sirna transfection medium Search Results


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  • 99
    Thermo Fisher sirna transfection medium lipofectamine rnaimax
    Sirna Transfection Medium Lipofectamine Rnaimax, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 2 article reviews
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    99
    Millipore sirna transfection medium
    Sirna Transfection Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna transfection medium/product/Millipore
    Average 99 stars, based on 14 article reviews
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    sirna transfection medium - by Bioz Stars, 2020-08
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    92
    Santa Cruz Biotechnology sirna transfection medium
    Slug regulation-induced processing of Slug and PUMA. ( A ) The protein fractions of Slug in QBC939, RBE, ICC-9810, and FRH 0201 cells were subjected to Western blot analysis. FRH 0201 exhibited the lowest expression level of Slug and QBC939 exhibited the highest expression level of Slug. ( B ) The protein fractions of Slug, E-cadherin, and PUMA in QBC939 cells after <t>transfection</t> with Slug <t>siRNA</t> for 48 h were subjected to Western blot analysis. Slug expression was barely detectable compared with parental cells (** P
    Sirna Transfection Medium, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1066 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna transfection medium/product/Santa Cruz Biotechnology
    Average 92 stars, based on 1066 article reviews
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    92
    Fisher Scientific sirna transfection medium
    Slug regulation-induced processing of Slug and PUMA. ( A ) The protein fractions of Slug in QBC939, RBE, ICC-9810, and FRH 0201 cells were subjected to Western blot analysis. FRH 0201 exhibited the lowest expression level of Slug and QBC939 exhibited the highest expression level of Slug. ( B ) The protein fractions of Slug, E-cadherin, and PUMA in QBC939 cells after <t>transfection</t> with Slug <t>siRNA</t> for 48 h were subjected to Western blot analysis. Slug expression was barely detectable compared with parental cells (** P
    Sirna Transfection Medium, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sirna transfection medium/product/Fisher Scientific
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    92
    Thermo Fisher sirna transfection medium
    Effects of <t>siRNA</t> BGR1 knockdown on the ATP depletion–induced expression of TNF- α and MCP-1 genes in proximal tubule HK-2 cell culture. After <t>transfection</t> with either BRG1 siRNA (■) or noncomplementary (NC) siRNA ( ), HK-2 cells
    Sirna Transfection Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    sirna transfection medium - by Bioz Stars, 2020-08
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    88
    Santa Cruz Biotechnology shrna plasmid transfection medium
    Effects of <t>siRNA</t> BGR1 knockdown on the ATP depletion–induced expression of TNF- α and MCP-1 genes in proximal tubule HK-2 cell culture. After <t>transfection</t> with either BRG1 siRNA (■) or noncomplementary (NC) siRNA ( ), HK-2 cells
    Shrna Plasmid Transfection Medium, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology sirna transfection medium sc 36868
    CD74 knockdown by CD74 <t>siRNA</t> <t>transfection</t> reagent in CAMA-1 and MDA-MB-231 cells; A) and B) siRNA-mediated knockdown of CD74 expression in CAMA-1 and MDA-MB-231 cells were detected by Western blot; An approximately two to five (siRNA) fold weaker signal of CD74 protein expression is apparent, as compared to the negative control siRNA group normalized to the expression of α-Tubulin; C) and D) Confocal images of CAMA-1 and MDA-MB-231 cells transfected with CD74 siRNA, untreated control and negative control siRNA; Data represent three different experiments
    Sirna Transfection Medium Sc 36868, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology antibiotic free sirna transfection medium
    CD5L serves a cytoprotective function by modulating autophagy. (A-E) To determine the role of autophagy in the anti-apoptotic actions of CD5L, hepatocytes were transfected with <t>siRNA</t> against ATG7 or siRNA-NT. (A) The siRNA-mediated <t>transfection</t> efficiency was demonstrated by reverse transcription-quantitative PCR. Each column represents the mean ± SD of three independent experiments. * P
    Antibiotic Free Sirna Transfection Medium, supplied by Santa Cruz Biotechnology, 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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    92
    Santa Cruz Biotechnology serum free sirna transfection medium
    Tenascin C contributes to NRP-1 associated migration. a. Dual immunofluorescence staining (40× magnification scale bar 10 μm) of NRP-1 and TNC on BT-474 and BT-474 NRP-1 cells indicates their colocalization in the cytoplasm. Treatment of BT-474 NRP-1 cells with TNC targeted <t>siRNA</t> molecules, b , reduced TNC gene expression, c , reduced migratory capacity and d , downregulated NRP-1 and vimentin expression. (TNC protein was not detected on western blot due to the lack of specific antibody for this application.) The gene expression fold change was measured by comparing the basal levels detected in the empty plasmid transfected BT-474 or in the case of the siRNA experiment, to the control siRNA treated BT-474 and normalized to β-Actin and GUSB reference gene expression. Wound healing assay images (panel d, 5× magnification, scale bar 500 μm) taken on day 0 and day 2 after siRNA <t>transfection.</t> Graphs represent the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value
    Serum Free Sirna Transfection Medium, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Horizon Discovery accell sirna transfection medium
    Tenascin C contributes to NRP-1 associated migration. a. Dual immunofluorescence staining (40× magnification scale bar 10 μm) of NRP-1 and TNC on BT-474 and BT-474 NRP-1 cells indicates their colocalization in the cytoplasm. Treatment of BT-474 NRP-1 cells with TNC targeted <t>siRNA</t> molecules, b , reduced TNC gene expression, c , reduced migratory capacity and d , downregulated NRP-1 and vimentin expression. (TNC protein was not detected on western blot due to the lack of specific antibody for this application.) The gene expression fold change was measured by comparing the basal levels detected in the empty plasmid transfected BT-474 or in the case of the siRNA experiment, to the control siRNA treated BT-474 and normalized to β-Actin and GUSB reference gene expression. Wound healing assay images (panel d, 5× magnification, scale bar 500 μm) taken on day 0 and day 2 after siRNA <t>transfection.</t> Graphs represent the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value
    Accell Sirna Transfection Medium, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Santa Cruz Biotechnology syk sirna transfection medium
    Kinases Lyn and <t>Syk</t> selectively regulate Ras isoform’s activation through Ras-GEFs. a CD40 activates the upstream kinases Lyn and Syk in a dose-dependent manner, with lower doses activating Syk and higher doses activating Lyn. b Immunoblot analysis of total Ras and activated Ras isoforms in the lysates of un-transfected or Syk- or Lyn-specific <t>siRNA</t> transfected, anti-CD40 antibody (3 μg/ml) treated P388D1 cells. c Immunoblot analyses of phospho-tyrosine (p-tyr-Vav)), and Vav translocated Sos-1/2 (Tr-Sos-1/2), translocated Ras-GRP (Tr-RasGRP), CD71, syk, lyn and β-actin in the lysates of untreated or Syk siRNA or Lyn siRNA or anti-CD40 antibody (3 μg/ml) treated P388D1 cells. d Immunoblot analysis of phospho-tyrosine (p-tyr Vav) and Vav translocated Sos-1/2, translocated Ras GRP, CD71, Lyn and phospho-Lyn, Syk and phospho-Syk (p-syk) in the lysates of untreated or Syk inhibitor (Syk Inh, 3 μM; Calbiochem, San Diego, CA) or PP-1 (340 nM; BIOMOL International, PA) treated and/or anti-CD40 antibody (3 μg/ml) treated macrophages. e Analysis of the association of Syk or Lyn with Sos-1/2, Vav, and Ras-GRP in the co-immunoprecipitates from the lysates of anti-CD40 antibody (1 μg/ml, 3 μg/ml and 6 μg/ml) treated macrophages.
    Syk Sirna Transfection Medium, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology shrna transfection medium santa cruz biotech sc 108062
    Kinases Lyn and <t>Syk</t> selectively regulate Ras isoform’s activation through Ras-GEFs. a CD40 activates the upstream kinases Lyn and Syk in a dose-dependent manner, with lower doses activating Syk and higher doses activating Lyn. b Immunoblot analysis of total Ras and activated Ras isoforms in the lysates of un-transfected or Syk- or Lyn-specific <t>siRNA</t> transfected, anti-CD40 antibody (3 μg/ml) treated P388D1 cells. c Immunoblot analyses of phospho-tyrosine (p-tyr-Vav)), and Vav translocated Sos-1/2 (Tr-Sos-1/2), translocated Ras-GRP (Tr-RasGRP), CD71, syk, lyn and β-actin in the lysates of untreated or Syk siRNA or Lyn siRNA or anti-CD40 antibody (3 μg/ml) treated P388D1 cells. d Immunoblot analysis of phospho-tyrosine (p-tyr Vav) and Vav translocated Sos-1/2, translocated Ras GRP, CD71, Lyn and phospho-Lyn, Syk and phospho-Syk (p-syk) in the lysates of untreated or Syk inhibitor (Syk Inh, 3 μM; Calbiochem, San Diego, CA) or PP-1 (340 nM; BIOMOL International, PA) treated and/or anti-CD40 antibody (3 μg/ml) treated macrophages. e Analysis of the association of Syk or Lyn with Sos-1/2, Vav, and Ras-GRP in the co-immunoprecipitates from the lysates of anti-CD40 antibody (1 μg/ml, 3 μg/ml and 6 μg/ml) treated macrophages.
    Shrna Transfection Medium Santa Cruz Biotech Sc 108062, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/shrna transfection medium santa cruz biotech sc 108062/product/Santa Cruz Biotechnology
    Average 93 stars, based on 14 article reviews
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    93
    Polyplus Transfection jetprime sirna transfection protocol
    Kinases Lyn and <t>Syk</t> selectively regulate Ras isoform’s activation through Ras-GEFs. a CD40 activates the upstream kinases Lyn and Syk in a dose-dependent manner, with lower doses activating Syk and higher doses activating Lyn. b Immunoblot analysis of total Ras and activated Ras isoforms in the lysates of un-transfected or Syk- or Lyn-specific <t>siRNA</t> transfected, anti-CD40 antibody (3 μg/ml) treated P388D1 cells. c Immunoblot analyses of phospho-tyrosine (p-tyr-Vav)), and Vav translocated Sos-1/2 (Tr-Sos-1/2), translocated Ras-GRP (Tr-RasGRP), CD71, syk, lyn and β-actin in the lysates of untreated or Syk siRNA or Lyn siRNA or anti-CD40 antibody (3 μg/ml) treated P388D1 cells. d Immunoblot analysis of phospho-tyrosine (p-tyr Vav) and Vav translocated Sos-1/2, translocated Ras GRP, CD71, Lyn and phospho-Lyn, Syk and phospho-Syk (p-syk) in the lysates of untreated or Syk inhibitor (Syk Inh, 3 μM; Calbiochem, San Diego, CA) or PP-1 (340 nM; BIOMOL International, PA) treated and/or anti-CD40 antibody (3 μg/ml) treated macrophages. e Analysis of the association of Syk or Lyn with Sos-1/2, Vav, and Ras-GRP in the co-immunoprecipitates from the lysates of anti-CD40 antibody (1 μg/ml, 3 μg/ml and 6 μg/ml) treated macrophages.
    Jetprime Sirna Transfection Protocol, supplied by Polyplus Transfection, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    jetprime sirna transfection protocol - by Bioz Stars, 2020-08
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    99
    Thermo Fisher medium throughput rodent mirna
    Unsupervised hierarchical clustering of miRNAs reveals relationship of groups. Experimental groups were largely recapitulated by unsupervised hierarchical clustering of all results from <t>OpenArray</t> <t>miRNA</t> profiling. Clustering was performed by Pearson correlation
    Medium Throughput Rodent Mirna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher transfection medium
    knock-down of endogenous CHMP2B in hippocampal neurones. A: to verify the efficacy of the shRNA-encoding plasmid, the indicated plasmids were transfected in rodent (BHK) cells and equal protein amounts of transfected cell lysates were analysed by Western immunoblotting with anti-CHMP2B antibody, or with anti-actin as a control. Lane 1: empty pSuper-mCherry plasmid; lane 2: shRNA-expressing plasmid; lane 3: shRNA-expressing plasmid cotransfected with vector encoding CHMP2B* (native CHMP2B cDNA with silent mutations at the siRNA target site). Note that the CHMP2B protein remaining after <t>transfection</t> of the shRNA plasmid largely originates from non-transfected cells in the culture. B: neurones were transfected at 10 DIV with the plasmids indicated on the left, fixed at 15 DIV, and stained with anti-CHMP2B antibody and Alexa488-labelled secondary antibody. Confocal images were acquired in both the Alexa488 and mCherry channels. Representative images of transfected neurones are displayed. Control: empty pSuper-mCherry vector. Note the drop in immunofluorescence in the shRNA-expressing neuron (arrow), compared to surrounding cells; and the decreased green vs. red ratio (overlay), compared to control or rescued cells. Scale bar: 10 μm. C: maximal intensity projections of image stacks, showing representative dendritic segments visualized by mCherry fluorescence in neurones transfected with the indicated plasmids.
    Transfection Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1822 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore transfection medium
    Influence of MALAT1 and HIF-2α on the neoplastic capacity of transformed L-02 cells Abbreviations : C-L-02 , passage control L-02 cells; T - L-02 , arsenite-transformed L-02 cells. T-L-02 cells were infected with a non-targeting control vector (sh-NC) or MALAT1 shRNA (sh-MALAT1), inducing puromycin resistance. Cells were cultured for at least 2 weeks in the presence of puromycin (5 μg/mL) before the following experiments. ( A ) T-L-02/shNC cells and T-L-02/sh-MALAT1 cells. Fluorescent microscopy. Sh-MALAT1 and sh-NC were transfected into T-L-02 cells. At 24 h after <t>transfection,</t> fluorescent microscopy showed emission green fluorescence. ( B ) qRT-PCR validated the downregulation of MALAT1 after shRNA knockdown in T-L-02 cells, * P
    Transfection Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 877 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    transfection medium - by Bioz Stars, 2020-08
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    99
    Thermo Fisher optimem transfection medium
    Influence of MALAT1 and HIF-2α on the neoplastic capacity of transformed L-02 cells Abbreviations : C-L-02 , passage control L-02 cells; T - L-02 , arsenite-transformed L-02 cells. T-L-02 cells were infected with a non-targeting control vector (sh-NC) or MALAT1 shRNA (sh-MALAT1), inducing puromycin resistance. Cells were cultured for at least 2 weeks in the presence of puromycin (5 μg/mL) before the following experiments. ( A ) T-L-02/shNC cells and T-L-02/sh-MALAT1 cells. Fluorescent microscopy. Sh-MALAT1 and sh-NC were transfected into T-L-02 cells. At 24 h after <t>transfection,</t> fluorescent microscopy showed emission green fluorescence. ( B ) qRT-PCR validated the downregulation of MALAT1 after shRNA knockdown in T-L-02 cells, * P
    Optimem Transfection Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    R&D Systems sirna transfection
    Human recombinant VEGF 165 promoted HNSCC cell proliferation following suppression of endogenous VEGF production. A and B : HNSCC cells were successfully transfected with NeoFX /FAM-labeled <t>siRNA</t> (72 h after <t>transfection,</t> 200X). C : Serum deprived HNSCC cells
    Sirna Transfection, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Roche opti mem plus x tremegene shrna transfection reagent
    Human recombinant VEGF 165 promoted HNSCC cell proliferation following suppression of endogenous VEGF production. A and B : HNSCC cells were successfully transfected with NeoFX /FAM-labeled <t>siRNA</t> (72 h after <t>transfection,</t> 200X). C : Serum deprived HNSCC cells
    Opti Mem Plus X Tremegene Shrna Transfection Reagent, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Slug regulation-induced processing of Slug and PUMA. ( A ) The protein fractions of Slug in QBC939, RBE, ICC-9810, and FRH 0201 cells were subjected to Western blot analysis. FRH 0201 exhibited the lowest expression level of Slug and QBC939 exhibited the highest expression level of Slug. ( B ) The protein fractions of Slug, E-cadherin, and PUMA in QBC939 cells after transfection with Slug siRNA for 48 h were subjected to Western blot analysis. Slug expression was barely detectable compared with parental cells (** P

    Journal: International Journal of Molecular Sciences

    Article Title: RNA Interference Targeting Slug Increases Cholangiocarcinoma Cell Sensitivity to Cisplatin via Upregulating PUMA

    doi: 10.3390/ijms12010385

    Figure Lengend Snippet: Slug regulation-induced processing of Slug and PUMA. ( A ) The protein fractions of Slug in QBC939, RBE, ICC-9810, and FRH 0201 cells were subjected to Western blot analysis. FRH 0201 exhibited the lowest expression level of Slug and QBC939 exhibited the highest expression level of Slug. ( B ) The protein fractions of Slug, E-cadherin, and PUMA in QBC939 cells after transfection with Slug siRNA for 48 h were subjected to Western blot analysis. Slug expression was barely detectable compared with parental cells (** P

    Article Snippet: For each transfection, we added 0.8 mL siRNA transfection medium to each tube containing the siRNA transfection reagent mixture (Solution A + Solution B) and then mixed gently and overlayed the mixture onto the washed cells.

    Techniques: Immunocytochemistry, Western Blot, Expressing, Transfection

    Effects of siRNA BGR1 knockdown on the ATP depletion–induced expression of TNF- α and MCP-1 genes in proximal tubule HK-2 cell culture. After transfection with either BRG1 siRNA (■) or noncomplementary (NC) siRNA ( ), HK-2 cells

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: BRG1 Increases Transcription of Proinflammatory Genes in Renal Ischemia

    doi: 10.1681/ASN.2009010118

    Figure Lengend Snippet: Effects of siRNA BGR1 knockdown on the ATP depletion–induced expression of TNF- α and MCP-1 genes in proximal tubule HK-2 cell culture. After transfection with either BRG1 siRNA (■) or noncomplementary (NC) siRNA ( ), HK-2 cells

    Article Snippet: Lipofectamine 2000 (Invitrogen, Carlsbad, CA) and either BRG1 siRNA (sc-29827; Santa Cruz Biotechnology, Santa Cruz, CA) or NC siRNA (sc-37007; Santa Cruz Biotechnology) Transfection Reagents was diluted separately with siRNA Transfection Medium (Opti-MEM I; Invitrogen).

    Techniques: Expressing, Cell Culture, Transfection

    Effects of siRNA BGR1 knockdown on the ATP depletion–induced co-recruitment of Pol II and BRG1 to the MCP-1 gene in HK-2 cells. Cells were transfected with either BRG1 siRNA (■) specific or NC siRNA ( ). After transfection, HK-2 cells were

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: BRG1 Increases Transcription of Proinflammatory Genes in Renal Ischemia

    doi: 10.1681/ASN.2009010118

    Figure Lengend Snippet: Effects of siRNA BGR1 knockdown on the ATP depletion–induced co-recruitment of Pol II and BRG1 to the MCP-1 gene in HK-2 cells. Cells were transfected with either BRG1 siRNA (■) specific or NC siRNA ( ). After transfection, HK-2 cells were

    Article Snippet: Lipofectamine 2000 (Invitrogen, Carlsbad, CA) and either BRG1 siRNA (sc-29827; Santa Cruz Biotechnology, Santa Cruz, CA) or NC siRNA (sc-37007; Santa Cruz Biotechnology) Transfection Reagents was diluted separately with siRNA Transfection Medium (Opti-MEM I; Invitrogen).

    Techniques: Transfection

    Effects of siRNA-induced BGR1 knockdown on the ATP depletion–induced co-recruitment of Pol II and BRG1 to the TNF- α gene in HK-2 cells. Cells were transfected with either BRG1 siRNA (■) specific or NC siRNA ( ). After transfection,

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: BRG1 Increases Transcription of Proinflammatory Genes in Renal Ischemia

    doi: 10.1681/ASN.2009010118

    Figure Lengend Snippet: Effects of siRNA-induced BGR1 knockdown on the ATP depletion–induced co-recruitment of Pol II and BRG1 to the TNF- α gene in HK-2 cells. Cells were transfected with either BRG1 siRNA (■) specific or NC siRNA ( ). After transfection,

    Article Snippet: Lipofectamine 2000 (Invitrogen, Carlsbad, CA) and either BRG1 siRNA (sc-29827; Santa Cruz Biotechnology, Santa Cruz, CA) or NC siRNA (sc-37007; Santa Cruz Biotechnology) Transfection Reagents was diluted separately with siRNA Transfection Medium (Opti-MEM I; Invitrogen).

    Techniques: Transfection

    CD74 knockdown by CD74 siRNA transfection reagent in CAMA-1 and MDA-MB-231 cells; A) and B) siRNA-mediated knockdown of CD74 expression in CAMA-1 and MDA-MB-231 cells were detected by Western blot; An approximately two to five (siRNA) fold weaker signal of CD74 protein expression is apparent, as compared to the negative control siRNA group normalized to the expression of α-Tubulin; C) and D) Confocal images of CAMA-1 and MDA-MB-231 cells transfected with CD74 siRNA, untreated control and negative control siRNA; Data represent three different experiments

    Journal: Open Access Macedonian Journal of Medical Sciences

    Article Title: Knockdown of CD-74 in the Proliferative and Apoptotic Activity of Breast Cancer Cells

    doi: 10.3889/oamjms.2019.354

    Figure Lengend Snippet: CD74 knockdown by CD74 siRNA transfection reagent in CAMA-1 and MDA-MB-231 cells; A) and B) siRNA-mediated knockdown of CD74 expression in CAMA-1 and MDA-MB-231 cells were detected by Western blot; An approximately two to five (siRNA) fold weaker signal of CD74 protein expression is apparent, as compared to the negative control siRNA group normalized to the expression of α-Tubulin; C) and D) Confocal images of CAMA-1 and MDA-MB-231 cells transfected with CD74 siRNA, untreated control and negative control siRNA; Data represent three different experiments

    Article Snippet: For each transfection, 4 µl of CD74 siRNA duplex at dose of 80 pmols (sc-35023) (Santa Cruz Biotechnology, USA) was diluted and 4 µl of siRNA transfection reagent was added at dose of 80 pmols (sc-29528) (Santa Cruz Biotechnology, USA) into 100 µl of siRNA transfection medium (sc-36868) (Santa Cruz Biotechnology, USA) separately without serum or antibiotics.

    Techniques: Transfection, Multiple Displacement Amplification, Expressing, Western Blot, Negative Control

    CD5L serves a cytoprotective function by modulating autophagy. (A-E) To determine the role of autophagy in the anti-apoptotic actions of CD5L, hepatocytes were transfected with siRNA against ATG7 or siRNA-NT. (A) The siRNA-mediated transfection efficiency was demonstrated by reverse transcription-quantitative PCR. Each column represents the mean ± SD of three independent experiments. * P

    Journal: Experimental and Therapeutic Medicine

    Article Title: CD5L-induced activation of autophagy is associated with hepatoprotection in ischemic reperfusion injury via the CD36/ATG7 axis

    doi: 10.3892/etm.2020.8497

    Figure Lengend Snippet: CD5L serves a cytoprotective function by modulating autophagy. (A-E) To determine the role of autophagy in the anti-apoptotic actions of CD5L, hepatocytes were transfected with siRNA against ATG7 or siRNA-NT. (A) The siRNA-mediated transfection efficiency was demonstrated by reverse transcription-quantitative PCR. Each column represents the mean ± SD of three independent experiments. * P

    Article Snippet: Small interfering (si)RNA transfection Hepatocytes were resuspended (1x106 cells/ml) in serum- and antibiotic-free siRNA Transfection Medium (cat. no. sc-36868; Santa Cruz Biotechnology, Inc.).

    Techniques: Transfection, Real-time Polymerase Chain Reaction

    Inhibition of hepatocyte apoptosis by CD5L occurs via the CD36 receptor. (A-E) Hepatocytes were transfected with siRNA targeting CD36 or with siRNA-NT as a control. (A) Transfection efficiency was determined by reverse transcription-quantitative PCR. Each column represents the mean ± SD of three independent experiments. * P

    Journal: Experimental and Therapeutic Medicine

    Article Title: CD5L-induced activation of autophagy is associated with hepatoprotection in ischemic reperfusion injury via the CD36/ATG7 axis

    doi: 10.3892/etm.2020.8497

    Figure Lengend Snippet: Inhibition of hepatocyte apoptosis by CD5L occurs via the CD36 receptor. (A-E) Hepatocytes were transfected with siRNA targeting CD36 or with siRNA-NT as a control. (A) Transfection efficiency was determined by reverse transcription-quantitative PCR. Each column represents the mean ± SD of three independent experiments. * P

    Article Snippet: Small interfering (si)RNA transfection Hepatocytes were resuspended (1x106 cells/ml) in serum- and antibiotic-free siRNA Transfection Medium (cat. no. sc-36868; Santa Cruz Biotechnology, Inc.).

    Techniques: Inhibition, Transfection, Real-time Polymerase Chain Reaction

    Tenascin C contributes to NRP-1 associated migration. a. Dual immunofluorescence staining (40× magnification scale bar 10 μm) of NRP-1 and TNC on BT-474 and BT-474 NRP-1 cells indicates their colocalization in the cytoplasm. Treatment of BT-474 NRP-1 cells with TNC targeted siRNA molecules, b , reduced TNC gene expression, c , reduced migratory capacity and d , downregulated NRP-1 and vimentin expression. (TNC protein was not detected on western blot due to the lack of specific antibody for this application.) The gene expression fold change was measured by comparing the basal levels detected in the empty plasmid transfected BT-474 or in the case of the siRNA experiment, to the control siRNA treated BT-474 and normalized to β-Actin and GUSB reference gene expression. Wound healing assay images (panel d, 5× magnification, scale bar 500 μm) taken on day 0 and day 2 after siRNA transfection. Graphs represent the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value

    Journal: BMC Cancer

    Article Title: Neuropilin-1 promotes the oncogenic Tenascin-C/integrin β3 pathway and modulates chemoresistance in breast cancer cells

    doi: 10.1186/s12885-018-4446-y

    Figure Lengend Snippet: Tenascin C contributes to NRP-1 associated migration. a. Dual immunofluorescence staining (40× magnification scale bar 10 μm) of NRP-1 and TNC on BT-474 and BT-474 NRP-1 cells indicates their colocalization in the cytoplasm. Treatment of BT-474 NRP-1 cells with TNC targeted siRNA molecules, b , reduced TNC gene expression, c , reduced migratory capacity and d , downregulated NRP-1 and vimentin expression. (TNC protein was not detected on western blot due to the lack of specific antibody for this application.) The gene expression fold change was measured by comparing the basal levels detected in the empty plasmid transfected BT-474 or in the case of the siRNA experiment, to the control siRNA treated BT-474 and normalized to β-Actin and GUSB reference gene expression. Wound healing assay images (panel d, 5× magnification, scale bar 500 μm) taken on day 0 and day 2 after siRNA transfection. Graphs represent the mean ± SEM of three independent experiments. Statistical analysis using independent samples t-test, p-value

    Article Snippet: Cells were treated with 80 pmol of a pool of three human TNC-targeted siRNA (Santa Cruz) or control siRNA in transfection reagent and serum-free siRNA transfection medium (Santa Cruz) according to the manufacturer’s instructions.

    Techniques: Migration, Immunofluorescence, Staining, Expressing, Western Blot, Plasmid Preparation, Transfection, Wound Healing Assay

    Kinases Lyn and Syk selectively regulate Ras isoform’s activation through Ras-GEFs. a CD40 activates the upstream kinases Lyn and Syk in a dose-dependent manner, with lower doses activating Syk and higher doses activating Lyn. b Immunoblot analysis of total Ras and activated Ras isoforms in the lysates of un-transfected or Syk- or Lyn-specific siRNA transfected, anti-CD40 antibody (3 μg/ml) treated P388D1 cells. c Immunoblot analyses of phospho-tyrosine (p-tyr-Vav)), and Vav translocated Sos-1/2 (Tr-Sos-1/2), translocated Ras-GRP (Tr-RasGRP), CD71, syk, lyn and β-actin in the lysates of untreated or Syk siRNA or Lyn siRNA or anti-CD40 antibody (3 μg/ml) treated P388D1 cells. d Immunoblot analysis of phospho-tyrosine (p-tyr Vav) and Vav translocated Sos-1/2, translocated Ras GRP, CD71, Lyn and phospho-Lyn, Syk and phospho-Syk (p-syk) in the lysates of untreated or Syk inhibitor (Syk Inh, 3 μM; Calbiochem, San Diego, CA) or PP-1 (340 nM; BIOMOL International, PA) treated and/or anti-CD40 antibody (3 μg/ml) treated macrophages. e Analysis of the association of Syk or Lyn with Sos-1/2, Vav, and Ras-GRP in the co-immunoprecipitates from the lysates of anti-CD40 antibody (1 μg/ml, 3 μg/ml and 6 μg/ml) treated macrophages.

    Journal: Cell Communication and Signaling : CCS

    Article Title: Ras isoforms: signaling specificities in CD40 pathway

    doi: 10.1186/s12964-019-0497-1

    Figure Lengend Snippet: Kinases Lyn and Syk selectively regulate Ras isoform’s activation through Ras-GEFs. a CD40 activates the upstream kinases Lyn and Syk in a dose-dependent manner, with lower doses activating Syk and higher doses activating Lyn. b Immunoblot analysis of total Ras and activated Ras isoforms in the lysates of un-transfected or Syk- or Lyn-specific siRNA transfected, anti-CD40 antibody (3 μg/ml) treated P388D1 cells. c Immunoblot analyses of phospho-tyrosine (p-tyr-Vav)), and Vav translocated Sos-1/2 (Tr-Sos-1/2), translocated Ras-GRP (Tr-RasGRP), CD71, syk, lyn and β-actin in the lysates of untreated or Syk siRNA or Lyn siRNA or anti-CD40 antibody (3 μg/ml) treated P388D1 cells. d Immunoblot analysis of phospho-tyrosine (p-tyr Vav) and Vav translocated Sos-1/2, translocated Ras GRP, CD71, Lyn and phospho-Lyn, Syk and phospho-Syk (p-syk) in the lysates of untreated or Syk inhibitor (Syk Inh, 3 μM; Calbiochem, San Diego, CA) or PP-1 (340 nM; BIOMOL International, PA) treated and/or anti-CD40 antibody (3 μg/ml) treated macrophages. e Analysis of the association of Syk or Lyn with Sos-1/2, Vav, and Ras-GRP in the co-immunoprecipitates from the lysates of anti-CD40 antibody (1 μg/ml, 3 μg/ml and 6 μg/ml) treated macrophages.

    Article Snippet: Control siRNA, H-Ras siRNA, N-Ras siRNA, K-Ras siRNA, Ras-GRP siRNA, Ras-GRF siRNA, Sos1/2 siRNA, Vav siRNA, Lyn siRNA, Syk siRNA transfection medium and transfection reagents were purchased from Santa Cruz biotechnology.

    Techniques: Activation Assay, Transfection

    Unsupervised hierarchical clustering of miRNAs reveals relationship of groups. Experimental groups were largely recapitulated by unsupervised hierarchical clustering of all results from OpenArray miRNA profiling. Clustering was performed by Pearson correlation

    Journal: Cancer Biology & Therapy

    Article Title: Dietary flaxseed modulates the miRNA profile in irradiated and non-irradiated murine lungs

    doi: 10.4161/cbt.28905

    Figure Lengend Snippet: Unsupervised hierarchical clustering of miRNAs reveals relationship of groups. Experimental groups were largely recapitulated by unsupervised hierarchical clustering of all results from OpenArray miRNA profiling. Clustering was performed by Pearson correlation

    Article Snippet: miRNA profiling was conducted using the medium-throughput rodent miRNA “OpenArray” platform from Life Technologies.

    Techniques:

    knock-down of endogenous CHMP2B in hippocampal neurones. A: to verify the efficacy of the shRNA-encoding plasmid, the indicated plasmids were transfected in rodent (BHK) cells and equal protein amounts of transfected cell lysates were analysed by Western immunoblotting with anti-CHMP2B antibody, or with anti-actin as a control. Lane 1: empty pSuper-mCherry plasmid; lane 2: shRNA-expressing plasmid; lane 3: shRNA-expressing plasmid cotransfected with vector encoding CHMP2B* (native CHMP2B cDNA with silent mutations at the siRNA target site). Note that the CHMP2B protein remaining after transfection of the shRNA plasmid largely originates from non-transfected cells in the culture. B: neurones were transfected at 10 DIV with the plasmids indicated on the left, fixed at 15 DIV, and stained with anti-CHMP2B antibody and Alexa488-labelled secondary antibody. Confocal images were acquired in both the Alexa488 and mCherry channels. Representative images of transfected neurones are displayed. Control: empty pSuper-mCherry vector. Note the drop in immunofluorescence in the shRNA-expressing neuron (arrow), compared to surrounding cells; and the decreased green vs. red ratio (overlay), compared to control or rescued cells. Scale bar: 10 μm. C: maximal intensity projections of image stacks, showing representative dendritic segments visualized by mCherry fluorescence in neurones transfected with the indicated plasmids.

    Journal: Journal of Cell Science

    Article Title: CHMP2B mutants linked to frontotemporal dementia impair maturation of dendritic spines

    doi: 10.1242/jcs.068817

    Figure Lengend Snippet: knock-down of endogenous CHMP2B in hippocampal neurones. A: to verify the efficacy of the shRNA-encoding plasmid, the indicated plasmids were transfected in rodent (BHK) cells and equal protein amounts of transfected cell lysates were analysed by Western immunoblotting with anti-CHMP2B antibody, or with anti-actin as a control. Lane 1: empty pSuper-mCherry plasmid; lane 2: shRNA-expressing plasmid; lane 3: shRNA-expressing plasmid cotransfected with vector encoding CHMP2B* (native CHMP2B cDNA with silent mutations at the siRNA target site). Note that the CHMP2B protein remaining after transfection of the shRNA plasmid largely originates from non-transfected cells in the culture. B: neurones were transfected at 10 DIV with the plasmids indicated on the left, fixed at 15 DIV, and stained with anti-CHMP2B antibody and Alexa488-labelled secondary antibody. Confocal images were acquired in both the Alexa488 and mCherry channels. Representative images of transfected neurones are displayed. Control: empty pSuper-mCherry vector. Note the drop in immunofluorescence in the shRNA-expressing neuron (arrow), compared to surrounding cells; and the decreased green vs. red ratio (overlay), compared to control or rescued cells. Scale bar: 10 μm. C: maximal intensity projections of image stacks, showing representative dendritic segments visualized by mCherry fluorescence in neurones transfected with the indicated plasmids.

    Article Snippet: 0.5 μg CHMP2B plasmid (or control vector) was mixed with 2.5 μg mCherry, GFP, or LC3-GFP plasmid, diluted into 250 μl transfection medium, and combined with 1.5 μl of “Plus” Reagent and then 4 μl Lipofectamine LTX (Invitrogen).

    Techniques: shRNA, Plasmid Preparation, Transfection, Western Blot, Expressing, Staining, Immunofluorescence, Fluorescence

    electrophysiological effects of wild-type and mutant CHMP2B. Miniature excitatory currents were recorded from transfected neurones under voltage clamp in the whole-cell configuration, in the presence of tetrodotoxin and D-APV to isolate AMPA receptor-mediated spontaneous synaptic currents (control: n=4 cells; CHMP2B wt : n=6 cells; CHMP2B Δ10 : n=6 cells). A: representative sample traces from neurones co-transfected with GFP and control plasmid (top), CHMP2B wt (middle), and CHMP2B Δ10 (bottom). B: mean duration of inter-event intervals along sequences of mEPSCs pooled from equivalently transfected neurones (control: n=516 events; CHMP2B wt : n=768 events; CHMP2B Δ10 : n=768 events). Compared to control, intervals were significantly longer in mEPSCs from CHMP2B Δ10 -expressing neurones (*p=0.00001), but not from CHMP2B wt neurones (p=0.05, alpha=0.017). C: mean amplitude of mEPSCs from the different transfection groups. The overall variation of mean amplitude as a function of transfected plasmid was significant (ANOVA: F 2,2049 = 3.04, p=0.048); the specific difference between the means of CHMP2B Δ10 and control was barely below threshold (*p=0.026 with Bonferroni-corrected alpha=0.025). D: cumulative histogram of mEPSC amplitudes; same dataset as in C. * indicates significant reduction in the top quintile in the case of CHMP2B Δ10 (see text and panel E). F: histogram of mEPSC amplitudes showing the specific reduction in the proportion of large currents ( > 20 pA) and the increase in small currents. S, i

    Journal: Journal of Cell Science

    Article Title: CHMP2B mutants linked to frontotemporal dementia impair maturation of dendritic spines

    doi: 10.1242/jcs.068817

    Figure Lengend Snippet: electrophysiological effects of wild-type and mutant CHMP2B. Miniature excitatory currents were recorded from transfected neurones under voltage clamp in the whole-cell configuration, in the presence of tetrodotoxin and D-APV to isolate AMPA receptor-mediated spontaneous synaptic currents (control: n=4 cells; CHMP2B wt : n=6 cells; CHMP2B Δ10 : n=6 cells). A: representative sample traces from neurones co-transfected with GFP and control plasmid (top), CHMP2B wt (middle), and CHMP2B Δ10 (bottom). B: mean duration of inter-event intervals along sequences of mEPSCs pooled from equivalently transfected neurones (control: n=516 events; CHMP2B wt : n=768 events; CHMP2B Δ10 : n=768 events). Compared to control, intervals were significantly longer in mEPSCs from CHMP2B Δ10 -expressing neurones (*p=0.00001), but not from CHMP2B wt neurones (p=0.05, alpha=0.017). C: mean amplitude of mEPSCs from the different transfection groups. The overall variation of mean amplitude as a function of transfected plasmid was significant (ANOVA: F 2,2049 = 3.04, p=0.048); the specific difference between the means of CHMP2B Δ10 and control was barely below threshold (*p=0.026 with Bonferroni-corrected alpha=0.025). D: cumulative histogram of mEPSC amplitudes; same dataset as in C. * indicates significant reduction in the top quintile in the case of CHMP2B Δ10 (see text and panel E). F: histogram of mEPSC amplitudes showing the specific reduction in the proportion of large currents ( > 20 pA) and the increase in small currents. S, i

    Article Snippet: 0.5 μg CHMP2B plasmid (or control vector) was mixed with 2.5 μg mCherry, GFP, or LC3-GFP plasmid, diluted into 250 μl transfection medium, and combined with 1.5 μl of “Plus” Reagent and then 4 μl Lipofectamine LTX (Invitrogen).

    Techniques: Mutagenesis, Transfection, Plasmid Preparation, Expressing

    Influence of MALAT1 and HIF-2α on the neoplastic capacity of transformed L-02 cells Abbreviations : C-L-02 , passage control L-02 cells; T - L-02 , arsenite-transformed L-02 cells. T-L-02 cells were infected with a non-targeting control vector (sh-NC) or MALAT1 shRNA (sh-MALAT1), inducing puromycin resistance. Cells were cultured for at least 2 weeks in the presence of puromycin (5 μg/mL) before the following experiments. ( A ) T-L-02/shNC cells and T-L-02/sh-MALAT1 cells. Fluorescent microscopy. Sh-MALAT1 and sh-NC were transfected into T-L-02 cells. At 24 h after transfection, fluorescent microscopy showed emission green fluorescence. ( B ) qRT-PCR validated the downregulation of MALAT1 after shRNA knockdown in T-L-02 cells, * P

    Journal: Oncotarget

    Article Title: A MALAT1/HIF-2α feedback loop contributes to arsenite carcinogenesis

    doi: 10.18632/oncotarget.6806

    Figure Lengend Snippet: Influence of MALAT1 and HIF-2α on the neoplastic capacity of transformed L-02 cells Abbreviations : C-L-02 , passage control L-02 cells; T - L-02 , arsenite-transformed L-02 cells. T-L-02 cells were infected with a non-targeting control vector (sh-NC) or MALAT1 shRNA (sh-MALAT1), inducing puromycin resistance. Cells were cultured for at least 2 weeks in the presence of puromycin (5 μg/mL) before the following experiments. ( A ) T-L-02/shNC cells and T-L-02/sh-MALAT1 cells. Fluorescent microscopy. Sh-MALAT1 and sh-NC were transfected into T-L-02 cells. At 24 h after transfection, fluorescent microscopy showed emission green fluorescence. ( B ) qRT-PCR validated the downregulation of MALAT1 after shRNA knockdown in T-L-02 cells, * P

    Article Snippet: After 24 h of transfection, the transfection medium was replaced with medium containing puromycin (5 μg/ml; Sigma-Aldrich) for at least 2 weeks before usage to select stable cell pools.

    Techniques: Transformation Assay, Infection, Plasmid Preparation, shRNA, Cell Culture, Microscopy, Transfection, Fluorescence, Quantitative RT-PCR

    Human recombinant VEGF 165 promoted HNSCC cell proliferation following suppression of endogenous VEGF production. A and B : HNSCC cells were successfully transfected with NeoFX /FAM-labeled siRNA (72 h after transfection, 200X). C : Serum deprived HNSCC cells

    Journal:

    Article Title: Human head and neck squamous cell carcinoma cells are both targets and effectors for the angiogenic cytokine, VEGF

    doi: 10.1002/jcb.21920

    Figure Lengend Snippet: Human recombinant VEGF 165 promoted HNSCC cell proliferation following suppression of endogenous VEGF production. A and B : HNSCC cells were successfully transfected with NeoFX /FAM-labeled siRNA (72 h after transfection, 200X). C : Serum deprived HNSCC cells

    Article Snippet: Samples of the conditioned medium obtained at 24, 48, 72, and 96 h after siRNA transfection were analyzed by ELISA (Human VEGF DuoSet ELISA Development Kit, R & D Systems, Minneapolis, MN) to determine the concentration of VEGF protein secreted by HUVEC and HNSCC cells.

    Techniques: Recombinant, Transfection, Labeling