transfection lipid Search Results


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  • 99
    Thermo Fisher lipid transfections
    Activation of FXN expression and comparison to wild-type cells. ( A ) Time course showing activation of FXN mRNA expression starting 48 h <t>post-transfection</t> in FRDA patient-derived NPCs F4259 (5 µM, n = 2, Optimization 4). ( B ) Expression levels of FXN mRNA in FRDA patient-derived NPCs F4259 and wild-type NPCs C7522 ( n = 4). ( C ) Activation in FRDA patient-derived NPCs F4259. ( D ) Effect of adding anti-AAG ASOs to wild-type NPCs C7522. Relative FXN mRNA levels were measured by RT-qPCR when transfected with oligonucleotides using Optimization 4 ( n = 4) post 72 h transfection. EP(−) is non-oligo treatment, no electroporation control. All data are presented as ±STDEV. (*) P
    Lipid Transfections, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    SignaGen lipid transfection
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipid Transfection, supplied by SignaGen, used in various techniques. Bioz Stars score: 88/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Bio-Rad lipid transfection
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipid Transfection, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Mirus Bio lipid mediated transfection
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipid Mediated Transfection, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 93/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen lipid transfection reagent
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipid Transfection Reagent, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen superfect lipid transfection reagent
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Superfect Lipid Transfection Reagent, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen hiperfect lipid transfection reagent
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Hiperfect Lipid Transfection Reagent, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher oligofectamine lipid transfection reagent
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Oligofectamine Lipid Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Stratagene genejammer lipid transfection reagent
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Genejammer Lipid Transfection Reagent, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Polypus Transfection lipidic transfection reagent interferin
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipidic Transfection Reagent Interferin, supplied by Polypus Transfection, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Polypus Transfection lipidic transfection agent interferin
    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h <t>post-transfection,</t> cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.
    Lipidic Transfection Agent Interferin, supplied by Polypus Transfection, used in various techniques. Bioz Stars score: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher lipid mediated transfection
    a2V inhibition increases Notch signaling in TNBC A–D. TNBC cell line MDA-MB-231 was transfected with siRNA oligonucleotides against a2V or Notch1 along with scrambled control siRNA. Cells were harvested 48 hrs after <t>transfection.</t> Fold change in mRNA expression levels of (A) Notch receptors, (B) Notch Ligands and (C) Notch target genes is shown by qRT-PCR performed on the Notch signaling PCR array. Prior to fold-change calculation, the values were normalized to signal generated from endogenous control 18srRNA. (D) Protein level of Notch1 intracellular domain (N1ICD) following a2V gene silencing is shown by western blot analysis. β actin was used as loading control. E–I. MDA-MB-231 cells were treated with Vehicle Control (DMSO), Bafilomycin A1 (Baf A1 – 0.1 or 0.5 μM) or Gamma Secretase Inhibitor (GSI – 2 μM) for 24 hrs. (E) Protein level of Notch1 intracellular domain (N1ICD) following treatment with Baf A1 or GSI is shown by western blot. β actin was used as loading control. (F) Gene expression expression levels of Hes1 relative to endogenous control 18srRNA is shown. (G) Hes1 protein expression is shown by immunofluorescence (H and I). Independently, MDA-MB-231 and MDA-MB-468 were transfected with a RBP-j Notch reporter construct and then treated with Vehicle control, 0.5 μM BafA1 or 2 μM GSI for 24 hrs. Notch reporter levels in (H) MDA-MB-231 and (I) MDA-MB-468 as measured by luciferase assay. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 compared to control. RBPj: Recombinant Binding Protein Suppressor of Hairless.
    Lipid Mediated Transfection, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 224 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    VennNova novafector lipid transfection reagent
    a2V inhibition increases Notch signaling in TNBC A–D. TNBC cell line MDA-MB-231 was transfected with siRNA oligonucleotides against a2V or Notch1 along with scrambled control siRNA. Cells were harvested 48 hrs after <t>transfection.</t> Fold change in mRNA expression levels of (A) Notch receptors, (B) Notch Ligands and (C) Notch target genes is shown by qRT-PCR performed on the Notch signaling PCR array. Prior to fold-change calculation, the values were normalized to signal generated from endogenous control 18srRNA. (D) Protein level of Notch1 intracellular domain (N1ICD) following a2V gene silencing is shown by western blot analysis. β actin was used as loading control. E–I. MDA-MB-231 cells were treated with Vehicle Control (DMSO), Bafilomycin A1 (Baf A1 – 0.1 or 0.5 μM) or Gamma Secretase Inhibitor (GSI – 2 μM) for 24 hrs. (E) Protein level of Notch1 intracellular domain (N1ICD) following treatment with Baf A1 or GSI is shown by western blot. β actin was used as loading control. (F) Gene expression expression levels of Hes1 relative to endogenous control 18srRNA is shown. (G) Hes1 protein expression is shown by immunofluorescence (H and I). Independently, MDA-MB-231 and MDA-MB-468 were transfected with a RBP-j Notch reporter construct and then treated with Vehicle control, 0.5 μM BafA1 or 2 μM GSI for 24 hrs. Notch reporter levels in (H) MDA-MB-231 and (I) MDA-MB-468 as measured by luciferase assay. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 compared to control. RBPj: Recombinant Binding Protein Suppressor of Hairless.
    Novafector Lipid Transfection Reagent, supplied by VennNova, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher 293fectin lipid transfection
    a2V inhibition increases Notch signaling in TNBC A–D. TNBC cell line MDA-MB-231 was transfected with siRNA oligonucleotides against a2V or Notch1 along with scrambled control siRNA. Cells were harvested 48 hrs after <t>transfection.</t> Fold change in mRNA expression levels of (A) Notch receptors, (B) Notch Ligands and (C) Notch target genes is shown by qRT-PCR performed on the Notch signaling PCR array. Prior to fold-change calculation, the values were normalized to signal generated from endogenous control 18srRNA. (D) Protein level of Notch1 intracellular domain (N1ICD) following a2V gene silencing is shown by western blot analysis. β actin was used as loading control. E–I. MDA-MB-231 cells were treated with Vehicle Control (DMSO), Bafilomycin A1 (Baf A1 – 0.1 or 0.5 μM) or Gamma Secretase Inhibitor (GSI – 2 μM) for 24 hrs. (E) Protein level of Notch1 intracellular domain (N1ICD) following treatment with Baf A1 or GSI is shown by western blot. β actin was used as loading control. (F) Gene expression expression levels of Hes1 relative to endogenous control 18srRNA is shown. (G) Hes1 protein expression is shown by immunofluorescence (H and I). Independently, MDA-MB-231 and MDA-MB-468 were transfected with a RBP-j Notch reporter construct and then treated with Vehicle control, 0.5 μM BafA1 or 2 μM GSI for 24 hrs. Notch reporter levels in (H) MDA-MB-231 and (I) MDA-MB-468 as measured by luciferase assay. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 compared to control. RBPj: Recombinant Binding Protein Suppressor of Hairless.
    293fectin Lipid Transfection, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Mirus Bio lipid based transfection reagent
    RNAi is active in WNV infected cells . RNAi of influenza M2 gene in cells that replicate WNV RNA. Huh7.5, Huh7.5-Rep, and WNV infected Huh7.5 cells (8 hours post infection) were transfected with pCM2 and Cap or M2 siRNA as described in Materials and Methods. 24 hours later cells were processed by flow cytometry for M2 expression using antibody 14C2. The percentage of M2 inhibition was calculated according to the following formula: (1 – (% M2 expression of siRNA-transfected cells / % M2 expression in cells transfected with <t>transfection</t> vehicle only) × 100). The results are an average of three independent experiments and error bars indicate standard error of the mean.
    Lipid Based Transfection Reagent, supplied by Mirus Bio, used in various techniques. Bioz Stars score: 88/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega cationic lipid transfection reagent transfast
    RNAi is active in WNV infected cells . RNAi of influenza M2 gene in cells that replicate WNV RNA. Huh7.5, Huh7.5-Rep, and WNV infected Huh7.5 cells (8 hours post infection) were transfected with pCM2 and Cap or M2 siRNA as described in Materials and Methods. 24 hours later cells were processed by flow cytometry for M2 expression using antibody 14C2. The percentage of M2 inhibition was calculated according to the following formula: (1 – (% M2 expression of siRNA-transfected cells / % M2 expression in cells transfected with <t>transfection</t> vehicle only) × 100). The results are an average of three independent experiments and error bars indicate standard error of the mean.
    Cationic Lipid Transfection Reagent Transfast, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Activation of FXN expression and comparison to wild-type cells. ( A ) Time course showing activation of FXN mRNA expression starting 48 h post-transfection in FRDA patient-derived NPCs F4259 (5 µM, n = 2, Optimization 4). ( B ) Expression levels of FXN mRNA in FRDA patient-derived NPCs F4259 and wild-type NPCs C7522 ( n = 4). ( C ) Activation in FRDA patient-derived NPCs F4259. ( D ) Effect of adding anti-AAG ASOs to wild-type NPCs C7522. Relative FXN mRNA levels were measured by RT-qPCR when transfected with oligonucleotides using Optimization 4 ( n = 4) post 72 h transfection. EP(−) is non-oligo treatment, no electroporation control. All data are presented as ±STDEV. (*) P

    Journal: RNA

    Article Title: Efficient electroporation of neuronal cells using synthetic oligonucleotides: identifying duplex RNA and antisense oligonucleotide activators of human frataxin expression

    doi: 10.1261/rna.071290.119

    Figure Lengend Snippet: Activation of FXN expression and comparison to wild-type cells. ( A ) Time course showing activation of FXN mRNA expression starting 48 h post-transfection in FRDA patient-derived NPCs F4259 (5 µM, n = 2, Optimization 4). ( B ) Expression levels of FXN mRNA in FRDA patient-derived NPCs F4259 and wild-type NPCs C7522 ( n = 4). ( C ) Activation in FRDA patient-derived NPCs F4259. ( D ) Effect of adding anti-AAG ASOs to wild-type NPCs C7522. Relative FXN mRNA levels were measured by RT-qPCR when transfected with oligonucleotides using Optimization 4 ( n = 4) post 72 h transfection. EP(−) is non-oligo treatment, no electroporation control. All data are presented as ±STDEV. (*) P

    Article Snippet: Lipid transfections were performed using Lipofectamine stem transfection reagent (Invitrogen) following reported protocols ( ; ).

    Techniques: Activation Assay, Expressing, Transfection, Derivative Assay, Quantitative RT-PCR, Electroporation

    Identifying an electroporation protocol using iXCells human motor neurons (HMNs). ( A, left ) Relative MALAT-1 RNA levels measured by q RT-PCR when transfected with control gapmer ctrl-gap (1 µM) and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 and 6 ( n = 2) post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2) using 1× trypsin as detachment reagent. ( B, left ) Relative MALAT1 RNA levels measured by q RT-PCR when transfected with control gapmer ctrl-gap (1 µM) and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 2) compared with Lipofectamine stem transfection reagent or gymnotic delivery post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2) using accutase + 10 µM Y27632. EP(−) is non-oligo treatment, no electroporation control. All data are presented as ±STDEV.

    Journal: RNA

    Article Title: Efficient electroporation of neuronal cells using synthetic oligonucleotides: identifying duplex RNA and antisense oligonucleotide activators of human frataxin expression

    doi: 10.1261/rna.071290.119

    Figure Lengend Snippet: Identifying an electroporation protocol using iXCells human motor neurons (HMNs). ( A, left ) Relative MALAT-1 RNA levels measured by q RT-PCR when transfected with control gapmer ctrl-gap (1 µM) and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 and 6 ( n = 2) post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2) using 1× trypsin as detachment reagent. ( B, left ) Relative MALAT1 RNA levels measured by q RT-PCR when transfected with control gapmer ctrl-gap (1 µM) and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 2) compared with Lipofectamine stem transfection reagent or gymnotic delivery post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2) using accutase + 10 µM Y27632. EP(−) is non-oligo treatment, no electroporation control. All data are presented as ±STDEV.

    Article Snippet: Lipid transfections were performed using Lipofectamine stem transfection reagent (Invitrogen) following reported protocols ( ; ).

    Techniques: Electroporation, Reverse Transcription Polymerase Chain Reaction, Transfection, Staining

    Activation of FXN protein expression, western analysis. ( A ) FXN protein in FRDA patient-derived NPCs F4259 and wild-type NPCs C7522 ( n = 3) with different confluency. ( B ) FXN protein expression in FRDA patient-derived NPCs F4259 when transfected with oligonucleotides (5 µM) using Optimization 4 ( n = 3) post 96 h transfection. All data are presented as ±STDEV. (*) P

    Journal: RNA

    Article Title: Efficient electroporation of neuronal cells using synthetic oligonucleotides: identifying duplex RNA and antisense oligonucleotide activators of human frataxin expression

    doi: 10.1261/rna.071290.119

    Figure Lengend Snippet: Activation of FXN protein expression, western analysis. ( A ) FXN protein in FRDA patient-derived NPCs F4259 and wild-type NPCs C7522 ( n = 3) with different confluency. ( B ) FXN protein expression in FRDA patient-derived NPCs F4259 when transfected with oligonucleotides (5 µM) using Optimization 4 ( n = 3) post 96 h transfection. All data are presented as ±STDEV. (*) P

    Article Snippet: Lipid transfections were performed using Lipofectamine stem transfection reagent (Invitrogen) following reported protocols ( ; ).

    Techniques: Activation Assay, Expressing, Western Blot, Derivative Assay, Transfection

    Identifying an electroporation protocol for FRDA patient-derived F4259 iPSC-NPCs. ( A, left ) Relative MALAT1 RNA levels measured by RT-qPCR when transfected with control gapmer ctrl-gap and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 2) compared with Lipofectamine stem transfection reagent and gymnotic delivery post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2). ( B, left ) Relative MALAT1 RNA levels measured by qRT-PCR when transfected with control gapmer ctrl-gap and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 4) post 48 h transfection. ( Right ) Cell viability post 48 h transfection measured by trypan blue staining ( n = 4). ( C ) Time course experiments showing inhibition of MALAT1 RNA expression (1 µM, n = 2, Optimization 4). EP(−) is non-oligo treatment, no electroporation control. All data are presented as ± STDEV.

    Journal: RNA

    Article Title: Efficient electroporation of neuronal cells using synthetic oligonucleotides: identifying duplex RNA and antisense oligonucleotide activators of human frataxin expression

    doi: 10.1261/rna.071290.119

    Figure Lengend Snippet: Identifying an electroporation protocol for FRDA patient-derived F4259 iPSC-NPCs. ( A, left ) Relative MALAT1 RNA levels measured by RT-qPCR when transfected with control gapmer ctrl-gap and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 2) compared with Lipofectamine stem transfection reagent and gymnotic delivery post 24 h transfection. ( Right ) Cell viability post 24 h transfection measured by trypan blue staining ( n = 2). ( B, left ) Relative MALAT1 RNA levels measured by qRT-PCR when transfected with control gapmer ctrl-gap and anti-MALAT1 gapmer anti-MALAT1 using Optimization 4 ( n = 4) post 48 h transfection. ( Right ) Cell viability post 48 h transfection measured by trypan blue staining ( n = 4). ( C ) Time course experiments showing inhibition of MALAT1 RNA expression (1 µM, n = 2, Optimization 4). EP(−) is non-oligo treatment, no electroporation control. All data are presented as ± STDEV.

    Article Snippet: Lipid transfections were performed using Lipofectamine stem transfection reagent (Invitrogen) following reported protocols ( ; ).

    Techniques: Electroporation, Derivative Assay, Quantitative RT-PCR, Transfection, Staining, Inhibition, RNA Expression

    SPC25 knockdown inhibits the proliferation of PC-3 cells. (A) Expression of SPC25 mRNA following transfection with the indicated shRNAs in PC-3 cells. (B) An MTT assay demonstrated that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (C) The Celigo ® system revealed that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (D) Cell number of shCtrl or shSPC25 group in each day was calculated using the Celigo ® system. P

    Journal: Oncology Letters

    Article Title: Knockdown of spindle pole body component 25 homolog inhibits cell proliferation and cycle progression in prostate cancer

    doi: 10.3892/ol.2018.8003

    Figure Lengend Snippet: SPC25 knockdown inhibits the proliferation of PC-3 cells. (A) Expression of SPC25 mRNA following transfection with the indicated shRNAs in PC-3 cells. (B) An MTT assay demonstrated that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (C) The Celigo ® system revealed that knockdown of SPC25 inhibited cell proliferation in PC-3 cells. (D) Cell number of shCtrl or shSPC25 group in each day was calculated using the Celigo ® system. P

    Article Snippet: Opti-modified Eagle's medium (Opti-MEM) which was ideal for use during cationic lipid transfections especially Lipofectamine 2000 transfection reagents was purchased from Thermo Fisher Scientific, Inc (cat. no. 31985062).

    Techniques: Expressing, Transfection, MTT Assay

    iRGD nanoparticles mediate TNF-α silencing by RNAi in vitro . ( A ) Nanoparticle delivery of TNFα siRNA to HEI-193 vestibular schwannoma cells. Relative TNFα gene expression quantified by qRT-PCR in cells treated with various formulations, in the absence of lipopolysaccharide (LPS) stimulation. Medium, untreated controls. siTNFa / scr, control nanoparticles carrying TNFα siRNA. si GFP / iRGD, iRGD nanoparticles carrying GFP siRNA. si TNFα / Lipofect, Lipofectamine RNAiMAX® complexed with TNFα siRNA as a positive transfection control. si TNF α / iRGD, iRGD nanoparticles carrying TNFα siRNA. Error bars represent standard deviation. N = 6 biological replicates with independently prepared nanoparticles. *** P

    Journal: Scientific Reports

    Article Title: Tumor-Penetrating Delivery of siRNA against TNFα to Human Vestibular Schwannomas

    doi: 10.1038/s41598-017-13032-9

    Figure Lengend Snippet: iRGD nanoparticles mediate TNF-α silencing by RNAi in vitro . ( A ) Nanoparticle delivery of TNFα siRNA to HEI-193 vestibular schwannoma cells. Relative TNFα gene expression quantified by qRT-PCR in cells treated with various formulations, in the absence of lipopolysaccharide (LPS) stimulation. Medium, untreated controls. siTNFa / scr, control nanoparticles carrying TNFα siRNA. si GFP / iRGD, iRGD nanoparticles carrying GFP siRNA. si TNFα / Lipofect, Lipofectamine RNAiMAX® complexed with TNFα siRNA as a positive transfection control. si TNF α / iRGD, iRGD nanoparticles carrying TNFα siRNA. Error bars represent standard deviation. N = 6 biological replicates with independently prepared nanoparticles. *** P

    Article Snippet: A commercial lipid transfection reagent, Lipofectamine RNAiMax (Invitrogen), was used as a positive control according to manufacturer’s instructions.

    Techniques: In Vitro, Expressing, Quantitative RT-PCR, Transfection, Standard Deviation

    Expression of ORAI homologs in human airway smooth muscle (HASM) and siRNA-targeted knockdown of ORAI homologs. ( A ) mRNA expression of ORAI1, 2, and 3 in HASM cells using reverse transcriptase–polymerase chain reaction (RT-PCR) (O1: ORAI1, O2: ORAI2, O3: ORAI3). PCR products were sequenced to confirm expression. No products were detected in the RT samples (i.e., RNA samples that have not been reverse transcribed) to confirm no genomic DNA contamination. ( B ) Western blot analysis of ORAI1 protein expression after ORAI1 siRNA transfection; expression of smooth muscle α-actin was assessed as a control for equal protein loading. ( C ) siRNA-mediated knockdown of ORAI1, 2, and 3 mRNA assessed by real-time, quantitative PCR; the effects of individual siRNAs on expression of all ORAI homologs were assessed. The level of ORAI mRNA expression in HASM cells transfected with siRNA was compared relative to an untreated control, which was set to 100%. Results are expressed as the mean ± SEM, cDNA samples were tested in triplicate and the graph represents data from three separate transfections.

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: ORAI and Store-Operated Calcium Influx in Human Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2007-0395OC

    Figure Lengend Snippet: Expression of ORAI homologs in human airway smooth muscle (HASM) and siRNA-targeted knockdown of ORAI homologs. ( A ) mRNA expression of ORAI1, 2, and 3 in HASM cells using reverse transcriptase–polymerase chain reaction (RT-PCR) (O1: ORAI1, O2: ORAI2, O3: ORAI3). PCR products were sequenced to confirm expression. No products were detected in the RT samples (i.e., RNA samples that have not been reverse transcribed) to confirm no genomic DNA contamination. ( B ) Western blot analysis of ORAI1 protein expression after ORAI1 siRNA transfection; expression of smooth muscle α-actin was assessed as a control for equal protein loading. ( C ) siRNA-mediated knockdown of ORAI1, 2, and 3 mRNA assessed by real-time, quantitative PCR; the effects of individual siRNAs on expression of all ORAI homologs were assessed. The level of ORAI mRNA expression in HASM cells transfected with siRNA was compared relative to an untreated control, which was set to 100%. Results are expressed as the mean ± SEM, cDNA samples were tested in triplicate and the graph represents data from three separate transfections.

    Article Snippet: Delivery of siRNA to the cells was achieved through use of lipid-based transfection by Lipofectamine 2000 (Invitrogen).

    Techniques: Expressing, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Real-time Polymerase Chain Reaction

    Huntingtin is required for normal cell heat shock stress response and mutant huntingtin affects the rate of the nuclear cofilin rod stress response. Temporal imaging in live STHdh Q7/Q7 or STHdh Q111/Q111 cells stably expressing mCerulean–cofilin fusion protein. ( A ) mCerulean–cofilin imaged over time showing nuclear cofilin rod formation in both wild-type and mutant cell lines at 10 and 32 min, respectively (b versus g), following panels show length of times rods exist and time of clearance during maintained heat shock at 42°C (c–e and h–j). ( B ) Comparison graph of average time to nuclear rod formation for stable mCerulean–cofilin wild-type ( N = 15) and mutant ( N = 9) STHdh cells during live cell imaging experiments. * P -value of 0.007. ( C ) Temporal imaging in live STHdh Q7/Q7 cells stably expressing mCerulean–cofilin. Cells were co-transfected with control siRNA or huntingtin-specific siRNA and Block-iT™ Alexa Fluor® red for 72 h prior to experiments. Single-cell visualization of control (a–b) or huntingtin siRNA (h–i) transfection with labeled Block-iT™Alexa Fluor® red. Visualization of STHdh Q7/Q7 cells with mCerulean–cofilin during heat shock, treated with control siRNA (c–g). STHdh Q7/Q7 cells with cofilin–mCerulean during heat shock, treated with siRNA to huntingtin (j–n). ( D ) Comparison graph of average time to cell death in mCerulean–cofilin stable STHdh Q7/Q7 ( N = 13) or STHdh Q111/Q111 ( N = 24) cells imaged live and STHdh Q7/Q7 mCerulean–cofilin cells treated with control siRNA ( N = 5) or huntingtin siRNA ( N = 6) imaged live during heat shock. * P -value of 0.003. Scale bars are 10 µm.

    Journal: Human Molecular Genetics

    Article Title: Mutant huntingtin causes defective actin remodeling during stress: defining a new role for transglutaminase 2 in neurodegenerative disease

    doi: 10.1093/hmg/ddr075

    Figure Lengend Snippet: Huntingtin is required for normal cell heat shock stress response and mutant huntingtin affects the rate of the nuclear cofilin rod stress response. Temporal imaging in live STHdh Q7/Q7 or STHdh Q111/Q111 cells stably expressing mCerulean–cofilin fusion protein. ( A ) mCerulean–cofilin imaged over time showing nuclear cofilin rod formation in both wild-type and mutant cell lines at 10 and 32 min, respectively (b versus g), following panels show length of times rods exist and time of clearance during maintained heat shock at 42°C (c–e and h–j). ( B ) Comparison graph of average time to nuclear rod formation for stable mCerulean–cofilin wild-type ( N = 15) and mutant ( N = 9) STHdh cells during live cell imaging experiments. * P -value of 0.007. ( C ) Temporal imaging in live STHdh Q7/Q7 cells stably expressing mCerulean–cofilin. Cells were co-transfected with control siRNA or huntingtin-specific siRNA and Block-iT™ Alexa Fluor® red for 72 h prior to experiments. Single-cell visualization of control (a–b) or huntingtin siRNA (h–i) transfection with labeled Block-iT™Alexa Fluor® red. Visualization of STHdh Q7/Q7 cells with mCerulean–cofilin during heat shock, treated with control siRNA (c–g). STHdh Q7/Q7 cells with cofilin–mCerulean during heat shock, treated with siRNA to huntingtin (j–n). ( D ) Comparison graph of average time to cell death in mCerulean–cofilin stable STHdh Q7/Q7 ( N = 13) or STHdh Q111/Q111 ( N = 24) cells imaged live and STHdh Q7/Q7 mCerulean–cofilin cells treated with control siRNA ( N = 5) or huntingtin siRNA ( N = 6) imaged live during heat shock. * P -value of 0.003. Scale bars are 10 µm.

    Article Snippet: 120 pmol of RNAi was transfected into 25 mm dishes plated at 50% confluency using the lipid-based transfection reagent, Lipofectamine™ 2000 (Invitrogen) according to the manufacturers protocols.

    Techniques: Mutagenesis, Imaging, Stable Transfection, Expressing, Live Cell Imaging, Transfection, Blocking Assay, Labeling

    Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.

    Journal: Retrovirology

    Article Title: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2

    doi: 10.1186/1742-4690-8-64

    Figure Lengend Snippet: Nef residues important for enhancement of viral replication and cell activation are also important for association of Nef with PAK2 in co-precipitation assays . A . 293T cells were transfected with 0.5 μg pCR3.1 Nef-HA or empty vector, 0.5 μg pCDNA.31 FLAG-PAK2 K278R, and 0.5 μg CDC42 V12 in 6-well plates. 48 h post-transfection, cells were lysed in 1ml 1% NP-40 lysis buffer. 10 μl of anti-FLAG agarose conjugate beads were used to immunoprecipitate FLAG-PAK2 from 1 mg of lysate. Bead bound proteins were eluted in 2X SDS sample buffer, separated on a 12% polyacrylamide gel, and transferred to PVDF. Expression was detected by Western blot.

    Article Snippet: 3.5 μg of pHAGE vector,3.5 μg of the packaging construct pDR89.1 [ ], and 1 μg of pVSV-G were transfected by lipid transfection (LipoD293T, Signagen) into 5.5 × 106 293T cells plated 24 h prior in a 10-cm tissue culture plate.

    Techniques: Activation Assay, Transfection, Plasmid Preparation, Lysis, Expressing, Western Blot

    a2V inhibition increases Notch signaling in TNBC A–D. TNBC cell line MDA-MB-231 was transfected with siRNA oligonucleotides against a2V or Notch1 along with scrambled control siRNA. Cells were harvested 48 hrs after transfection. Fold change in mRNA expression levels of (A) Notch receptors, (B) Notch Ligands and (C) Notch target genes is shown by qRT-PCR performed on the Notch signaling PCR array. Prior to fold-change calculation, the values were normalized to signal generated from endogenous control 18srRNA. (D) Protein level of Notch1 intracellular domain (N1ICD) following a2V gene silencing is shown by western blot analysis. β actin was used as loading control. E–I. MDA-MB-231 cells were treated with Vehicle Control (DMSO), Bafilomycin A1 (Baf A1 – 0.1 or 0.5 μM) or Gamma Secretase Inhibitor (GSI – 2 μM) for 24 hrs. (E) Protein level of Notch1 intracellular domain (N1ICD) following treatment with Baf A1 or GSI is shown by western blot. β actin was used as loading control. (F) Gene expression expression levels of Hes1 relative to endogenous control 18srRNA is shown. (G) Hes1 protein expression is shown by immunofluorescence (H and I). Independently, MDA-MB-231 and MDA-MB-468 were transfected with a RBP-j Notch reporter construct and then treated with Vehicle control, 0.5 μM BafA1 or 2 μM GSI for 24 hrs. Notch reporter levels in (H) MDA-MB-231 and (I) MDA-MB-468 as measured by luciferase assay. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 compared to control. RBPj: Recombinant Binding Protein Suppressor of Hairless.

    Journal: Oncotarget

    Article Title: The Vacuolar ATPase a2-subunit regulates Notch signaling in triple-negative breast cancer cells

    doi:

    Figure Lengend Snippet: a2V inhibition increases Notch signaling in TNBC A–D. TNBC cell line MDA-MB-231 was transfected with siRNA oligonucleotides against a2V or Notch1 along with scrambled control siRNA. Cells were harvested 48 hrs after transfection. Fold change in mRNA expression levels of (A) Notch receptors, (B) Notch Ligands and (C) Notch target genes is shown by qRT-PCR performed on the Notch signaling PCR array. Prior to fold-change calculation, the values were normalized to signal generated from endogenous control 18srRNA. (D) Protein level of Notch1 intracellular domain (N1ICD) following a2V gene silencing is shown by western blot analysis. β actin was used as loading control. E–I. MDA-MB-231 cells were treated with Vehicle Control (DMSO), Bafilomycin A1 (Baf A1 – 0.1 or 0.5 μM) or Gamma Secretase Inhibitor (GSI – 2 μM) for 24 hrs. (E) Protein level of Notch1 intracellular domain (N1ICD) following treatment with Baf A1 or GSI is shown by western blot. β actin was used as loading control. (F) Gene expression expression levels of Hes1 relative to endogenous control 18srRNA is shown. (G) Hes1 protein expression is shown by immunofluorescence (H and I). Independently, MDA-MB-231 and MDA-MB-468 were transfected with a RBP-j Notch reporter construct and then treated with Vehicle control, 0.5 μM BafA1 or 2 μM GSI for 24 hrs. Notch reporter levels in (H) MDA-MB-231 and (I) MDA-MB-468 as measured by luciferase assay. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001 compared to control. RBPj: Recombinant Binding Protein Suppressor of Hairless.

    Article Snippet: Cells were transfected with siRNA (final concentration, 10 nM) using lipid-mediated transfection with Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's instructions.

    Techniques: Inhibition, Multiple Displacement Amplification, Transfection, Expressing, Quantitative RT-PCR, Polymerase Chain Reaction, Generated, Western Blot, Immunofluorescence, Construct, Luciferase, Recombinant, Binding Assay

    a2V-ATPase inhibition enhances Wnt signaling in TNBC A. MDA-MB-231 cells were transfected with scrambled control or a2V siRNA and harvested after 48 hrs of transfection. Fold change in mRNA expression levels of Wnt signaling genes WNT4, β-catenin (CTNNB1), C-MYC and Cyclin D1 (CYCD1) was assessed by qRT PCR. Prior to fold--change calculation, the values were normalized to signal generated from endogenous control 18srRNA. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01 compared to control siRNA. (B and C) TNBC cells were grown on chamber slides and treated with B. vehicle control or C. 0.1 μM Baf A1 for 4 hours. Cells were fixed, permeabilized and processed for immunofluorescence microscopy. Localization of β-catenin (green) is shown. Nucleus was stained with DAPI (blue). Scale bars: 10 μm.

    Journal: Oncotarget

    Article Title: The Vacuolar ATPase a2-subunit regulates Notch signaling in triple-negative breast cancer cells

    doi:

    Figure Lengend Snippet: a2V-ATPase inhibition enhances Wnt signaling in TNBC A. MDA-MB-231 cells were transfected with scrambled control or a2V siRNA and harvested after 48 hrs of transfection. Fold change in mRNA expression levels of Wnt signaling genes WNT4, β-catenin (CTNNB1), C-MYC and Cyclin D1 (CYCD1) was assessed by qRT PCR. Prior to fold--change calculation, the values were normalized to signal generated from endogenous control 18srRNA. Data represent mean ± standard error, n = 4. * P ≤ 0.05, ** P ≤ 0.01 compared to control siRNA. (B and C) TNBC cells were grown on chamber slides and treated with B. vehicle control or C. 0.1 μM Baf A1 for 4 hours. Cells were fixed, permeabilized and processed for immunofluorescence microscopy. Localization of β-catenin (green) is shown. Nucleus was stained with DAPI (blue). Scale bars: 10 μm.

    Article Snippet: Cells were transfected with siRNA (final concentration, 10 nM) using lipid-mediated transfection with Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer's instructions.

    Techniques: Inhibition, Multiple Displacement Amplification, Transfection, Expressing, Quantitative RT-PCR, Generated, Immunofluorescence, Microscopy, Staining

    RNAi is active in WNV infected cells . RNAi of influenza M2 gene in cells that replicate WNV RNA. Huh7.5, Huh7.5-Rep, and WNV infected Huh7.5 cells (8 hours post infection) were transfected with pCM2 and Cap or M2 siRNA as described in Materials and Methods. 24 hours later cells were processed by flow cytometry for M2 expression using antibody 14C2. The percentage of M2 inhibition was calculated according to the following formula: (1 – (% M2 expression of siRNA-transfected cells / % M2 expression in cells transfected with transfection vehicle only) × 100). The results are an average of three independent experiments and error bars indicate standard error of the mean.

    Journal: Virology Journal

    Article Title: Actively replicating West Nile virus is resistant to cytoplasmic delivery of siRNA

    doi: 10.1186/1743-422X-2-53

    Figure Lengend Snippet: RNAi is active in WNV infected cells . RNAi of influenza M2 gene in cells that replicate WNV RNA. Huh7.5, Huh7.5-Rep, and WNV infected Huh7.5 cells (8 hours post infection) were transfected with pCM2 and Cap or M2 siRNA as described in Materials and Methods. 24 hours later cells were processed by flow cytometry for M2 expression using antibody 14C2. The percentage of M2 inhibition was calculated according to the following formula: (1 – (% M2 expression of siRNA-transfected cells / % M2 expression in cells transfected with transfection vehicle only) × 100). The results are an average of three independent experiments and error bars indicate standard error of the mean.

    Article Snippet: Because the lipid-based transfection reagent targets nucleic acids to the cytoplasm (Mirus Corp, personal communication), the presence of additional membranes between the viral RNA and the cytoplasm could prevent the siRNA from reaching the actively replicating WNV RNA complex.

    Techniques: Infection, Transfection, Flow Cytometry, Cytometry, Expressing, Inhibition