200 in the Regulation of Signal Transduction and Transcriptional Activity

    doi: 10.1128/MCB.01210-07

    Figure Lengend Snippet: FIP200 interacts with PIASy in vivo. (A) HEK293T cells were transfected with a combination of empty Flag/HA-vector (Ctrl), wild-type (wt) Flag/HA-PIASy, C342F mutant (mut) Flag/HA-PIASy, and Flag-FIP200 as indicated. At 24 h after transfection, 100 μg

    Article Snippet: Cells were transfected with 0.3 μg p21-luciferase and 0.01 μg pCMV-β-gal reporters plus expression constructs by using Lipofectamine and Plus transfection reagents (Invitrogen) according to the manufacturers’ instructions. β-Galactosidase and luciferase activities were measured with the Galacto-Star (Tropix) and Luciferase Assay Systems (Promega) kits according to the manufacturers’ instructions.

    Techniques: In Vivo, Transfection, Plasmid Preparation, Mutagenesis

    CD8 + T cell response to 13 T . lestoquardi antigens measured by IFNγ ELISA. Sheep 4247 fibroblasts were transfected with expression constructs of candidate antigen genes using either Lipofectamine LTX or Fugene transfection reagents for 48 h before the addition of effector cells for 72 h and IFNγ ELISA. The antigens were tested separately and data are presented as mean ± SD of biological repeats (n = 3).

    Journal: PLoS ONE

    Article Title: Identification of Theileria lestoquardi Antigens Recognized by CD8+ T Cells

    doi: 10.1371/journal.pone.0162571

    Figure Lengend Snippet: CD8 + T cell response to 13 T . lestoquardi antigens measured by IFNγ ELISA. Sheep 4247 fibroblasts were transfected with expression constructs of candidate antigen genes using either Lipofectamine LTX or Fugene transfection reagents for 48 h before the addition of effector cells for 72 h and IFNγ ELISA. The antigens were tested separately and data are presented as mean ± SD of biological repeats (n = 3).

    Article Snippet: Transfection was carried out either with Lipofectamine LTX & Plus Reagent (Invitrogen) or Fugene (Promega) at reagent (μl): DNA (μg) ratios of 5:1 and 3:1, respectively.

    Techniques: Enzyme-linked Immunosorbent Assay, Transfection, Expressing, Construct

    Characterization of commercial vectors (i.e., jetPRIME, GeneIn and Lipofectamine LTX/PLUS) in terms of size and charge. A) Size analysis of nanoparticles formed through complexation of each vector with 0.5 µg of pEGFP. The nanoparticles were formed following manufacturers’ protocols. B) Surface charge (Zeta potential) analysis of the same nanoparticles.

    Journal: Acta biomaterialia

    Article Title: Evaluation of Genotoxicity and Mutagenic Effects of Vector/DNA Nanocomplexes in Transfected Mesenchymal Stem Cells by Flow Cytometry

    doi: 10.1016/j.actbio.2018.05.029

    Figure Lengend Snippet: Characterization of commercial vectors (i.e., jetPRIME, GeneIn and Lipofectamine LTX/PLUS) in terms of size and charge. A) Size analysis of nanoparticles formed through complexation of each vector with 0.5 µg of pEGFP. The nanoparticles were formed following manufacturers’ protocols. B) Surface charge (Zeta potential) analysis of the same nanoparticles.

    Article Snippet: Gene transfer reagents, including GeneIn™ (AMSBIO, CA), Lipofectamine® LTX/PLUS™ (Thermo Fisher Scientific Inc.), and jetPRIME® (Polyplus-transfection® SA), were purchased from commercially available sources.

    Techniques: Plasmid Preparation

    Transfection of the GFP plasmid in HEK-293S cells with different reagents and the expression of GFP at 33°C. Bright-field and fluorescence images (34.8x magnification, scale bar = 50 μm) of HEK-293S cells taken at the 48th hour after transfection. Transfection of HEK-293S cells as in images (A) to (D) was carried out using Lipofectamine 2000, Lipofectamine LTX PLUS, Metafectene EASY with Opti-MEM and Metafectene EASY with DMEM with 10% FBS, respectively. In each of the transfections, 2 μg GFP plasmid for a 35 mm Petri dish was used. On day 1, all transfected dishes were incubated at 37°C; on day 2, these dishes were transferred to a 33°C incubator. Based on these images, the transfection efficiency for transient expression of GFP was determined to be ~64%, ~61%, ~32% and ~57% from (A) to (D), respectively. (E) The green cells were categorized into high (intensity > 200), middle (intensity 100–200), and low (intensity 29–100) three groups. The black color indicates cells visible in the bright view but can’t been observed under UV (intensity

    Journal: PLoS ONE

    Article Title: Enhancing Protein Expression in HEK-293 Cells by Lowering Culture Temperature

    doi: 10.1371/journal.pone.0123562

    Figure Lengend Snippet: Transfection of the GFP plasmid in HEK-293S cells with different reagents and the expression of GFP at 33°C. Bright-field and fluorescence images (34.8x magnification, scale bar = 50 μm) of HEK-293S cells taken at the 48th hour after transfection. Transfection of HEK-293S cells as in images (A) to (D) was carried out using Lipofectamine 2000, Lipofectamine LTX PLUS, Metafectene EASY with Opti-MEM and Metafectene EASY with DMEM with 10% FBS, respectively. In each of the transfections, 2 μg GFP plasmid for a 35 mm Petri dish was used. On day 1, all transfected dishes were incubated at 37°C; on day 2, these dishes were transferred to a 33°C incubator. Based on these images, the transfection efficiency for transient expression of GFP was determined to be ~64%, ~61%, ~32% and ~57% from (A) to (D), respectively. (E) The green cells were categorized into high (intensity > 200), middle (intensity 100–200), and low (intensity 29–100) three groups. The black color indicates cells visible in the bright view but can’t been observed under UV (intensity

    Article Snippet: A number of transfection reagents were used: calcium phosphate [ ], Lipofectamine 2000 (Invitrogen, Cat. No. 18324–111, Carlsbad, CA), Lipofectamine LTX & PLUS (Invitrogen, Cat. No. 15338030) and Metafectene EASY (Biontex Laboratories GmbH, Cat. No. T090-10, Munich, Germany).

    Techniques: Transfection, Plasmid Preparation, Expressing, Fluorescence, Incubation

    Uptake of cystine and CDDP resistance in A2780 cells transfected with cDNAs for xCT and 4F2hc. A2780 cells were cultured in the dish for 24 h and then they were transfected with pEGFP-C1 (Control) or cotransfected with pEGFP-C1/xCT and pEGFP-C1/4F2hc (x c − ) using Lipofectamine Plus™. After transfection, the cells were incubated for 33 h and the rate of cystine (0.05 m M ) uptake was measured ( A ) or CDDP resistance was examined ( B ). CDDP resistance was estimated by measuring viability of the cells treated with 50 μ M CDDP in fresh medium for the time indicated. Values are means±s.d. ( n =6). * Significantly greater than the corresponding control, P

    Journal: British Journal of Cancer

    Article Title: Role of cystine transport in intracellular glutathione level and cisplatin resistance in human ovarian cancer cell lines

    doi: 10.1038/sj.bjc.6600786

    Figure Lengend Snippet: Uptake of cystine and CDDP resistance in A2780 cells transfected with cDNAs for xCT and 4F2hc. A2780 cells were cultured in the dish for 24 h and then they were transfected with pEGFP-C1 (Control) or cotransfected with pEGFP-C1/xCT and pEGFP-C1/4F2hc (x c − ) using Lipofectamine Plus™. After transfection, the cells were incubated for 33 h and the rate of cystine (0.05 m M ) uptake was measured ( A ) or CDDP resistance was examined ( B ). CDDP resistance was estimated by measuring viability of the cells treated with 50 μ M CDDP in fresh medium for the time indicated. Values are means±s.d. ( n =6). * Significantly greater than the corresponding control, P

    Article Snippet: Transient transfections were performed using Lipofectamine Plus™ reagents (Invitrogen Corp., Carlsbad, CA, USA) according to the manufacturer's instruction.

    Techniques: Transfection, Cell Culture, Incubation

    The effects of CagA on cell growth and HER-2 expression. (A) Cell growth as measured by the MTT assay in AGS, MKN1, and HFE-145 cells after transfection. CagA transfected cells showed significantly increased growth. (B C) Effect of CagA on HER-2 DNA copy number and mRNA expression in AGS, MKN1, and HFE-145 cells. Ectopic CagA expression induced an increase of HER-2 DNA copy number and mRNA transcript expression. (D) HER-2 protein expression measured by Western blot in CagA transfected AGS, MKN1, and HFE-145 cells. Increased expression of the HER-2 protein was observed in CagA transfected cells compared to those in mock (vector + Lipofectamine) transfected cells. (E F) The gastric mucosae of H. pylori infected C57BL/6 mice showed increased HER-2 DNA copy number and protein expression. (G) CagA or H 2 O 2 treatment significantly increased ROS production, but treatment with the antioxidant TEMPOL reverted the CagA- and H 2 O 2 -induced ROS level in AGS,MKN1, and HFE-145 cells. (H) CagA or H 2 O 2 treatment significantly increased HER-2 DNA copy number, whereas TEMPOL reverted the H 2 O 2 -induced HER-2 copy number change in AGS cells.

    Journal: Gene

    Article Title: The effect of Helicobacter pylori CagA on the HER-2 copy number and expression in gastric cancer

    doi: 10.1016/j.gene.2014.05.064

    Figure Lengend Snippet: The effects of CagA on cell growth and HER-2 expression. (A) Cell growth as measured by the MTT assay in AGS, MKN1, and HFE-145 cells after transfection. CagA transfected cells showed significantly increased growth. (B C) Effect of CagA on HER-2 DNA copy number and mRNA expression in AGS, MKN1, and HFE-145 cells. Ectopic CagA expression induced an increase of HER-2 DNA copy number and mRNA transcript expression. (D) HER-2 protein expression measured by Western blot in CagA transfected AGS, MKN1, and HFE-145 cells. Increased expression of the HER-2 protein was observed in CagA transfected cells compared to those in mock (vector + Lipofectamine) transfected cells. (E F) The gastric mucosae of H. pylori infected C57BL/6 mice showed increased HER-2 DNA copy number and protein expression. (G) CagA or H 2 O 2 treatment significantly increased ROS production, but treatment with the antioxidant TEMPOL reverted the CagA- and H 2 O 2 -induced ROS level in AGS,MKN1, and HFE-145 cells. (H) CagA or H 2 O 2 treatment significantly increased HER-2 DNA copy number, whereas TEMPOL reverted the H 2 O 2 -induced HER-2 copy number change in AGS cells.

    Article Snippet: AGS,MKN1 and HFE-145 cells were transfected in 60 mm-diameter dishes with expression plasmids (2 µg total DNA) using a Lipofectamine Plus transfection reagent (Invitrogen) according to the manufacturer's recommendations.

    Techniques: Expressing, MTT Assay, Transfection, Western Blot, Plasmid Preparation, Infection, Mouse Assay