lipid transfection  (Qiagen)


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    Structured Review

    Qiagen lipid transfection
    Lipid Transfection, supplied by Qiagen, 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/result/lipid transfection/product/Qiagen
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    lipid transfection - by Bioz Stars, 2021-06
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    Related Articles

    Negative Control:

    Article Title: MicroRNA-31-3p Is Involved in Substance P (SP)-Associated Inflammation in Human Colonic Epithelial Cells and Experimental Colitis
    Article Snippet: Inhibitors of miR-31-3p (catalog number 4464084), negative anti-miRNA controls (catalog number 4464076), a miR-31-3p mimic (catalog number 4464066), and mimic miRNA controls (catalog number 4464058) were purchased from Life Technologies. .. Mouse anti–miR-31-3p and its negative control (sequence of anti–miR-31-3p is 5′-AATATGTTGGCATAGC-3′, sequence of anti-miRNA control is 5′-ACGTCTATACGCCCA-3′) were purchased from Qiagen (Germantown, MD). .. Lipid-based siPORTNeoFX Transfection Agent (AM4511) was purchased from Ambion (Canoga Park, CA).

    Sequencing:

    Article Title: MicroRNA-31-3p Is Involved in Substance P (SP)-Associated Inflammation in Human Colonic Epithelial Cells and Experimental Colitis
    Article Snippet: Inhibitors of miR-31-3p (catalog number 4464084), negative anti-miRNA controls (catalog number 4464076), a miR-31-3p mimic (catalog number 4464066), and mimic miRNA controls (catalog number 4464058) were purchased from Life Technologies. .. Mouse anti–miR-31-3p and its negative control (sequence of anti–miR-31-3p is 5′-AATATGTTGGCATAGC-3′, sequence of anti-miRNA control is 5′-ACGTCTATACGCCCA-3′) were purchased from Qiagen (Germantown, MD). .. Lipid-based siPORTNeoFX Transfection Agent (AM4511) was purchased from Ambion (Canoga Park, CA).

    Transfection:

    Article Title: Nebulin Interacts with CapZ and Regulates Thin Filament Architecture within the Z-Disc
    Article Snippet: As a control, a random siRNA was also generated (target cDNA sequence: 5′-CTC GAC TAG AGT CTG TCT A-3′). .. Skeletal myotubes were transfected with 50 nM siRNA using the lipid-based reagent Effectene (QIAGEN, Valencia, CA) according to the manufacturer's instructions 12 to 24 h after plating. .. Two to 3 d after transfection, the cells were incubated in relaxing buffer [150 mM KCl, 5 mM MgCl2 , 10 mM 3-( N -morpholino)propanesulfonic acid, pH 7.4, 1 mM EGTA, and 4 mM ATP] for 15 min, and then they were fixed with 0.5–2% paraformaldehyde in relaxing buffer for 15 min.

    Article Title: High-Throughput Transcriptomic and RNAi Analysis Identifies AIM1, ERGIC1, TMED3 and TPX2 as Potential Drug Targets in Prostate Cancer
    Article Snippet: .. For the RNAi studies, four siRNAs per gene (HP GenomeWide, Qiagen) were plated onto 384-well plates (Greiner Bio-One, Frickenhausen, Germany), followed by addition of the transfection agent (siLentFect lipid reagent; Bio-Rad Laboratories, Hercules, CA) in Opti-MEM medium (Invitrogen) and an appropriate quantity of cells (1500–2000 per well), using automated liquid handling robot (Hamilton) and liquid dispenser (ThermoFisher). .. The final siRNA concentration was 13 nM.

    Article Title: The Carboxy-Terminal Tail of Human Cytomegalovirus (HCMV) US28 Regulates both Chemokine-Independent and Chemokine-Dependent Signaling in HCMV-Infected Cells
    Article Snippet: The sequences of the US28/1-314 recombinants including the US28 locus and regions surrounding the recombination site were confirmed by automated ABI DNA sequencing. .. For reconstitution of recombinant viruses, 2 × 105 MRC-5 cells were plated in six-well plates and transfected with 2 μg of FIX-BAC DNAs by using either Superfect (Qiagen) or Transit IT (Mirus) lipid transfection reagents according to the manufacturer's protocol. ..

    Small Interfering RNA:

    Article Title: Formyl Peptide Receptor 1 Modulates Endothelial Cell Functions by NADPH Oxidase-Dependent VEGFR2 Transactivation
    Article Snippet: .. Short interfering RNA experiments were performed incubating 4 × 105 cells with 5 nM siRNAs for 12 hours, in DMEM containing 10% FBS and 20 μ l of HiPerFect (Qiagen, Hiden, Germany). ..

    Binding Assay:

    Article Title: Inhibition of T-Cell Receptor Signal Transduction and Viral Expression by the Linker for Activation of T Cells-Interacting p12I Protein of Human T-Cell Leukemia/Lymphoma Virus Type 1 ▿
    Article Snippet: .. Binding of p12I to LAT-Myc was also confirmed in 293T cells, where antibody to p12I coprecipitated LAT-Myc, anti-Myc antibody coprecipitated p12I , and again, the anti-AU1 antibody coprecipitated neither (data not shown). ..

    Recombinant:

    Article Title: The Carboxy-Terminal Tail of Human Cytomegalovirus (HCMV) US28 Regulates both Chemokine-Independent and Chemokine-Dependent Signaling in HCMV-Infected Cells
    Article Snippet: The sequences of the US28/1-314 recombinants including the US28 locus and regions surrounding the recombination site were confirmed by automated ABI DNA sequencing. .. For reconstitution of recombinant viruses, 2 × 105 MRC-5 cells were plated in six-well plates and transfected with 2 μg of FIX-BAC DNAs by using either Superfect (Qiagen) or Transit IT (Mirus) lipid transfection reagents according to the manufacturer's protocol. ..

    other:

    Article Title: SREBP-1c/MicroRNA 33b Genomic Loci Control Adipocyte Differentiation
    Article Snippet: These findings suggest that the induction of miR-33b during the later stages of adipogenesis may serve as a mechanism to prevent excessive adipocyte hypertrophy due to overaccumulation of lipids.

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    Qiagen lipid mediated transient transfections
    DNA sequence analysis of clones created by HEG cleavage and non-homologous repair. ( a ) Clones of pBC/SacRB P1 isolated from Sua4.0 cells after <t>co-transfection</t> with I-PpoI expression vector pEGFP-Ppo. ( b ) Clones of pBC/SacRB S1 isolated from Sua4.0 cells (top) and G3 embryos (bottom) after co-transfection/injection with I-SceI expression vector pP[v+,70I-SceI]. The native 15 bp minimal I-PpoI and 18-bp I-SceI recognition sites are shown on top of the isolated clones in the context of the SacRB gene. Deleted nucleotides are indicated by dashes (-), inserted nucleotides are underlined. If deletions extend beyond the EcoRI sites flanking the HEG recognition sites the number of base pairs deleted is indicated and the first 3 bp after the deletion are shown in brackets. The shaded area indicates the 4 bp between the HEG cleavage positions on both DNA strands. Larger insertions are marked by vertical bars: (*) Insertion of 66 bp partially homologous to A. gambiae genome. (**) Insertion of 43 bp. B, BamHI; E, EcoRI; H, HindIII.
    Lipid Mediated Transient Transfections, supplied by Qiagen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen nextgeneration lipid reagent attractene transfection reagent
    Evaluation of <t>transfection</t> efficiency of B16-F10 cells transfected with use of carrier:pDNA complexes. (a) Percentage of GFP-positive cells transfected with complexes: AP:pGFP at different N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15) and AP-15/DOPE:pGFP, AP-15/DOPE/DMEM:pGFP lipoplexes containing 2.5 μg of lipids/μg of pGFP, analysed by FACS; Ap- significant difference from AP-15 treatment, At- significant difference from <t>Attractene</t> treatment, L- significant difference from Lipofectamine treatment, P- significant difference from PEI treatment, (b) Activity of β-galactosidase in cells transfected with complexes: AP-11:pLacZ, AP-15:pLacZ, PEI:pLacZ, at r = 2.0–2.5 N/P ratio, studied by β-Gal test; *P
    Nextgeneration Lipid Reagent Attractene Transfection Reagent, supplied by Qiagen, 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/result/nextgeneration lipid reagent attractene transfection reagent/product/Qiagen
    Average 86 stars, based on 1 article reviews
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    86
    Qiagen cationic lipid transfection reagent
    Mutational disruption of NK1R-mediated sequestration of β-arrestin prevents inhibition of F-MOR endocytosis in N2A cells (A) Confocal micrographs of N2A cells co-expressing either full-length HA-NK1R (left panels, red) or truncated HA-NK1R (right panels, red) with β-arrestin2-EGFP (green). Incubation with 10μM SP in cells expressing the truncated HA-NK1R (355x) resulted in less co-localization with receptors in endosomal clusters and higher cytoplasmic β-arrestin2-EGFP distribution (left column), compared to β-arrestin2-EGFP distribution in the wild type HA-NK1R under the same conditions (right column). Scale bar, 10μm. (B) In N2A cells co-expressing F-MOR (red) and HA-NK1 355x receptors (green), both morphine and DAMGO were able to drive internalization of F-MORs even in the presence of substance P. Scale bar, 10μm. (C) Quantification using ratiometric staining of F-MORs in N2A cells co-expressing HA-NK1 355x revealed that co-incubation with SP produced comparable levels of F-MOR internalization compared to treatment with either MS or DG alone. Bar graphs represent mean internalization determined from ~50 cell bodies selected at random in the N2A cultures and averaged over four independent <t>transfections.</t> (D) Biochemical assay of F-MOR internalization by surface biotinylation confirmed that F-MOR internalization induced by either MS or DG (lanes 3 and 5) was not significantly inhibited by co-application of SP (lanes 4 and 6; n= 3 independent experiments).
    Cationic Lipid Transfection Reagent, supplied by Qiagen, 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/result/cationic lipid transfection reagent/product/Qiagen
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    Qiagen superfect
    Comparative transfection efficiency of FuGENE HD (FuGENE), X-tremeGENE (X-treme), <t>SuperFect</t> (Super), Lipofectamine 2000 (L-2000), Lipofectamine RNAiMAX (L-iMAX), INT, and mTat/PEI/INT using siRNA targeting β - actin in HSC-3 cells. β - actin mRNA was measured by QRT-PCR and then % remaining β - actin mRNA expression was calculated based on control as 100% ( a ). Cell viability of each of the reagent groups in HaCaT cells were evaluated by MTT assay under the same siRNA and transfection reagent volume ( b ). * p
    Superfect, supplied by Qiagen, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    superfect - by Bioz Stars, 2021-06
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    DNA sequence analysis of clones created by HEG cleavage and non-homologous repair. ( a ) Clones of pBC/SacRB P1 isolated from Sua4.0 cells after co-transfection with I-PpoI expression vector pEGFP-Ppo. ( b ) Clones of pBC/SacRB S1 isolated from Sua4.0 cells (top) and G3 embryos (bottom) after co-transfection/injection with I-SceI expression vector pP[v+,70I-SceI]. The native 15 bp minimal I-PpoI and 18-bp I-SceI recognition sites are shown on top of the isolated clones in the context of the SacRB gene. Deleted nucleotides are indicated by dashes (-), inserted nucleotides are underlined. If deletions extend beyond the EcoRI sites flanking the HEG recognition sites the number of base pairs deleted is indicated and the first 3 bp after the deletion are shown in brackets. The shaded area indicates the 4 bp between the HEG cleavage positions on both DNA strands. Larger insertions are marked by vertical bars: (*) Insertion of 66 bp partially homologous to A. gambiae genome. (**) Insertion of 43 bp. B, BamHI; E, EcoRI; H, HindIII.

    Journal: Nucleic Acids Research

    Article Title: Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos

    doi: 10.1093/nar/gkm632

    Figure Lengend Snippet: DNA sequence analysis of clones created by HEG cleavage and non-homologous repair. ( a ) Clones of pBC/SacRB P1 isolated from Sua4.0 cells after co-transfection with I-PpoI expression vector pEGFP-Ppo. ( b ) Clones of pBC/SacRB S1 isolated from Sua4.0 cells (top) and G3 embryos (bottom) after co-transfection/injection with I-SceI expression vector pP[v+,70I-SceI]. The native 15 bp minimal I-PpoI and 18-bp I-SceI recognition sites are shown on top of the isolated clones in the context of the SacRB gene. Deleted nucleotides are indicated by dashes (-), inserted nucleotides are underlined. If deletions extend beyond the EcoRI sites flanking the HEG recognition sites the number of base pairs deleted is indicated and the first 3 bp after the deletion are shown in brackets. The shaded area indicates the 4 bp between the HEG cleavage positions on both DNA strands. Larger insertions are marked by vertical bars: (*) Insertion of 66 bp partially homologous to A. gambiae genome. (**) Insertion of 43 bp. B, BamHI; E, EcoRI; H, HindIII.

    Article Snippet: In vivo HEG activity assays were performed by lipid-mediated transient transfections (Effecten, Qiagen) of 1–3 × 105 cells with 2 μg/ml (culture volume) recipient plasmid and 4 μg/ml donor plasmid.

    Techniques: Sequencing, Clone Assay, Isolation, Cotransfection, Expressing, Plasmid Preparation, Injection

    Cell proliferation analysis and morphology of I-PpoI and I-SceI expressing cells. ( a ) I-PpoI expression arrests proliferation of Sua 4.0 cells. Cells were co-transfected with HEG expression vectors and a GFP control in combination with pIB/V5-His which confers resistance to blasticidin and counted after 5 days of growth in medium supplemented with blasticidin S. Red bars indicate cells that were heat-shocked to induce expression of I-SceI. The dotted line indicates the 150 000 cells seeded for the experiment. ( b ) Sua 4.0 cells were transfected with pEGFP-Ppo, pEGFP-Ppo H98A and control pSL-Act-GFP. Forty-eight hour post-transfection, cells were fixed and stained with DAPI as well as phalloidin. White arrows indicate nucleoli of cells. TM, transmission.

    Journal: Nucleic Acids Research

    Article Title: Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos

    doi: 10.1093/nar/gkm632

    Figure Lengend Snippet: Cell proliferation analysis and morphology of I-PpoI and I-SceI expressing cells. ( a ) I-PpoI expression arrests proliferation of Sua 4.0 cells. Cells were co-transfected with HEG expression vectors and a GFP control in combination with pIB/V5-His which confers resistance to blasticidin and counted after 5 days of growth in medium supplemented with blasticidin S. Red bars indicate cells that were heat-shocked to induce expression of I-SceI. The dotted line indicates the 150 000 cells seeded for the experiment. ( b ) Sua 4.0 cells were transfected with pEGFP-Ppo, pEGFP-Ppo H98A and control pSL-Act-GFP. Forty-eight hour post-transfection, cells were fixed and stained with DAPI as well as phalloidin. White arrows indicate nucleoli of cells. TM, transmission.

    Article Snippet: In vivo HEG activity assays were performed by lipid-mediated transient transfections (Effecten, Qiagen) of 1–3 × 105 cells with 2 μg/ml (culture volume) recipient plasmid and 4 μg/ml donor plasmid.

    Techniques: Expressing, Transfection, Activated Clotting Time Assay, Staining, Transmission Assay

    I-SceI and I-PpoI expression constructs and their activity in A. gambiae cells. ( a ) Maps of HEG expression and target vectors used in the interplasmid activity assay. CMV, cytomegalovirus promoter; Hsp70, Drosophila heatshock protein 70 promoter; eGFP enhanced green fluorescent protein; SacRB , levansucrase gene; X, XbaI; B, BamHI; E, EcoRI; H, HindIII; Xh, XhoI; N, NcoI; A, AscI; S, SalI; Nh, NheI; Kan R , kanamycin resistance cassette; Cam R , chloramphenicol resistance cassette; Tet R , tetracycline resistance cassette; Amp R , ampicilin resistance cassette; NLS, nuclear localization signal. ( b ) Analysis of I-SceI and I-PpoI activity in Sua4.0 cells by Southern blot. Cells were co-transfected with I-SceI or I-PpoI expression and target plasmids. Total DNA from these cells was digested with NcoI and hybridized with the 1.8 kb BamHI/HindIII fragment of pBC/SacRB as a probe (Lanes 1–5). In lanes 6–10, DNA was also digested with I-SceI or I-PpoI in vitro . The white arrow marks linearized full-length plasmids (lanes 1–5) and plasmids resistant to in vitro endonuclease cleavage. ( c ) Number of Cam/Suc-resistant colonies after bacterial transformation of plasmid DNA isolated from transfected Sua4.0 cells. Co-transfection of the target plasmids together with I-SceI and I-PpoI expression vectors increases the number of colonies. ( d ) Expression of I-SceI and I-PpoI in A. gambiae Sua 4.0 cells. Western blot of transfected cells using anti-hemagglutinin (anti-HA) and anti-α tubulin (left). The I-PpoI-GFP fusion proteins show the expected nuclear localization unlike the Actin 5C-driven GFP control (right).

    Journal: Nucleic Acids Research

    Article Title: Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos

    doi: 10.1093/nar/gkm632

    Figure Lengend Snippet: I-SceI and I-PpoI expression constructs and their activity in A. gambiae cells. ( a ) Maps of HEG expression and target vectors used in the interplasmid activity assay. CMV, cytomegalovirus promoter; Hsp70, Drosophila heatshock protein 70 promoter; eGFP enhanced green fluorescent protein; SacRB , levansucrase gene; X, XbaI; B, BamHI; E, EcoRI; H, HindIII; Xh, XhoI; N, NcoI; A, AscI; S, SalI; Nh, NheI; Kan R , kanamycin resistance cassette; Cam R , chloramphenicol resistance cassette; Tet R , tetracycline resistance cassette; Amp R , ampicilin resistance cassette; NLS, nuclear localization signal. ( b ) Analysis of I-SceI and I-PpoI activity in Sua4.0 cells by Southern blot. Cells were co-transfected with I-SceI or I-PpoI expression and target plasmids. Total DNA from these cells was digested with NcoI and hybridized with the 1.8 kb BamHI/HindIII fragment of pBC/SacRB as a probe (Lanes 1–5). In lanes 6–10, DNA was also digested with I-SceI or I-PpoI in vitro . The white arrow marks linearized full-length plasmids (lanes 1–5) and plasmids resistant to in vitro endonuclease cleavage. ( c ) Number of Cam/Suc-resistant colonies after bacterial transformation of plasmid DNA isolated from transfected Sua4.0 cells. Co-transfection of the target plasmids together with I-SceI and I-PpoI expression vectors increases the number of colonies. ( d ) Expression of I-SceI and I-PpoI in A. gambiae Sua 4.0 cells. Western blot of transfected cells using anti-hemagglutinin (anti-HA) and anti-α tubulin (left). The I-PpoI-GFP fusion proteins show the expected nuclear localization unlike the Actin 5C-driven GFP control (right).

    Article Snippet: In vivo HEG activity assays were performed by lipid-mediated transient transfections (Effecten, Qiagen) of 1–3 × 105 cells with 2 μg/ml (culture volume) recipient plasmid and 4 μg/ml donor plasmid.

    Techniques: Expressing, Construct, Activity Assay, Chick Chorioallantoic Membrane Assay, Southern Blot, Transfection, In Vitro, Electroporation Bacterial Transformation, Plasmid Preparation, Isolation, Cotransfection, Western Blot

    Analysis of I-SceI-induced homologous repair and interplasmid gene conversion events from A. gambiae cells and embryos. ( a ) Reporter plasmid pDR-CMV-GFP. In frame stop codons introduced by I- Sce I recognition site are underlined. ( b ) Fluorescence activated cell sorting (FACS) of transfected cells performed 72 h post-transfection. Two-colour fluorescence analysis of cell line Sua4.0 transfected with pDR-CMV-GFP in the presence and absence of pP[v+,70I-SceI]. A gate using SSC-H (complexity) versus FSC-H (size) was used for cell size analysis (data not shown). The percentage of green fluorescent cells falling below the diagonal for each transfection are indicated (upper panels). The panel at the lower left side shows an overlay of both transfections (pDR-CMV-GFP transfected cells shown in red, pDR-CMV-GFP + pP[v+,70I-SceI] transfected cells shown in green) and the panel on the lower right side shows a histogram of all GFP+ events. FL1-H, green fluorescence; FL2-H, red autofluorescence. ( c ) Detailed maps of pBC/SacRB S1, pSL-SacRB Tet and the predicted product plasmid created by a perfect gene conversion event. ( d ) Resistance properties of pBC/SacRB S1, pSL-SacRB Tet and the predicted product plasmid created by a perfect gene conversion event. ( e ) Restriction analysis of pBC/SacRB S1 and 2 clones isolated from microinjected embryos. X, XbaI; B, BamHI; E, EcoRI; H, HindIII; Xh, XhoI; N, NcoI; A, AscI; S, SalI; Nh, NheI; HL1, Hyperladder 1 (1.5, 2, 2.5, 3, 4, 5, 6 and 10 kb).

    Journal: Nucleic Acids Research

    Article Title: Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos

    doi: 10.1093/nar/gkm632

    Figure Lengend Snippet: Analysis of I-SceI-induced homologous repair and interplasmid gene conversion events from A. gambiae cells and embryos. ( a ) Reporter plasmid pDR-CMV-GFP. In frame stop codons introduced by I- Sce I recognition site are underlined. ( b ) Fluorescence activated cell sorting (FACS) of transfected cells performed 72 h post-transfection. Two-colour fluorescence analysis of cell line Sua4.0 transfected with pDR-CMV-GFP in the presence and absence of pP[v+,70I-SceI]. A gate using SSC-H (complexity) versus FSC-H (size) was used for cell size analysis (data not shown). The percentage of green fluorescent cells falling below the diagonal for each transfection are indicated (upper panels). The panel at the lower left side shows an overlay of both transfections (pDR-CMV-GFP transfected cells shown in red, pDR-CMV-GFP + pP[v+,70I-SceI] transfected cells shown in green) and the panel on the lower right side shows a histogram of all GFP+ events. FL1-H, green fluorescence; FL2-H, red autofluorescence. ( c ) Detailed maps of pBC/SacRB S1, pSL-SacRB Tet and the predicted product plasmid created by a perfect gene conversion event. ( d ) Resistance properties of pBC/SacRB S1, pSL-SacRB Tet and the predicted product plasmid created by a perfect gene conversion event. ( e ) Restriction analysis of pBC/SacRB S1 and 2 clones isolated from microinjected embryos. X, XbaI; B, BamHI; E, EcoRI; H, HindIII; Xh, XhoI; N, NcoI; A, AscI; S, SalI; Nh, NheI; HL1, Hyperladder 1 (1.5, 2, 2.5, 3, 4, 5, 6 and 10 kb).

    Article Snippet: In vivo HEG activity assays were performed by lipid-mediated transient transfections (Effecten, Qiagen) of 1–3 × 105 cells with 2 μg/ml (culture volume) recipient plasmid and 4 μg/ml donor plasmid.

    Techniques: Plasmid Preparation, Fluorescence, FACS, Transfection, Clone Assay, Isolation

    Evaluation of transfection efficiency of B16-F10 cells transfected with use of carrier:pDNA complexes. (a) Percentage of GFP-positive cells transfected with complexes: AP:pGFP at different N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15) and AP-15/DOPE:pGFP, AP-15/DOPE/DMEM:pGFP lipoplexes containing 2.5 μg of lipids/μg of pGFP, analysed by FACS; Ap- significant difference from AP-15 treatment, At- significant difference from Attractene treatment, L- significant difference from Lipofectamine treatment, P- significant difference from PEI treatment, (b) Activity of β-galactosidase in cells transfected with complexes: AP-11:pLacZ, AP-15:pLacZ, PEI:pLacZ, at r = 2.0–2.5 N/P ratio, studied by β-Gal test; *P

    Journal: PLoS ONE

    Article Title: Prenyl Ammonium Salts – New Carriers for Gene Delivery: A B16-F10 Mouse Melanoma Model

    doi: 10.1371/journal.pone.0153633

    Figure Lengend Snippet: Evaluation of transfection efficiency of B16-F10 cells transfected with use of carrier:pDNA complexes. (a) Percentage of GFP-positive cells transfected with complexes: AP:pGFP at different N/P ratios (r = 1.7 for AP-7, AP-8, AP-11, r = 2.0 for AP-15) and AP-15/DOPE:pGFP, AP-15/DOPE/DMEM:pGFP lipoplexes containing 2.5 μg of lipids/μg of pGFP, analysed by FACS; Ap- significant difference from AP-15 treatment, At- significant difference from Attractene treatment, L- significant difference from Lipofectamine treatment, P- significant difference from PEI treatment, (b) Activity of β-galactosidase in cells transfected with complexes: AP-11:pLacZ, AP-15:pLacZ, PEI:pLacZ, at r = 2.0–2.5 N/P ratio, studied by β-Gal test; *P

    Article Snippet: Control experiments with application of nextgeneration lipid reagent Attractene Transfection Reagent (Qiagen) and cationic polymer Satisfection Transfection Reagent (Agilent Technologies) were conducted according to the manufacturer’s recommendations, using the experimentally verified most effective pDNA amounts and reagents volumes.

    Techniques: Transfection, FACS, Activity Assay

    Mutational disruption of NK1R-mediated sequestration of β-arrestin prevents inhibition of F-MOR endocytosis in N2A cells (A) Confocal micrographs of N2A cells co-expressing either full-length HA-NK1R (left panels, red) or truncated HA-NK1R (right panels, red) with β-arrestin2-EGFP (green). Incubation with 10μM SP in cells expressing the truncated HA-NK1R (355x) resulted in less co-localization with receptors in endosomal clusters and higher cytoplasmic β-arrestin2-EGFP distribution (left column), compared to β-arrestin2-EGFP distribution in the wild type HA-NK1R under the same conditions (right column). Scale bar, 10μm. (B) In N2A cells co-expressing F-MOR (red) and HA-NK1 355x receptors (green), both morphine and DAMGO were able to drive internalization of F-MORs even in the presence of substance P. Scale bar, 10μm. (C) Quantification using ratiometric staining of F-MORs in N2A cells co-expressing HA-NK1 355x revealed that co-incubation with SP produced comparable levels of F-MOR internalization compared to treatment with either MS or DG alone. Bar graphs represent mean internalization determined from ~50 cell bodies selected at random in the N2A cultures and averaged over four independent transfections. (D) Biochemical assay of F-MOR internalization by surface biotinylation confirmed that F-MOR internalization induced by either MS or DG (lanes 3 and 5) was not significantly inhibited by co-application of SP (lanes 4 and 6; n= 3 independent experiments).

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Neurokinin1 receptors regulate morphine-induced endocytosis and desensitization of mu opioid receptors in CNS neurons

    doi: 10.1523/JNEUROSCI.4315-08.2009

    Figure Lengend Snippet: Mutational disruption of NK1R-mediated sequestration of β-arrestin prevents inhibition of F-MOR endocytosis in N2A cells (A) Confocal micrographs of N2A cells co-expressing either full-length HA-NK1R (left panels, red) or truncated HA-NK1R (right panels, red) with β-arrestin2-EGFP (green). Incubation with 10μM SP in cells expressing the truncated HA-NK1R (355x) resulted in less co-localization with receptors in endosomal clusters and higher cytoplasmic β-arrestin2-EGFP distribution (left column), compared to β-arrestin2-EGFP distribution in the wild type HA-NK1R under the same conditions (right column). Scale bar, 10μm. (B) In N2A cells co-expressing F-MOR (red) and HA-NK1 355x receptors (green), both morphine and DAMGO were able to drive internalization of F-MORs even in the presence of substance P. Scale bar, 10μm. (C) Quantification using ratiometric staining of F-MORs in N2A cells co-expressing HA-NK1 355x revealed that co-incubation with SP produced comparable levels of F-MOR internalization compared to treatment with either MS or DG alone. Bar graphs represent mean internalization determined from ~50 cell bodies selected at random in the N2A cultures and averaged over four independent transfections. (D) Biochemical assay of F-MOR internalization by surface biotinylation confirmed that F-MOR internalization induced by either MS or DG (lanes 3 and 5) was not significantly inhibited by co-application of SP (lanes 4 and 6; n= 3 independent experiments).

    Article Snippet: Transfections were performed using a cationic lipid transfection reagent (Effectene; Qiagen, Hilden, Germany).

    Techniques: Inhibition, Expressing, Incubation, Staining, Produced, Mass Spectrometry, Transfection

    Heterologous regulation of F-MOR endocytosis in mouse neuroblastoma (N2A) cells (A) Dual-labeled confocal fluorescence micrographs show F-MOR (red) and HA-NK1R (green) transiently expressed in N2A cells. In untreated cells, both F-MOR and HA-NK1R showed a plasma membrane distribution, with minimal labeled receptors present internally in the cell (top row). Incubation with either 10μM MS or 10μM SP for 30 minutes resulted in a selective increase in either labeled F-MORs (second row, first panel) or HA-NK1Rs (third row, middle panel), respectively, present internally in the cell. Simultaneous incubation with MS and SP produced visibly less F-MOR internalization compared to incubation with MS alone (bottom row, first panel). Scale bar, 10μm. (B) Quantification of ratiometric staining in N2A cells show inhibition of F-MOR internalization in response to both MS and DG when SP is also present. Bar graphs represent mean internalization determined from ~50 cell bodies selected at random in N2A cultures and averaged over five independent transfections (**p

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Neurokinin1 receptors regulate morphine-induced endocytosis and desensitization of mu opioid receptors in CNS neurons

    doi: 10.1523/JNEUROSCI.4315-08.2009

    Figure Lengend Snippet: Heterologous regulation of F-MOR endocytosis in mouse neuroblastoma (N2A) cells (A) Dual-labeled confocal fluorescence micrographs show F-MOR (red) and HA-NK1R (green) transiently expressed in N2A cells. In untreated cells, both F-MOR and HA-NK1R showed a plasma membrane distribution, with minimal labeled receptors present internally in the cell (top row). Incubation with either 10μM MS or 10μM SP for 30 minutes resulted in a selective increase in either labeled F-MORs (second row, first panel) or HA-NK1Rs (third row, middle panel), respectively, present internally in the cell. Simultaneous incubation with MS and SP produced visibly less F-MOR internalization compared to incubation with MS alone (bottom row, first panel). Scale bar, 10μm. (B) Quantification of ratiometric staining in N2A cells show inhibition of F-MOR internalization in response to both MS and DG when SP is also present. Bar graphs represent mean internalization determined from ~50 cell bodies selected at random in N2A cultures and averaged over five independent transfections (**p

    Article Snippet: Transfections were performed using a cationic lipid transfection reagent (Effectene; Qiagen, Hilden, Germany).

    Techniques: Labeling, Fluorescence, Incubation, Mass Spectrometry, Produced, Staining, Inhibition, Transfection

    Comparative transfection efficiency of FuGENE HD (FuGENE), X-tremeGENE (X-treme), SuperFect (Super), Lipofectamine 2000 (L-2000), Lipofectamine RNAiMAX (L-iMAX), INT, and mTat/PEI/INT using siRNA targeting β - actin in HSC-3 cells. β - actin mRNA was measured by QRT-PCR and then % remaining β - actin mRNA expression was calculated based on control as 100% ( a ). Cell viability of each of the reagent groups in HaCaT cells were evaluated by MTT assay under the same siRNA and transfection reagent volume ( b ). * p

    Journal: Journal of Nanobiotechnology

    Article Title: Efficient siRNA delivery and gene silencing using a lipopolypeptide hybrid vector mediated by a caveolae-mediated and temperature-dependent endocytic pathway

    doi: 10.1186/s12951-019-0444-8

    Figure Lengend Snippet: Comparative transfection efficiency of FuGENE HD (FuGENE), X-tremeGENE (X-treme), SuperFect (Super), Lipofectamine 2000 (L-2000), Lipofectamine RNAiMAX (L-iMAX), INT, and mTat/PEI/INT using siRNA targeting β - actin in HSC-3 cells. β - actin mRNA was measured by QRT-PCR and then % remaining β - actin mRNA expression was calculated based on control as 100% ( a ). Cell viability of each of the reagent groups in HaCaT cells were evaluated by MTT assay under the same siRNA and transfection reagent volume ( b ). * p

    Article Snippet: Non-viral vectors To compare transfection efficiency of siRNA targeting β -actin (Santa Cruz Biotechnology, Inc. Dallas, TX) with several commercial reagents, Lipofectamine 2000 (Invitrogen, Carlsbad, CA), which is a cationic lipid-based transfection reagent, FuGENE HD (Promega Corp., Fitchburg, WI), which is a blend of lipids and other components, SuperFect (Qiagen, Valencia, CA), which is an activated dendrimer-based reagent, Lipofectamine RNAiMAX (Invitrogen), which is a cationic lipid-based transfection reagent, X-tremeGENE (Roche, Switzerland), which is a blend of lipids and other components, and INT (Polyplus-transfection, France) were used.

    Techniques: Transfection, Quantitative RT-PCR, Expressing, MTT Assay