u0126  (Alomone Labs)


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    Structured Review

    Alomone Labs u0126
    IMP3 exerts its proproliferative effects through the IGF-2-PI3K and IGF-2-MAPK cascades. A , viability was measured by MTT assay at indicated time points for H1299 vector and IMP3 stable clones treated with a neutralizing antibody for IGF-2 as indicated. The assays were carried out in triplicate, and the mean value for each cell type at each time point was used to generate the graph. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups and a p ≤ 0.05 is represented with an asterisk , p ≤ 0.01 is represented as double asterisks and p ≤ 0.0001 is represented as triple asterisks. B , viability was measured by MTT assay as an indicator of cell proliferation at the indicated time points for either mock, cyclophilin ( cyclo. ), or IMP3 siRNA-transfected U138 cells. IGF-2 was exogenously supplemented as indicated to monitor its ability to rescue the effects of IMP3 knockdown. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups. A p value ≤ 0.05 is represented as an asterisk , which is significant, and NS refers to nonsignificant difference. C , equal amounts of total protein lysates from cyclophilin and IMP3 siRNA-treated U138 cells were subjected to Western blotting with the indicated antibodies. Note that the intensity of phospho-mTOR, phospho-Akt, phospho-4EBP1, phospho-MEK1/2, and phospho-ERK1/2 bands reduces in lane 2 compared with lane 1 , whereas that of total the corresponding total proteins remain unchanged. D , viability was measured by MTT assay on day 9 after plating for the U373 vector clone (#6) and IMP3 stable clones (#5 and 17) treated with dimethyl sulfoxide (vehicle), LY294002 (20 μ m ), or <t>U0126</t> (10 μ m ). Please note that dimethyl sulfoxide ( DMSO )-treated IMP3 stable clone (#5 and #17) grow faster than vector stable clone (compare light gray bars ). Both LY294002 and U0126 treated IMP3 stable clone (#5 and #17) failed to show faster growth than vector stable clone (compare dark gray and black bars with light gray bars ).
    U0126, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/u0126/product/Alomone Labs
    Average 92 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    u0126 - by Bioz Stars, 2022-11
    92/100 stars

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    1) Product Images from "Insulin Growth Factor-2 Binding Protein 3 (IGF2BP3) Is a Glioblastoma-specific Marker That Activates Phosphatidylinositol 3-Kinase/Mitogen-activated Protein Kinase (PI3K/MAPK) Pathways by Modulating IGF-2 *"

    Article Title: Insulin Growth Factor-2 Binding Protein 3 (IGF2BP3) Is a Glioblastoma-specific Marker That Activates Phosphatidylinositol 3-Kinase/Mitogen-activated Protein Kinase (PI3K/MAPK) Pathways by Modulating IGF-2 *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.178012

    IMP3 exerts its proproliferative effects through the IGF-2-PI3K and IGF-2-MAPK cascades. A , viability was measured by MTT assay at indicated time points for H1299 vector and IMP3 stable clones treated with a neutralizing antibody for IGF-2 as indicated. The assays were carried out in triplicate, and the mean value for each cell type at each time point was used to generate the graph. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups and a p ≤ 0.05 is represented with an asterisk , p ≤ 0.01 is represented as double asterisks and p ≤ 0.0001 is represented as triple asterisks. B , viability was measured by MTT assay as an indicator of cell proliferation at the indicated time points for either mock, cyclophilin ( cyclo. ), or IMP3 siRNA-transfected U138 cells. IGF-2 was exogenously supplemented as indicated to monitor its ability to rescue the effects of IMP3 knockdown. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups. A p value ≤ 0.05 is represented as an asterisk , which is significant, and NS refers to nonsignificant difference. C , equal amounts of total protein lysates from cyclophilin and IMP3 siRNA-treated U138 cells were subjected to Western blotting with the indicated antibodies. Note that the intensity of phospho-mTOR, phospho-Akt, phospho-4EBP1, phospho-MEK1/2, and phospho-ERK1/2 bands reduces in lane 2 compared with lane 1 , whereas that of total the corresponding total proteins remain unchanged. D , viability was measured by MTT assay on day 9 after plating for the U373 vector clone (#6) and IMP3 stable clones (#5 and 17) treated with dimethyl sulfoxide (vehicle), LY294002 (20 μ m ), or U0126 (10 μ m ). Please note that dimethyl sulfoxide ( DMSO )-treated IMP3 stable clone (#5 and #17) grow faster than vector stable clone (compare light gray bars ). Both LY294002 and U0126 treated IMP3 stable clone (#5 and #17) failed to show faster growth than vector stable clone (compare dark gray and black bars with light gray bars ).
    Figure Legend Snippet: IMP3 exerts its proproliferative effects through the IGF-2-PI3K and IGF-2-MAPK cascades. A , viability was measured by MTT assay at indicated time points for H1299 vector and IMP3 stable clones treated with a neutralizing antibody for IGF-2 as indicated. The assays were carried out in triplicate, and the mean value for each cell type at each time point was used to generate the graph. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups and a p ≤ 0.05 is represented with an asterisk , p ≤ 0.01 is represented as double asterisks and p ≤ 0.0001 is represented as triple asterisks. B , viability was measured by MTT assay as an indicator of cell proliferation at the indicated time points for either mock, cyclophilin ( cyclo. ), or IMP3 siRNA-transfected U138 cells. IGF-2 was exogenously supplemented as indicated to monitor its ability to rescue the effects of IMP3 knockdown. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups. A p value ≤ 0.05 is represented as an asterisk , which is significant, and NS refers to nonsignificant difference. C , equal amounts of total protein lysates from cyclophilin and IMP3 siRNA-treated U138 cells were subjected to Western blotting with the indicated antibodies. Note that the intensity of phospho-mTOR, phospho-Akt, phospho-4EBP1, phospho-MEK1/2, and phospho-ERK1/2 bands reduces in lane 2 compared with lane 1 , whereas that of total the corresponding total proteins remain unchanged. D , viability was measured by MTT assay on day 9 after plating for the U373 vector clone (#6) and IMP3 stable clones (#5 and 17) treated with dimethyl sulfoxide (vehicle), LY294002 (20 μ m ), or U0126 (10 μ m ). Please note that dimethyl sulfoxide ( DMSO )-treated IMP3 stable clone (#5 and #17) grow faster than vector stable clone (compare light gray bars ). Both LY294002 and U0126 treated IMP3 stable clone (#5 and #17) failed to show faster growth than vector stable clone (compare dark gray and black bars with light gray bars ).

    Techniques Used: MTT Assay, Plasmid Preparation, Clone Assay, Transfection, Western Blot, Stable Transfection

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    Alomone Labs mapkk mek1 2 inhibitor u0126
    Il-1β secretion, Mitochondrial NADH cytochrome c reductase activity and ROS levels in stable CFTR knock down cells. Caco-2/pRSctrl cells (transfected with pRS control) and Caco-2/pRS26 cells (transfected with the shRNA pRS26) were preincubated 24 h in serum free media. A: Immune-dot blotting quantification of the IL-1β present in culture media. The results were expressed as pg/ml. B: Cells were treated for additional 24 h, as indicated. IL-1β: IL-1β (5 ng/ml); Ab-IL-1β: anti-IL-1β blocking antibody (30 ng/ml); Ab-Ctrl: anti-Histone H1 monoclonal antibody as negative control (30 ng/ml); p38 Inh: p38 inhibitor SB203580 (5 µM); IKK Inh: IKK inhibitor III (2 µM); JNK Inh: JNK inhibitor SP600125 (5 µM); <t>MEK1/2</t> Inh: MAPK/AP-1 pathway inhibitor U0126 (5 µM). Spectrophotometric measurements of the mitochondrial NADH-cytochrome c reductase activity, expressed as percentage (%) relative to Caco-2/pRS26 control values. C: Mitochondrial ROS levels in the same experiments of panel B; the figure shows the MitoSOX fluorescence (5 µM for 10 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. D: Cellular ROS levels in the same experiments of panel B; the figure shows the DCF fluorescence (DCFH-DA 10 µM for 40 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. Measurements were performed in triplicate and data are expressed as mean ± SE of two independent experiments (n = 2). *indicates p
    Mapkk Mek1 2 Inhibitor U0126, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mapkk mek1 2 inhibitor u0126/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mapkk mek1 2 inhibitor u0126 - by Bioz Stars, 2022-11
    92/100 stars
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    Il-1β secretion, Mitochondrial NADH cytochrome c reductase activity and ROS levels in stable CFTR knock down cells. Caco-2/pRSctrl cells (transfected with pRS control) and Caco-2/pRS26 cells (transfected with the shRNA pRS26) were preincubated 24 h in serum free media. A: Immune-dot blotting quantification of the IL-1β present in culture media. The results were expressed as pg/ml. B: Cells were treated for additional 24 h, as indicated. IL-1β: IL-1β (5 ng/ml); Ab-IL-1β: anti-IL-1β blocking antibody (30 ng/ml); Ab-Ctrl: anti-Histone H1 monoclonal antibody as negative control (30 ng/ml); p38 Inh: p38 inhibitor SB203580 (5 µM); IKK Inh: IKK inhibitor III (2 µM); JNK Inh: JNK inhibitor SP600125 (5 µM); MEK1/2 Inh: MAPK/AP-1 pathway inhibitor U0126 (5 µM). Spectrophotometric measurements of the mitochondrial NADH-cytochrome c reductase activity, expressed as percentage (%) relative to Caco-2/pRS26 control values. C: Mitochondrial ROS levels in the same experiments of panel B; the figure shows the MitoSOX fluorescence (5 µM for 10 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. D: Cellular ROS levels in the same experiments of panel B; the figure shows the DCF fluorescence (DCFH-DA 10 µM for 40 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. Measurements were performed in triplicate and data are expressed as mean ± SE of two independent experiments (n = 2). *indicates p

    Journal: PLoS ONE

    Article Title: Disruption of Interleukin-1? Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    doi: 10.1371/journal.pone.0099257

    Figure Lengend Snippet: Il-1β secretion, Mitochondrial NADH cytochrome c reductase activity and ROS levels in stable CFTR knock down cells. Caco-2/pRSctrl cells (transfected with pRS control) and Caco-2/pRS26 cells (transfected with the shRNA pRS26) were preincubated 24 h in serum free media. A: Immune-dot blotting quantification of the IL-1β present in culture media. The results were expressed as pg/ml. B: Cells were treated for additional 24 h, as indicated. IL-1β: IL-1β (5 ng/ml); Ab-IL-1β: anti-IL-1β blocking antibody (30 ng/ml); Ab-Ctrl: anti-Histone H1 monoclonal antibody as negative control (30 ng/ml); p38 Inh: p38 inhibitor SB203580 (5 µM); IKK Inh: IKK inhibitor III (2 µM); JNK Inh: JNK inhibitor SP600125 (5 µM); MEK1/2 Inh: MAPK/AP-1 pathway inhibitor U0126 (5 µM). Spectrophotometric measurements of the mitochondrial NADH-cytochrome c reductase activity, expressed as percentage (%) relative to Caco-2/pRS26 control values. C: Mitochondrial ROS levels in the same experiments of panel B; the figure shows the MitoSOX fluorescence (5 µM for 10 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. D: Cellular ROS levels in the same experiments of panel B; the figure shows the DCF fluorescence (DCFH-DA 10 µM for 40 min in Hank’s buffer). Results were expressed as percentage (%) relative to Caco-2/pRSctrl control values. Measurements were performed in triplicate and data are expressed as mean ± SE of two independent experiments (n = 2). *indicates p

    Article Snippet: MAPK1/p38 inhibitor (SB203580), MAPKK (MEK1/2) inhibitor (U0126) and JNK inhibitor (SP600125) were from Alomone Labs (Jerusalem, Israel).

    Techniques: Activity Assay, Transfection, shRNA, Blocking Assay, Negative Control, Fluorescence

    MEK1/2 inhibition. After 24-1 cells were incubated for 24 h with increasing concentrations of the MAPK/AP-1 pathway inhibitor U0126 (1, 5, 10 and 20 µM) and the mCx-I activity was measured by using BN-PAGE and spectrophotometry. A: mCx-I in-gel activity (IGA) and mCx-III Western blotting (WB). B: Densitometric quantification and statistical analysis of the results shown in panel A, calculated from three independent experiments (n = 3). IGA of mCx-I was calculated as the ratio mCx-I (IGA)/mCx-III (WB). C: Spectrophotometric measurements of the mitochondrial NADH-cytochrome c reductase activity in the same experiments of panel A, expressed as percentage (%) relative to S9 control values. Measurements were performed in triplicate and data were expressed as mean ± SE of three independent experiments (n = 3). *indicates p

    Journal: PLoS ONE

    Article Title: Disruption of Interleukin-1? Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    doi: 10.1371/journal.pone.0099257

    Figure Lengend Snippet: MEK1/2 inhibition. After 24-1 cells were incubated for 24 h with increasing concentrations of the MAPK/AP-1 pathway inhibitor U0126 (1, 5, 10 and 20 µM) and the mCx-I activity was measured by using BN-PAGE and spectrophotometry. A: mCx-I in-gel activity (IGA) and mCx-III Western blotting (WB). B: Densitometric quantification and statistical analysis of the results shown in panel A, calculated from three independent experiments (n = 3). IGA of mCx-I was calculated as the ratio mCx-I (IGA)/mCx-III (WB). C: Spectrophotometric measurements of the mitochondrial NADH-cytochrome c reductase activity in the same experiments of panel A, expressed as percentage (%) relative to S9 control values. Measurements were performed in triplicate and data were expressed as mean ± SE of three independent experiments (n = 3). *indicates p

    Article Snippet: MAPK1/p38 inhibitor (SB203580), MAPKK (MEK1/2) inhibitor (U0126) and JNK inhibitor (SP600125) were from Alomone Labs (Jerusalem, Israel).

    Techniques: Inhibition, Incubation, Activity Assay, Polyacrylamide Gel Electrophoresis, Spectrophotometry, Western Blot

    ROS levels in IB3-1 and S9 cells. S9 and IB3-1 cells were preincubated 24 h in serum free media. Then, the media was replaced (serum free, in the presence of a CFTR-stimulating cocktail) and the cells incubated another 24 h. The IB3-1 cells were treated in this second incubation as indicated. IL-1β: IL-1β (5 ng/ml); Ab-IL-1β: anti-IL-1β blocking antibody (30 ng/ml); Ab-Ctrl: anti-Histone H1 monoclonal antibody as negative control for antibody incubation (30 ng/ml); p38 Inh: p38 inhibitor SB203580 (5 µM); IKK Inh: IKK inhibitor III (2 µM); JNK Inh: JNK inhibitor SP600125 (5 µM); MEK1/2 Inh: MAPK/AP-1 pathway inhibitor U0126 (5 µM). A: MitoSOX fluorescence (5 µM for 10 min in Hank’s buffer). B: DCF fluorescence (10 µM DCFH-DA for 40 min in Hank’s buffer); Ab-Ctrl: anti-JNK2 monoclonal antibody as negative control. Results were expressed as percentage (%) relative to S9 control values. Measurements were performed in triplicate and data are expressed as mean ± SE of three independent experiments (n = 3). *indicates p

    Journal: PLoS ONE

    Article Title: Disruption of Interleukin-1? Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    doi: 10.1371/journal.pone.0099257

    Figure Lengend Snippet: ROS levels in IB3-1 and S9 cells. S9 and IB3-1 cells were preincubated 24 h in serum free media. Then, the media was replaced (serum free, in the presence of a CFTR-stimulating cocktail) and the cells incubated another 24 h. The IB3-1 cells were treated in this second incubation as indicated. IL-1β: IL-1β (5 ng/ml); Ab-IL-1β: anti-IL-1β blocking antibody (30 ng/ml); Ab-Ctrl: anti-Histone H1 monoclonal antibody as negative control for antibody incubation (30 ng/ml); p38 Inh: p38 inhibitor SB203580 (5 µM); IKK Inh: IKK inhibitor III (2 µM); JNK Inh: JNK inhibitor SP600125 (5 µM); MEK1/2 Inh: MAPK/AP-1 pathway inhibitor U0126 (5 µM). A: MitoSOX fluorescence (5 µM for 10 min in Hank’s buffer). B: DCF fluorescence (10 µM DCFH-DA for 40 min in Hank’s buffer); Ab-Ctrl: anti-JNK2 monoclonal antibody as negative control. Results were expressed as percentage (%) relative to S9 control values. Measurements were performed in triplicate and data are expressed as mean ± SE of three independent experiments (n = 3). *indicates p

    Article Snippet: MAPK1/p38 inhibitor (SB203580), MAPKK (MEK1/2) inhibitor (U0126) and JNK inhibitor (SP600125) were from Alomone Labs (Jerusalem, Israel).

    Techniques: Incubation, Blocking Assay, Negative Control, Fluorescence

    Graphical summary for IL-1β effects on mCx-I activity and mitochondrial ROS levels in IB3-1 and Caco-2/pRS26. The figure illustrates the interactions among the different proteins, kinases or small molecules involved in this work. The interactions were drawn by using the software Pathway Studio (v 9, Elsevier). Arrows with the+symbol represent stimulations and those with the -| symbol represent inhibition. Green ellipses: small molecules; red sickle-vertex: kinases; purple rectangle: disease (CF); blue star-vertex: shRNA specific for CFTR. The results obtained with IL-1β blocking Ab or with the receptor inhibitor IL1RN suggest that an autocrine IL-1β signaling is responsible for the reduced mCx-I activity and the increased ROS levels seen in IB3-1 CF cells or in Caco-2/pRS26 cells. Inhibition of NF-κB or p38 MAPK also resulted in increased mCx-I activity and decreased ROS levels. The inhibition of MEK1/2 or JNKs (AP-1 pathway) had no effects. The mechanisms by which CFTR increases IL-1β and IL1-β, p38 MAPKs or NF-κB inhibit mCx-I and increase ROS levels remain to be determined (dotted lines).

    Journal: PLoS ONE

    Article Title: Disruption of Interleukin-1? Autocrine Signaling Rescues Complex I Activity and Improves ROS Levels in Immortalized Epithelial Cells with Impaired Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function

    doi: 10.1371/journal.pone.0099257

    Figure Lengend Snippet: Graphical summary for IL-1β effects on mCx-I activity and mitochondrial ROS levels in IB3-1 and Caco-2/pRS26. The figure illustrates the interactions among the different proteins, kinases or small molecules involved in this work. The interactions were drawn by using the software Pathway Studio (v 9, Elsevier). Arrows with the+symbol represent stimulations and those with the -| symbol represent inhibition. Green ellipses: small molecules; red sickle-vertex: kinases; purple rectangle: disease (CF); blue star-vertex: shRNA specific for CFTR. The results obtained with IL-1β blocking Ab or with the receptor inhibitor IL1RN suggest that an autocrine IL-1β signaling is responsible for the reduced mCx-I activity and the increased ROS levels seen in IB3-1 CF cells or in Caco-2/pRS26 cells. Inhibition of NF-κB or p38 MAPK also resulted in increased mCx-I activity and decreased ROS levels. The inhibition of MEK1/2 or JNKs (AP-1 pathway) had no effects. The mechanisms by which CFTR increases IL-1β and IL1-β, p38 MAPKs or NF-κB inhibit mCx-I and increase ROS levels remain to be determined (dotted lines).

    Article Snippet: MAPK1/p38 inhibitor (SB203580), MAPKK (MEK1/2) inhibitor (U0126) and JNK inhibitor (SP600125) were from Alomone Labs (Jerusalem, Israel).

    Techniques: Activity Assay, Software, Inhibition, shRNA, Blocking Assay

    IMP3 exerts its proproliferative effects through the IGF-2-PI3K and IGF-2-MAPK cascades. A , viability was measured by MTT assay at indicated time points for H1299 vector and IMP3 stable clones treated with a neutralizing antibody for IGF-2 as indicated. The assays were carried out in triplicate, and the mean value for each cell type at each time point was used to generate the graph. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups and a p ≤ 0.05 is represented with an asterisk , p ≤ 0.01 is represented as double asterisks and p ≤ 0.0001 is represented as triple asterisks. B , viability was measured by MTT assay as an indicator of cell proliferation at the indicated time points for either mock, cyclophilin ( cyclo. ), or IMP3 siRNA-transfected U138 cells. IGF-2 was exogenously supplemented as indicated to monitor its ability to rescue the effects of IMP3 knockdown. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups. A p value ≤ 0.05 is represented as an asterisk , which is significant, and NS refers to nonsignificant difference. C , equal amounts of total protein lysates from cyclophilin and IMP3 siRNA-treated U138 cells were subjected to Western blotting with the indicated antibodies. Note that the intensity of phospho-mTOR, phospho-Akt, phospho-4EBP1, phospho-MEK1/2, and phospho-ERK1/2 bands reduces in lane 2 compared with lane 1 , whereas that of total the corresponding total proteins remain unchanged. D , viability was measured by MTT assay on day 9 after plating for the U373 vector clone (#6) and IMP3 stable clones (#5 and 17) treated with dimethyl sulfoxide (vehicle), LY294002 (20 μ m ), or U0126 (10 μ m ). Please note that dimethyl sulfoxide ( DMSO )-treated IMP3 stable clone (#5 and #17) grow faster than vector stable clone (compare light gray bars ). Both LY294002 and U0126 treated IMP3 stable clone (#5 and #17) failed to show faster growth than vector stable clone (compare dark gray and black bars with light gray bars ).

    Journal: The Journal of Biological Chemistry

    Article Title: Insulin Growth Factor-2 Binding Protein 3 (IGF2BP3) Is a Glioblastoma-specific Marker That Activates Phosphatidylinositol 3-Kinase/Mitogen-activated Protein Kinase (PI3K/MAPK) Pathways by Modulating IGF-2 *

    doi: 10.1074/jbc.M110.178012

    Figure Lengend Snippet: IMP3 exerts its proproliferative effects through the IGF-2-PI3K and IGF-2-MAPK cascades. A , viability was measured by MTT assay at indicated time points for H1299 vector and IMP3 stable clones treated with a neutralizing antibody for IGF-2 as indicated. The assays were carried out in triplicate, and the mean value for each cell type at each time point was used to generate the graph. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups and a p ≤ 0.05 is represented with an asterisk , p ≤ 0.01 is represented as double asterisks and p ≤ 0.0001 is represented as triple asterisks. B , viability was measured by MTT assay as an indicator of cell proliferation at the indicated time points for either mock, cyclophilin ( cyclo. ), or IMP3 siRNA-transfected U138 cells. IGF-2 was exogenously supplemented as indicated to monitor its ability to rescue the effects of IMP3 knockdown. A one-way analysis of variance was carried out to test the significance of the observed differences between the groups. A p value ≤ 0.05 is represented as an asterisk , which is significant, and NS refers to nonsignificant difference. C , equal amounts of total protein lysates from cyclophilin and IMP3 siRNA-treated U138 cells were subjected to Western blotting with the indicated antibodies. Note that the intensity of phospho-mTOR, phospho-Akt, phospho-4EBP1, phospho-MEK1/2, and phospho-ERK1/2 bands reduces in lane 2 compared with lane 1 , whereas that of total the corresponding total proteins remain unchanged. D , viability was measured by MTT assay on day 9 after plating for the U373 vector clone (#6) and IMP3 stable clones (#5 and 17) treated with dimethyl sulfoxide (vehicle), LY294002 (20 μ m ), or U0126 (10 μ m ). Please note that dimethyl sulfoxide ( DMSO )-treated IMP3 stable clone (#5 and #17) grow faster than vector stable clone (compare light gray bars ). Both LY294002 and U0126 treated IMP3 stable clone (#5 and #17) failed to show faster growth than vector stable clone (compare dark gray and black bars with light gray bars ).

    Article Snippet: LY294002 and U0126, the pharmacological inhibitors for the PI3K and the MAPK pathways were obtained from Alomone Biosciences.

    Techniques: MTT Assay, Plasmid Preparation, Clone Assay, Transfection, Western Blot, Stable Transfection