bcl 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna
    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the <t>Bcl-2</t> gene.
    Bcl 2 Sirna, supplied by Cell Signaling Technology Inc, 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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    Images

    1) Product Images from "An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma"

    Article Title: An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S32900

    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.
    Figure Legend Snippet: T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.

    Techniques Used: Imaging, Modification, Magnetic Resonance Imaging, Reverse Transcription Polymerase Chain Reaction

    Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.
    Figure Legend Snippet: Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription Polymerase Chain Reaction

    Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.
    Figure Legend Snippet: Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.

    Techniques Used:

    bcl 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna
    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the <t>Bcl-2</t> gene.
    Bcl 2 Sirna, supplied by Cell Signaling Technology Inc, 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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    Images

    1) Product Images from "An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma"

    Article Title: An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S32900

    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.
    Figure Legend Snippet: T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.

    Techniques Used: Imaging, Modification, Magnetic Resonance Imaging, Reverse Transcription Polymerase Chain Reaction

    Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.
    Figure Legend Snippet: Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription Polymerase Chain Reaction

    Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.
    Figure Legend Snippet: Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.

    Techniques Used:

    bcl 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna
    Bcl 2 Sirna, supplied by Cell Signaling Technology Inc, 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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    signalsilence ampkα2 sirna ii  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc signalsilence ampkα2 sirna ii
    Signalsilence Ampkα2 Sirna Ii, supplied by Cell Signaling Technology Inc, 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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    6630s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc 6630s
    KEY RESOURCES TABLE
    6630s, supplied by Cell Signaling Technology Inc, 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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    1) Product Images from "The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress"

    Article Title: The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress

    Journal: Cell metabolism

    doi: 10.1016/j.cmet.2018.06.008

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, SYBR Green Assay, Reporter Assay, Over Expression, Sequencing, Luciferase, Software, Real-time Polymerase Chain Reaction

    bcl 2 sirna kit  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna kit
    Bcl 2 Sirna Kit, supplied by Cell Signaling Technology Inc, 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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    pka c α sirna ii  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pka c α sirna ii
    Pka C α Sirna Ii, supplied by Cell Signaling Technology Inc, 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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    tp53 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc tp53 2 sirna
    Flow chart of the comprehensive screening for synthetic lethal genes interacting with <t>p53</t> R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.
    Tp53 2 Sirna, supplied by Cell Signaling Technology Inc, 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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    1) Product Images from "High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H"

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    Journal: Oncology Reports

    doi: 10.3892/or.2013.2953

    Flow chart of the comprehensive screening for synthetic lethal genes interacting with p53 R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.
    Figure Legend Snippet: Flow chart of the comprehensive screening for synthetic lethal genes interacting with p53 R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.

    Techniques Used: shRNA, Hybridization

    Suppression of candidate genes in PC3 cells with p53 R175H expression. Ninety-six hours after transfection of plasmid pCR259 or pCR259-R175H in PC3 cells, (A) p53 R175H expression was confirmed by western blotting, and (B) cell numbers were measured by performing cell proliferation assays. The vertical axis corresponds to the absorbance ratio of (candidate gene siRNA transfected cells)/(negative control siRNA transfected cells) for 5 candidate genes. Values shown are means ± SD (n=3). * p<0.05 between p53 R175H null and p53 R175H expression. (C) Western blot analysis of Id1 and p53 R175H. Id1 expression was unchanged by R175H expression.
    Figure Legend Snippet: Suppression of candidate genes in PC3 cells with p53 R175H expression. Ninety-six hours after transfection of plasmid pCR259 or pCR259-R175H in PC3 cells, (A) p53 R175H expression was confirmed by western blotting, and (B) cell numbers were measured by performing cell proliferation assays. The vertical axis corresponds to the absorbance ratio of (candidate gene siRNA transfected cells)/(negative control siRNA transfected cells) for 5 candidate genes. Values shown are means ± SD (n=3). * p<0.05 between p53 R175H null and p53 R175H expression. (C) Western blot analysis of Id1 and p53 R175H. Id1 expression was unchanged by R175H expression.

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Western Blot, Negative Control

    Comparison of cell growth inhibition by ID1 suppression alone and ID1 / TP53 double suppression in cells expressing p53 R175H, wild-type p53, and TP53 -null cells. (A) ID1 and TP53 siRNA were co-transfected into SKBr3 cells to make a final concentration of 100 nM, and cell numbers were measured by performing cell proliferation assays on days 2 and 4. The vertical axis corresponds to the absorbance of the cell proliferation assay. Top of each figure represents expression inhibition of Id1 or p53 by each siRNA. Furthermore, other siRNAs of ID1 ( ID1 - 2 ) (B) and TP53 ( TP53 -2) (C) targeting different sites were transfected into SKBr3 cells. The same experiments were conducted in LS123 and HCC1395 cells, p53 R175H expressing cell lines (D and E), HCT116 cells expressing wild-type p53 (F), and the TP53 -null cell line PC3 (G). Values shown are means ± SD (n=3) in (A and E). * p<0.05, ** p<0.01, between ID1 suppression alone and ID1 / TP53 double suppression. (H) Western blot analysis of Id1 and p53 R175H. Knockdown level of Id1 was not rescued by ID1 / TP53 double suppression in SKBr3 cells.
    Figure Legend Snippet: Comparison of cell growth inhibition by ID1 suppression alone and ID1 / TP53 double suppression in cells expressing p53 R175H, wild-type p53, and TP53 -null cells. (A) ID1 and TP53 siRNA were co-transfected into SKBr3 cells to make a final concentration of 100 nM, and cell numbers were measured by performing cell proliferation assays on days 2 and 4. The vertical axis corresponds to the absorbance of the cell proliferation assay. Top of each figure represents expression inhibition of Id1 or p53 by each siRNA. Furthermore, other siRNAs of ID1 ( ID1 - 2 ) (B) and TP53 ( TP53 -2) (C) targeting different sites were transfected into SKBr3 cells. The same experiments were conducted in LS123 and HCC1395 cells, p53 R175H expressing cell lines (D and E), HCT116 cells expressing wild-type p53 (F), and the TP53 -null cell line PC3 (G). Values shown are means ± SD (n=3) in (A and E). * p<0.05, ** p<0.01, between ID1 suppression alone and ID1 / TP53 double suppression. (H) Western blot analysis of Id1 and p53 R175H. Knockdown level of Id1 was not rescued by ID1 / TP53 double suppression in SKBr3 cells.

    Techniques Used: Inhibition, Expressing, Transfection, Concentration Assay, Proliferation Assay, Western Blot

    ID1 suppression in PC3 cells expressing p53 R273H and ID1 / TP53 double suppression in cells expressing p53 R273H. (A) The plasmid pCR259-R273H was transfected into PC3 cells for 96 h, and total cell numbers were measured by performing cell proliferation assays. ID1 siRNA and TP53 siRNA were co-transfected into HT-29 cells expressing endogenous p53 R273H (B) and SW480 expressing endogenous p53 R273H/R309S double mutant (C). Cell counts were measured by performing cell proliferation assays on days 2 and 4. The vertical axis is same as in . The top right of each figure is the same as that in . Values shown are means ± SD (n=3).
    Figure Legend Snippet: ID1 suppression in PC3 cells expressing p53 R273H and ID1 / TP53 double suppression in cells expressing p53 R273H. (A) The plasmid pCR259-R273H was transfected into PC3 cells for 96 h, and total cell numbers were measured by performing cell proliferation assays. ID1 siRNA and TP53 siRNA were co-transfected into HT-29 cells expressing endogenous p53 R273H (B) and SW480 expressing endogenous p53 R273H/R309S double mutant (C). Cell counts were measured by performing cell proliferation assays on days 2 and 4. The vertical axis is same as in . The top right of each figure is the same as that in . Values shown are means ± SD (n=3).

    Techniques Used: Expressing, Plasmid Preparation, Transfection, Mutagenesis

    FACS analysis of SKBr3 (A), HCT116 (B) and PC3 (C) cells with ID1 and TP53 double suppression. FACS analysis with or without ID1 siRNA (100 nM) and TP53 siRNA (100 nM). Values of the sub-G1 fraction are a portion of the total population, and values of G1, S, and G2/M fractions are portions of the population excluding the sub-G1 fraction. Values shown are means ± SD (n=6). * p<0.05, ** p<0.01, between negative control and ID1 suppression or between ID1 suppression and ID1 / TP53 double suppression.
    Figure Legend Snippet: FACS analysis of SKBr3 (A), HCT116 (B) and PC3 (C) cells with ID1 and TP53 double suppression. FACS analysis with or without ID1 siRNA (100 nM) and TP53 siRNA (100 nM). Values of the sub-G1 fraction are a portion of the total population, and values of G1, S, and G2/M fractions are portions of the population excluding the sub-G1 fraction. Values shown are means ± SD (n=6). * p<0.05, ** p<0.01, between negative control and ID1 suppression or between ID1 suppression and ID1 / TP53 double suppression.

    Techniques Used: Negative Control

    bcl 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna
    Cytotoxic activity of S1 against a panel of SCLC cell lines associated with <t>Bcl-2</t> expression and phosphorylation. (A) Cell viability EC50 values of various SCLC cell lines following treatment for 48 h with S1 or ABT-737 were determined by MTS assay. Columns, average (n = 3) of triplicate experiments; bars, SD. (B) Western blot analysis of the Bcl-2 family proteins indicated in the 11 cell lines. Cell lines are arranged according to their resistance to S1.
    Bcl 2 Sirna, supplied by Cell Signaling Technology Inc, 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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    1) Product Images from "Resistance to BH3 mimetic S1 in SCLC cells that up-regulate and phosphorylate Bcl-2 through ERK1/2"

    Article Title: Resistance to BH3 mimetic S1 in SCLC cells that up-regulate and phosphorylate Bcl-2 through ERK1/2

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.12243

    Cytotoxic activity of S1 against a panel of SCLC cell lines associated with Bcl-2 expression and phosphorylation. (A) Cell viability EC50 values of various SCLC cell lines following treatment for 48 h with S1 or ABT-737 were determined by MTS assay. Columns, average (n = 3) of triplicate experiments; bars, SD. (B) Western blot analysis of the Bcl-2 family proteins indicated in the 11 cell lines. Cell lines are arranged according to their resistance to S1.
    Figure Legend Snippet: Cytotoxic activity of S1 against a panel of SCLC cell lines associated with Bcl-2 expression and phosphorylation. (A) Cell viability EC50 values of various SCLC cell lines following treatment for 48 h with S1 or ABT-737 were determined by MTS assay. Columns, average (n = 3) of triplicate experiments; bars, SD. (B) Western blot analysis of the Bcl-2 family proteins indicated in the 11 cell lines. Cell lines are arranged according to their resistance to S1.

    Techniques Used: Activity Assay, Expressing, MTS Assay, Western Blot

    Bcl-2 is transcriptionally up-regulated in H1688-acquired resistant cells. (A) EC50 values (S1 and ABT-737 48 h) and expressions of Bcl-2 family proteins of parental and S1-acquired resistant H1688 cells. Columns, average (n = 3) of triplicate experiments; bars, SD. (B) Bcl-2 transcript level was analysed by quantitative PCR in H1688-derived cells. H1688-derived resistant cells were cultured in the absence of S1 for 3 weeks and cells were treated or not with 3 μg·mL−1 actinomycin D (Act-D) for 1 h before RNA was isolated. (C) Bcl-2 is up-regulated in both transcriptional, protein and phosphorylation levels in parental H1688 and resistant H1688-SR10 cells after transient treatment with S1. H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688 and H1688-SR10, 10 μM for H1688-SR10) for indicated time and Bcl-2 transcript level was analysed by quantitative PCR. H1688-SR10 cells were removed from S1-containing media for 3 weeks. H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688, 10 μM for H1688-SR10) or DMSO for indicated time. H1688 and H1688-SR10 cells were treated actinomycin D (3 μg·mL−1) for 1 h before the addition of S1. Whole-cell lysates were made after treatment and analysed by immunoblot. Columns, average (n = 3) of triplicate experiments; bars, SD. (D) The indicated cell lines were transfected with control (Ctrl) or Bcl-2 siRNA for 48 h and then treated with and without 10 μM S1 or ABT-737 for an additional 16 h. Protein levels were examined by Western blot. Cell viability was assessed by MTS assay.
    Figure Legend Snippet: Bcl-2 is transcriptionally up-regulated in H1688-acquired resistant cells. (A) EC50 values (S1 and ABT-737 48 h) and expressions of Bcl-2 family proteins of parental and S1-acquired resistant H1688 cells. Columns, average (n = 3) of triplicate experiments; bars, SD. (B) Bcl-2 transcript level was analysed by quantitative PCR in H1688-derived cells. H1688-derived resistant cells were cultured in the absence of S1 for 3 weeks and cells were treated or not with 3 μg·mL−1 actinomycin D (Act-D) for 1 h before RNA was isolated. (C) Bcl-2 is up-regulated in both transcriptional, protein and phosphorylation levels in parental H1688 and resistant H1688-SR10 cells after transient treatment with S1. H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688 and H1688-SR10, 10 μM for H1688-SR10) for indicated time and Bcl-2 transcript level was analysed by quantitative PCR. H1688-SR10 cells were removed from S1-containing media for 3 weeks. H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688, 10 μM for H1688-SR10) or DMSO for indicated time. H1688 and H1688-SR10 cells were treated actinomycin D (3 μg·mL−1) for 1 h before the addition of S1. Whole-cell lysates were made after treatment and analysed by immunoblot. Columns, average (n = 3) of triplicate experiments; bars, SD. (D) The indicated cell lines were transfected with control (Ctrl) or Bcl-2 siRNA for 48 h and then treated with and without 10 μM S1 or ABT-737 for an additional 16 h. Protein levels were examined by Western blot. Cell viability was assessed by MTS assay.

    Techniques Used: Real-time Polymerase Chain Reaction, Derivative Assay, Cell Culture, Isolation, Western Blot, Transfection, MTS Assay

    S1 induces up-regulation of Bcl-2 by an ER stress-induced ERK1/2 activation-dependent mechanism. (A) H1688-derived cells were incubated with 5 μM PD98059 for 16 h. DMSO treatment (left) was used as a control. Whole cell lysates were prepared and Western blots were probed using specific antibodies against phosphorylated ERK1/2, whole ERK1/2 and Bcl-2. Blot shows ERK phosphorylation correlated with up-regulation of Bcl-2 (left). (B) Whole-cell lysates from 11 SCLC cell lines were analysed with Western blot for the levels of phosphorylated ERK1/2 and ERK1/2. Cell lines are arranged according to their resistance to S1. (C) H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688 and 10 μM for H1688-SR10) alone or together with 5 μM PD98059 for 16 h. Protein levels were examined by Western blot. H1688-SR10 cells were cultured in the absence of S1 for 3 weeks before used. (D) H209 and H740 cells were treated with 400 nM S1 alone or together with 5 μM PD98059 for 16 h. ABT-737 (100 nM) was tested in parallel. DMSO was used as a control. Whole cell lysates were prepared and Western blots were probed using indicated antibodies. (E) The indicated cell lines were transfected with control (Ctrl) or ERK1/2 siRNA for 48 h and then treated with and without 10 μM S1 or ABT-737 for an additional 16 h. Protein levels were examined by Western blot. H1688-SR10 and H740 cells viability were assessed by MTS assay.
    Figure Legend Snippet: S1 induces up-regulation of Bcl-2 by an ER stress-induced ERK1/2 activation-dependent mechanism. (A) H1688-derived cells were incubated with 5 μM PD98059 for 16 h. DMSO treatment (left) was used as a control. Whole cell lysates were prepared and Western blots were probed using specific antibodies against phosphorylated ERK1/2, whole ERK1/2 and Bcl-2. Blot shows ERK phosphorylation correlated with up-regulation of Bcl-2 (left). (B) Whole-cell lysates from 11 SCLC cell lines were analysed with Western blot for the levels of phosphorylated ERK1/2 and ERK1/2. Cell lines are arranged according to their resistance to S1. (C) H1688 and H1688-SR10 cells were treated with S1 (400 nM for H1688 and 10 μM for H1688-SR10) alone or together with 5 μM PD98059 for 16 h. Protein levels were examined by Western blot. H1688-SR10 cells were cultured in the absence of S1 for 3 weeks before used. (D) H209 and H740 cells were treated with 400 nM S1 alone or together with 5 μM PD98059 for 16 h. ABT-737 (100 nM) was tested in parallel. DMSO was used as a control. Whole cell lysates were prepared and Western blots were probed using indicated antibodies. (E) The indicated cell lines were transfected with control (Ctrl) or ERK1/2 siRNA for 48 h and then treated with and without 10 μM S1 or ABT-737 for an additional 16 h. Protein levels were examined by Western blot. H1688-SR10 and H740 cells viability were assessed by MTS assay.

    Techniques Used: Activation Assay, Derivative Assay, Incubation, Western Blot, Cell Culture, Transfection, MTS Assay

    Bcl-2 is phosphorylated by ERK1/2 in H1688-derived cells and contributes to the resistance to S1. (A) PD98059 was able to block Bcl-2 phosphorylation and reduce the pBcl-2/Bcl-2 ratios. H1688-derived H1688-SR6, H1688-SR10 cell, H1048 and H740 cells were incubated with 5 μM PD98059 for 16 h. DMSO was used as a control. Total protein extracts (50 μg) from these cells were analysed by Western blot for pBcl-2 and Bcl-2 expression. The mean of pBcl-2/Bcl-2 ratio is shown for each cell line. (B) pBcl-2 sequestrated more pro-apoptotic proteins in acquired resistant cells. Immunoprecipitation of H1688-derived cell lysates. Mcl-1, Bcl-2 and pBcl-2 immunoprecipitations were performed, and immunoprecipitated fractions were analysed by Western blotting for the indicated proteins. (C) H1688-SR10 cells were treated with 20 μM S1 for 16 h. Mcl-1, Bcl-2 and pBcl-2 immunoprecipitations were performed, and immunoprecipitated fractions were analysed by Western blotting for the indicated proteins. (D) Phosphorylated Bcl-2 activates ERK1/2. Parental H1688 cells and H1688-expressing WT-Bcl-2, AAA-Bcl-2 or EEE-Bcl-2 were treated with and without 10 μM S1 or ABT-737 for 16 h and then lysed. Protein levels were examined by immunoblotting. Cell viability was assessed by MTS assay.
    Figure Legend Snippet: Bcl-2 is phosphorylated by ERK1/2 in H1688-derived cells and contributes to the resistance to S1. (A) PD98059 was able to block Bcl-2 phosphorylation and reduce the pBcl-2/Bcl-2 ratios. H1688-derived H1688-SR6, H1688-SR10 cell, H1048 and H740 cells were incubated with 5 μM PD98059 for 16 h. DMSO was used as a control. Total protein extracts (50 μg) from these cells were analysed by Western blot for pBcl-2 and Bcl-2 expression. The mean of pBcl-2/Bcl-2 ratio is shown for each cell line. (B) pBcl-2 sequestrated more pro-apoptotic proteins in acquired resistant cells. Immunoprecipitation of H1688-derived cell lysates. Mcl-1, Bcl-2 and pBcl-2 immunoprecipitations were performed, and immunoprecipitated fractions were analysed by Western blotting for the indicated proteins. (C) H1688-SR10 cells were treated with 20 μM S1 for 16 h. Mcl-1, Bcl-2 and pBcl-2 immunoprecipitations were performed, and immunoprecipitated fractions were analysed by Western blotting for the indicated proteins. (D) Phosphorylated Bcl-2 activates ERK1/2. Parental H1688 cells and H1688-expressing WT-Bcl-2, AAA-Bcl-2 or EEE-Bcl-2 were treated with and without 10 μM S1 or ABT-737 for 16 h and then lysed. Protein levels were examined by immunoblotting. Cell viability was assessed by MTS assay.

    Techniques Used: Derivative Assay, Blocking Assay, Incubation, Western Blot, Expressing, Immunoprecipitation, MTS Assay

    Schematic working model for ER stress/ERK1/2-mediated signalling of BH3 mimetics (e.g. S1)-induced Bcl-2 expression and phosphorylation. BH3 mimetics, such as S1 and ABT-737, stimulate ER stress then induce MAPK/ERK cascades with respect to Bcl-2 transcriptional up-regulation (solid arrowhead) and phosphorylation (hollow arrowhead). Excess pBcl-2 leads to cell survival by promoting ERK1/2 activation and sequestrating more pro-apoptotic members, which BH3 mimetics cannot release.
    Figure Legend Snippet: Schematic working model for ER stress/ERK1/2-mediated signalling of BH3 mimetics (e.g. S1)-induced Bcl-2 expression and phosphorylation. BH3 mimetics, such as S1 and ABT-737, stimulate ER stress then induce MAPK/ERK cascades with respect to Bcl-2 transcriptional up-regulation (solid arrowhead) and phosphorylation (hollow arrowhead). Excess pBcl-2 leads to cell survival by promoting ERK1/2 activation and sequestrating more pro-apoptotic members, which BH3 mimetics cannot release.

    Techniques Used: Expressing, Activation Assay

    erk1 2 sirna  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc bcl 2 sirna
    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the <t>Bcl-2</t> gene.
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    Cell Signaling Technology Inc signalsilence ampkα2 sirna ii
    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the <t>Bcl-2</t> gene.
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    Flow chart of the comprehensive screening for synthetic lethal genes interacting with <t>p53</t> R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.
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    Flow chart of the comprehensive screening for synthetic lethal genes interacting with <t>p53</t> R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.
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    Flow chart of the comprehensive screening for synthetic lethal genes interacting with <t>p53</t> R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.
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    Image Search Results


    T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.

    Journal: International Journal of Nanomedicine

    Article Title: An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma

    doi: 10.2147/IJN.S32900

    Figure Lengend Snippet: T 2 -weighted imaging of SK-N-SH cells. ( A ) T 2 -weighted imaging of SK-N-SH cells after 2-hour incubations with scAb GD2 -PEG-g-PEI-SPION, PEG-g-PEI-SPION, and scAb GD2 -PEG-g-PEI-SPION/GD2 at Fe concentrations of 5, 10, 20 and 40 μg/mL in DMEM. Untreated cells were used as the control group. Cells were scanned with a 1.5T MRI scanner. ( B ) shows the normalized MR intensity of different polymers at various Fe concentrations (n = 3). Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; DMEM, Dulbecco’s modified eagle medium; MRI, magnetic resonance imaging; MR, magnetic resonance. with either nontargeting or targeting/GD2 polyplexes. The results are consistent with results obtained through RT-PCR and indicate that targeting polyplexes have a stronger suppressive effect on the Bcl-2 gene.

    Article Snippet: FITC-siRNA, Bcl-2 siRNA, Rabbit anti-human Bcl-2 antibody and horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody were purchased from Cell Signaling Technology (CST, USA).

    Techniques: Imaging, Modification, Magnetic Resonance Imaging, Reverse Transcription Polymerase Chain Reaction

    Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.

    Journal: International Journal of Nanomedicine

    Article Title: An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma

    doi: 10.2147/IJN.S32900

    Figure Lengend Snippet: Efficiencies of PEG-g-PEI-SPION/siRNA, scAb GD2 -PEG-g-PEI-SPION/ siRNA, and scAb GD2 -PEG-g-PEI-SPION/siRNA with free GD2 antibody in suppressing Bcl-2 gene expression in SK-N-SH cells (N/P = 10). ( A ) Suppression of Bcl-2 mRNA levels as quantified by real-time RT-PCR (n = 3), When compared to scAb GD2 -PEG-g-PEI-SPION/siRNA (targeting), * P < 0.01 for control, nontargeting and targeting/GD2 polyplexes. ( B ) Suppression of the protein expression of the Bcl-2 gene was evaluated by Western blot analysis. Abbreviations: PEG-g-PEI-SPION, polyethylene glycol-grafted polyethylenimine superparamagnetic iron oxide nanoparticle; siRNA, small interfering ribonucleic acid; RT-PCR, reverse transcription polymerase chain reaction; P , probability level.

    Article Snippet: FITC-siRNA, Bcl-2 siRNA, Rabbit anti-human Bcl-2 antibody and horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody were purchased from Cell Signaling Technology (CST, USA).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription Polymerase Chain Reaction

    Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.

    Journal: International Journal of Nanomedicine

    Article Title: An MRI-visible non-viral vector for targeted Bcl-2 siRNA delivery to neuroblastoma

    doi: 10.2147/IJN.S32900

    Figure Lengend Snippet: Immunohistological characteristics of SK-N-SH tumors (sections were immunostained with Bcl-2 monoclonal antibodies). The brown stains indicated Bcl-2 protein. ( A and B ) PBS control, ( C and D ) nontargeting polyplex therapy, ( E and F ) targeting polyplex therapy. The black arrow mark apoptotic cell with condensation or nuclei fragmentation. Abbreviation: PBS, phosphate buffered saline.

    Article Snippet: FITC-siRNA, Bcl-2 siRNA, Rabbit anti-human Bcl-2 antibody and horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody were purchased from Cell Signaling Technology (CST, USA).

    Techniques:

    KEY RESOURCES TABLE

    Journal: Cell metabolism

    Article Title: The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress

    doi: 10.1016/j.cmet.2018.06.008

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: AMPKα2siRNAII , Cell Signaling Technology , Cat# 6630S.

    Techniques: Recombinant, SYBR Green Assay, Reporter Assay, Over Expression, Sequencing, Luciferase, Software, Real-time Polymerase Chain Reaction

    Flow chart of the comprehensive screening for synthetic lethal genes interacting with p53 R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.

    Journal: Oncology Reports

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    doi: 10.3892/or.2013.2953

    Figure Lengend Snippet: Flow chart of the comprehensive screening for synthetic lethal genes interacting with p53 R175H. Change in the abundance of a particular shRNA barcode is tracked for 10 days by competitive hybridization between the Dox-on and Dox-off groups. The cells in which shRNAs induced synthetic lethality with p53 R175H are selectively depleted from the Dox-on group. The same experiment was also conducted in SF126-tet-TON cells to exclude the possibility of doxycycline toxicity.

    Article Snippet: TP53 siRNA was purchased from Applied Biosystems (Foster City, CA, USA), and TP53-2 siRNA was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA).

    Techniques: shRNA, Hybridization

    Suppression of candidate genes in PC3 cells with p53 R175H expression. Ninety-six hours after transfection of plasmid pCR259 or pCR259-R175H in PC3 cells, (A) p53 R175H expression was confirmed by western blotting, and (B) cell numbers were measured by performing cell proliferation assays. The vertical axis corresponds to the absorbance ratio of (candidate gene siRNA transfected cells)/(negative control siRNA transfected cells) for 5 candidate genes. Values shown are means ± SD (n=3). * p<0.05 between p53 R175H null and p53 R175H expression. (C) Western blot analysis of Id1 and p53 R175H. Id1 expression was unchanged by R175H expression.

    Journal: Oncology Reports

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    doi: 10.3892/or.2013.2953

    Figure Lengend Snippet: Suppression of candidate genes in PC3 cells with p53 R175H expression. Ninety-six hours after transfection of plasmid pCR259 or pCR259-R175H in PC3 cells, (A) p53 R175H expression was confirmed by western blotting, and (B) cell numbers were measured by performing cell proliferation assays. The vertical axis corresponds to the absorbance ratio of (candidate gene siRNA transfected cells)/(negative control siRNA transfected cells) for 5 candidate genes. Values shown are means ± SD (n=3). * p<0.05 between p53 R175H null and p53 R175H expression. (C) Western blot analysis of Id1 and p53 R175H. Id1 expression was unchanged by R175H expression.

    Article Snippet: TP53 siRNA was purchased from Applied Biosystems (Foster City, CA, USA), and TP53-2 siRNA was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA).

    Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot, Negative Control

    Comparison of cell growth inhibition by ID1 suppression alone and ID1 / TP53 double suppression in cells expressing p53 R175H, wild-type p53, and TP53 -null cells. (A) ID1 and TP53 siRNA were co-transfected into SKBr3 cells to make a final concentration of 100 nM, and cell numbers were measured by performing cell proliferation assays on days 2 and 4. The vertical axis corresponds to the absorbance of the cell proliferation assay. Top of each figure represents expression inhibition of Id1 or p53 by each siRNA. Furthermore, other siRNAs of ID1 ( ID1 - 2 ) (B) and TP53 ( TP53 -2) (C) targeting different sites were transfected into SKBr3 cells. The same experiments were conducted in LS123 and HCC1395 cells, p53 R175H expressing cell lines (D and E), HCT116 cells expressing wild-type p53 (F), and the TP53 -null cell line PC3 (G). Values shown are means ± SD (n=3) in (A and E). * p<0.05, ** p<0.01, between ID1 suppression alone and ID1 / TP53 double suppression. (H) Western blot analysis of Id1 and p53 R175H. Knockdown level of Id1 was not rescued by ID1 / TP53 double suppression in SKBr3 cells.

    Journal: Oncology Reports

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    doi: 10.3892/or.2013.2953

    Figure Lengend Snippet: Comparison of cell growth inhibition by ID1 suppression alone and ID1 / TP53 double suppression in cells expressing p53 R175H, wild-type p53, and TP53 -null cells. (A) ID1 and TP53 siRNA were co-transfected into SKBr3 cells to make a final concentration of 100 nM, and cell numbers were measured by performing cell proliferation assays on days 2 and 4. The vertical axis corresponds to the absorbance of the cell proliferation assay. Top of each figure represents expression inhibition of Id1 or p53 by each siRNA. Furthermore, other siRNAs of ID1 ( ID1 - 2 ) (B) and TP53 ( TP53 -2) (C) targeting different sites were transfected into SKBr3 cells. The same experiments were conducted in LS123 and HCC1395 cells, p53 R175H expressing cell lines (D and E), HCT116 cells expressing wild-type p53 (F), and the TP53 -null cell line PC3 (G). Values shown are means ± SD (n=3) in (A and E). * p<0.05, ** p<0.01, between ID1 suppression alone and ID1 / TP53 double suppression. (H) Western blot analysis of Id1 and p53 R175H. Knockdown level of Id1 was not rescued by ID1 / TP53 double suppression in SKBr3 cells.

    Article Snippet: TP53 siRNA was purchased from Applied Biosystems (Foster City, CA, USA), and TP53-2 siRNA was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA).

    Techniques: Inhibition, Expressing, Transfection, Concentration Assay, Proliferation Assay, Western Blot

    ID1 suppression in PC3 cells expressing p53 R273H and ID1 / TP53 double suppression in cells expressing p53 R273H. (A) The plasmid pCR259-R273H was transfected into PC3 cells for 96 h, and total cell numbers were measured by performing cell proliferation assays. ID1 siRNA and TP53 siRNA were co-transfected into HT-29 cells expressing endogenous p53 R273H (B) and SW480 expressing endogenous p53 R273H/R309S double mutant (C). Cell counts were measured by performing cell proliferation assays on days 2 and 4. The vertical axis is same as in . The top right of each figure is the same as that in . Values shown are means ± SD (n=3).

    Journal: Oncology Reports

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    doi: 10.3892/or.2013.2953

    Figure Lengend Snippet: ID1 suppression in PC3 cells expressing p53 R273H and ID1 / TP53 double suppression in cells expressing p53 R273H. (A) The plasmid pCR259-R273H was transfected into PC3 cells for 96 h, and total cell numbers were measured by performing cell proliferation assays. ID1 siRNA and TP53 siRNA were co-transfected into HT-29 cells expressing endogenous p53 R273H (B) and SW480 expressing endogenous p53 R273H/R309S double mutant (C). Cell counts were measured by performing cell proliferation assays on days 2 and 4. The vertical axis is same as in . The top right of each figure is the same as that in . Values shown are means ± SD (n=3).

    Article Snippet: TP53 siRNA was purchased from Applied Biosystems (Foster City, CA, USA), and TP53-2 siRNA was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA).

    Techniques: Expressing, Plasmid Preparation, Transfection, Mutagenesis

    FACS analysis of SKBr3 (A), HCT116 (B) and PC3 (C) cells with ID1 and TP53 double suppression. FACS analysis with or without ID1 siRNA (100 nM) and TP53 siRNA (100 nM). Values of the sub-G1 fraction are a portion of the total population, and values of G1, S, and G2/M fractions are portions of the population excluding the sub-G1 fraction. Values shown are means ± SD (n=6). * p<0.05, ** p<0.01, between negative control and ID1 suppression or between ID1 suppression and ID1 / TP53 double suppression.

    Journal: Oncology Reports

    Article Title: High throughput RNAi screening identifies ID1 as a synthetic sick/lethal gene interacting with the common TP53 mutation R175H

    doi: 10.3892/or.2013.2953

    Figure Lengend Snippet: FACS analysis of SKBr3 (A), HCT116 (B) and PC3 (C) cells with ID1 and TP53 double suppression. FACS analysis with or without ID1 siRNA (100 nM) and TP53 siRNA (100 nM). Values of the sub-G1 fraction are a portion of the total population, and values of G1, S, and G2/M fractions are portions of the population excluding the sub-G1 fraction. Values shown are means ± SD (n=6). * p<0.05, ** p<0.01, between negative control and ID1 suppression or between ID1 suppression and ID1 / TP53 double suppression.

    Article Snippet: TP53 siRNA was purchased from Applied Biosystems (Foster City, CA, USA), and TP53-2 siRNA was purchased from Cell Signaling Technology, Inc. (Boston, MA, USA).

    Techniques: Negative Control