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constitutive active map2k4  (Addgene inc)


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    Addgene inc constitutive active map2k4
    A ) <t>MAP2K4</t> protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.
    Constitutive Active Map2k4, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/constitutive+active+map2k4/pCDNA3+Flag+MKK4+(Plasmid+%2314615)/pmc04096757-62-45-51
    Average 93 stars, based on 6 article reviews
    constitutive active map2k4 - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis"

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102289

    A ) MAP2K4 protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.
    Figure Legend Snippet: A ) MAP2K4 protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.

    Techniques Used: Expressing, Variant Assay, Plasmid Preparation, Control, Western Blot

    Studies were conducted on MAP2K4 variant cell lines. A ) Knockdown of MAP2K4 protein by siRNA. After transfection of cells with siRNA targeting MAP2K4 (siMAP2K4) or non-targeting control (siCON), MAP2K4 expression was measured by Western blot. Percent knockdown is denoted above the blot. B ) Effect of MAP2K4 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Effects on cell migration. Cell migration was measured as described in . Data from individual types of clones were combined, are from three independent experiments, each in replicates of N = 3, and are expressed as the percentage of migrating cells, normalized to VC. D ) MMP transcript expression. The levels of MMP-2, MMP-9, MMP-10 and MMP-13 transcript levels were measured by qRT/PCR, normalized to GAPDH, and expressed as the percentage of VC. Data are from four independent experiments, each in replicates of N = 2. E ) Cell growth. Cells were plated at the indicated concentrations, allowed to grow for 5 days, MTT added, and optical density determined. Data are from three independent experiments, each N = 3. F ) Colony formation. Colony formation after 14 days was determined in three independent experiments, each N = 2, and expressed as the percent of VC. Values in all graphs are the mean ± SEM. * denotes p≤0.05 between the indicated groups.
    Figure Legend Snippet: Studies were conducted on MAP2K4 variant cell lines. A ) Knockdown of MAP2K4 protein by siRNA. After transfection of cells with siRNA targeting MAP2K4 (siMAP2K4) or non-targeting control (siCON), MAP2K4 expression was measured by Western blot. Percent knockdown is denoted above the blot. B ) Effect of MAP2K4 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Effects on cell migration. Cell migration was measured as described in . Data from individual types of clones were combined, are from three independent experiments, each in replicates of N = 3, and are expressed as the percentage of migrating cells, normalized to VC. D ) MMP transcript expression. The levels of MMP-2, MMP-9, MMP-10 and MMP-13 transcript levels were measured by qRT/PCR, normalized to GAPDH, and expressed as the percentage of VC. Data are from four independent experiments, each in replicates of N = 2. E ) Cell growth. Cells were plated at the indicated concentrations, allowed to grow for 5 days, MTT added, and optical density determined. Data are from three independent experiments, each N = 3. F ) Colony formation. Colony formation after 14 days was determined in three independent experiments, each N = 2, and expressed as the percent of VC. Values in all graphs are the mean ± SEM. * denotes p≤0.05 between the indicated groups.

    Techniques Used: Variant Assay, Knockdown, Transfection, Control, Expressing, Western Blot, Migration, Clone Assay, Quantitative RT-PCR

    A ) MAP2K4 protein expression in LNCaP and 1542CPTX cell lines by Western Blot. B ) Relative cellular invasion of LNCaP and 1542CPTX (42C) cell lines as measured by a Matrigel coated Boyden chamber. Data are from four independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. C ) Relative MMP-2 mRNA expression in LNCaP and 42C cell lines as measured by qRT/PCR. Data are from five independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. D ) Phosphorylated and total HSP27 expression in LNCaP and 1542CPTX cell lines by Western Blot.
    Figure Legend Snippet: A ) MAP2K4 protein expression in LNCaP and 1542CPTX cell lines by Western Blot. B ) Relative cellular invasion of LNCaP and 1542CPTX (42C) cell lines as measured by a Matrigel coated Boyden chamber. Data are from four independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. C ) Relative MMP-2 mRNA expression in LNCaP and 42C cell lines as measured by qRT/PCR. Data are from five independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. D ) Phosphorylated and total HSP27 expression in LNCaP and 1542CPTX cell lines by Western Blot.

    Techniques Used: Expressing, Western Blot, Quantitative RT-PCR

    A ) Knockdown of HSP27 protein by siRNA. After transfection of cells with siRNA targeting HSP27 (siHSP27) or non-targeting control (siCON), HSP27 expression was measured by Western blot. B ) Effect of HSP27 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Knockdown of MMP-2 mRNA by siRNA. After transfection of cells with siRNA targeting MMP-2 (siMMP2) or non-targeting control (siCON), MMP-2 expression was measured by qRT/PCR. D ) Effect of MMP-2 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells.
    Figure Legend Snippet: A ) Knockdown of HSP27 protein by siRNA. After transfection of cells with siRNA targeting HSP27 (siHSP27) or non-targeting control (siCON), HSP27 expression was measured by Western blot. B ) Effect of HSP27 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Knockdown of MMP-2 mRNA by siRNA. After transfection of cells with siRNA targeting MMP-2 (siMMP2) or non-targeting control (siCON), MMP-2 expression was measured by qRT/PCR. D ) Effect of MMP-2 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells.

    Techniques Used: Knockdown, Transfection, Control, Expressing, Western Blot, Variant Assay, Quantitative RT-PCR

    Related Articles

    Negative Control:

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis
    Article Snippet: For knockdown experiments, Dharmacon ON-Targetplus SMARTpool siRNA targeting MAP2K4 (L-003574-00-0005), HSP27 (L-005269-00-0005), MMP-2 (L-005959-00-0005), or non-targeting siRNA (D-001810-10-05) was transfected into cells using Dharmafect Duo (T-2010, all from Thermo Scientific), along with pCMV-β-Galactosidase (Agilent Technologies), and used after 48 hours, as described by us . .. The following were purchased: antibodies, SEK1/MKK4 (#9152), phospho-p38 MAPK (T180/Y182) (#4631), p38 MAPK (#9212), phospho-SAPK/JNK (T183/Y185) (#4668), SAPK/JNK (#9258), phospho-HSP27 (#2401), HSP27 (#2402), and GAPDH (#2118), all from Cell Signaling Technology, and horseradish peroxidase-conjugated anti-mouse and anti-rabbit, from GE Healthcare Biosciences; plasmids, wild type and constitutive active MAP2K4 (#14615 and #14813; Addgene) and GFP (Vivid Colors pcDNA 6.2/N-EmGFP-GW/TOPO, Invitrogen); chemical inhibitors, p38 MAPK inhibitor, SB203580 (Sigma-Aldrich) and associated negative control, SB202474, and JNK Inhibitor II (SP600125), and JNK Inhibitor II Negative Control (N 1 -Methyl-1,9-pyrazoloanthrone), all from EMD Millipore. ..



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    constitutive active map2k4  (Addgene inc)
    93
    Addgene inc constitutive active map2k4
    A ) <t>MAP2K4</t> protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.
    Constitutive Active Map2k4, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/constitutive+active+map2k4/pCDNA3+Flag+MKK4+(Plasmid+%2314615)/pmc04096757-62-45-51
    Average 93 stars, based on 1 article reviews
    constitutive active map2k4 - by Bioz Stars, 2026-09
    93/100 stars
    		  Buy from Supplier

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    A ) MAP2K4 protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.

    Journal: PLoS ONE

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis

    doi: 10.1371/journal.pone.0102289

    Figure Lengend Snippet: A ) MAP2K4 protein expression in MAP2K4 variant cell lines. The expression of MAP2K4 protein was measured in wild type, WT1, WT2, and constitutive active, CA1, CA2, CA3, MAP2K4 over expressing cell lines, and vector control, VC1, VC2, cell lines by Western blot. A representative Western blot is depicted, with quantification of the MAP2K4/GAPDH levels, normalized to VC1 cells, shown above each lane. B ) Invasion of MAP2K4 variant cell lines. Boyden chamber Matrigel cell invasion assays were performed, as described in . Data are from 5 independent experiments, each in replicates of N = 3. Data from the respective wild type, constitutive active and vector control cell lines were combined to give an average read out for WT-MAP2K4, CA-MAP2K4 and VC cells, respectively. Data are expressed as the mean ± SEM percentage of VC cells. C-E ) The effect of MAP2K4 expression upon metastatic spread and tumor growth. Equal numbers of mice were orthotopically implanted with VC1 or VC2 cells (forming the VC cohort), WT1 or WT2 cells (WT cohort), or with CA1, CA2 or CA3 cells (CA cohort). In (C) the resultant number of distant metastasis is expressed as the mean ± SEM percentage of VC mice, in (E) the number of mice developing circulating tumor cells is depicted, and in (F) tumor volume is graphed as the mean ± SEM percentage of VC mice. A representative image of a distinct metastasis is shown in (D). * denotes p≤0.05 between VC and an experimental group.

    Article Snippet: The following were purchased: antibodies, SEK1/MKK4 (#9152), phospho-p38 MAPK (T180/Y182) (#4631), p38 MAPK (#9212), phospho-SAPK/JNK (T183/Y185) (#4668), SAPK/JNK (#9258), phospho-HSP27 (#2401), HSP27 (#2402), and GAPDH (#2118), all from Cell Signaling Technology, and horseradish peroxidase-conjugated anti-mouse and anti-rabbit, from GE Healthcare Biosciences; plasmids, wild type and constitutive active MAP2K4 (#14615 and #14813; Addgene) and GFP (Vivid Colors pcDNA 6.2/N-EmGFP-GW/TOPO, Invitrogen); chemical inhibitors, p38 MAPK inhibitor, SB203580 (Sigma-Aldrich) and associated negative control, SB202474, and JNK Inhibitor II (SP600125), and JNK Inhibitor II Negative Control (N 1 -Methyl-1,9-pyrazoloanthrone), all from EMD Millipore.

    Techniques: Expressing, Variant Assay, Plasmid Preparation, Control, Western Blot

    Studies were conducted on MAP2K4 variant cell lines. A ) Knockdown of MAP2K4 protein by siRNA. After transfection of cells with siRNA targeting MAP2K4 (siMAP2K4) or non-targeting control (siCON), MAP2K4 expression was measured by Western blot. Percent knockdown is denoted above the blot. B ) Effect of MAP2K4 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Effects on cell migration. Cell migration was measured as described in . Data from individual types of clones were combined, are from three independent experiments, each in replicates of N = 3, and are expressed as the percentage of migrating cells, normalized to VC. D ) MMP transcript expression. The levels of MMP-2, MMP-9, MMP-10 and MMP-13 transcript levels were measured by qRT/PCR, normalized to GAPDH, and expressed as the percentage of VC. Data are from four independent experiments, each in replicates of N = 2. E ) Cell growth. Cells were plated at the indicated concentrations, allowed to grow for 5 days, MTT added, and optical density determined. Data are from three independent experiments, each N = 3. F ) Colony formation. Colony formation after 14 days was determined in three independent experiments, each N = 2, and expressed as the percent of VC. Values in all graphs are the mean ± SEM. * denotes p≤0.05 between the indicated groups.

    Journal: PLoS ONE

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis

    doi: 10.1371/journal.pone.0102289

    Figure Lengend Snippet: Studies were conducted on MAP2K4 variant cell lines. A ) Knockdown of MAP2K4 protein by siRNA. After transfection of cells with siRNA targeting MAP2K4 (siMAP2K4) or non-targeting control (siCON), MAP2K4 expression was measured by Western blot. Percent knockdown is denoted above the blot. B ) Effect of MAP2K4 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Effects on cell migration. Cell migration was measured as described in . Data from individual types of clones were combined, are from three independent experiments, each in replicates of N = 3, and are expressed as the percentage of migrating cells, normalized to VC. D ) MMP transcript expression. The levels of MMP-2, MMP-9, MMP-10 and MMP-13 transcript levels were measured by qRT/PCR, normalized to GAPDH, and expressed as the percentage of VC. Data are from four independent experiments, each in replicates of N = 2. E ) Cell growth. Cells were plated at the indicated concentrations, allowed to grow for 5 days, MTT added, and optical density determined. Data are from three independent experiments, each N = 3. F ) Colony formation. Colony formation after 14 days was determined in three independent experiments, each N = 2, and expressed as the percent of VC. Values in all graphs are the mean ± SEM. * denotes p≤0.05 between the indicated groups.

    Article Snippet: The following were purchased: antibodies, SEK1/MKK4 (#9152), phospho-p38 MAPK (T180/Y182) (#4631), p38 MAPK (#9212), phospho-SAPK/JNK (T183/Y185) (#4668), SAPK/JNK (#9258), phospho-HSP27 (#2401), HSP27 (#2402), and GAPDH (#2118), all from Cell Signaling Technology, and horseradish peroxidase-conjugated anti-mouse and anti-rabbit, from GE Healthcare Biosciences; plasmids, wild type and constitutive active MAP2K4 (#14615 and #14813; Addgene) and GFP (Vivid Colors pcDNA 6.2/N-EmGFP-GW/TOPO, Invitrogen); chemical inhibitors, p38 MAPK inhibitor, SB203580 (Sigma-Aldrich) and associated negative control, SB202474, and JNK Inhibitor II (SP600125), and JNK Inhibitor II Negative Control (N 1 -Methyl-1,9-pyrazoloanthrone), all from EMD Millipore.

    Techniques: Variant Assay, Knockdown, Transfection, Control, Expressing, Western Blot, Migration, Clone Assay, Quantitative RT-PCR

    A ) MAP2K4 protein expression in LNCaP and 1542CPTX cell lines by Western Blot. B ) Relative cellular invasion of LNCaP and 1542CPTX (42C) cell lines as measured by a Matrigel coated Boyden chamber. Data are from four independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. C ) Relative MMP-2 mRNA expression in LNCaP and 42C cell lines as measured by qRT/PCR. Data are from five independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. D ) Phosphorylated and total HSP27 expression in LNCaP and 1542CPTX cell lines by Western Blot.

    Journal: PLoS ONE

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis

    doi: 10.1371/journal.pone.0102289

    Figure Lengend Snippet: A ) MAP2K4 protein expression in LNCaP and 1542CPTX cell lines by Western Blot. B ) Relative cellular invasion of LNCaP and 1542CPTX (42C) cell lines as measured by a Matrigel coated Boyden chamber. Data are from four independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. C ) Relative MMP-2 mRNA expression in LNCaP and 42C cell lines as measured by qRT/PCR. Data are from five independent experiments, each in replicates of N = 2. * denotes p≤0.05 between the indicated groups. D ) Phosphorylated and total HSP27 expression in LNCaP and 1542CPTX cell lines by Western Blot.

    Article Snippet: The following were purchased: antibodies, SEK1/MKK4 (#9152), phospho-p38 MAPK (T180/Y182) (#4631), p38 MAPK (#9212), phospho-SAPK/JNK (T183/Y185) (#4668), SAPK/JNK (#9258), phospho-HSP27 (#2401), HSP27 (#2402), and GAPDH (#2118), all from Cell Signaling Technology, and horseradish peroxidase-conjugated anti-mouse and anti-rabbit, from GE Healthcare Biosciences; plasmids, wild type and constitutive active MAP2K4 (#14615 and #14813; Addgene) and GFP (Vivid Colors pcDNA 6.2/N-EmGFP-GW/TOPO, Invitrogen); chemical inhibitors, p38 MAPK inhibitor, SB203580 (Sigma-Aldrich) and associated negative control, SB202474, and JNK Inhibitor II (SP600125), and JNK Inhibitor II Negative Control (N 1 -Methyl-1,9-pyrazoloanthrone), all from EMD Millipore.

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    A ) Knockdown of HSP27 protein by siRNA. After transfection of cells with siRNA targeting HSP27 (siHSP27) or non-targeting control (siCON), HSP27 expression was measured by Western blot. B ) Effect of HSP27 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Knockdown of MMP-2 mRNA by siRNA. After transfection of cells with siRNA targeting MMP-2 (siMMP2) or non-targeting control (siCON), MMP-2 expression was measured by qRT/PCR. D ) Effect of MMP-2 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells.

    Journal: PLoS ONE

    Article Title: Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis

    doi: 10.1371/journal.pone.0102289

    Figure Lengend Snippet: A ) Knockdown of HSP27 protein by siRNA. After transfection of cells with siRNA targeting HSP27 (siHSP27) or non-targeting control (siCON), HSP27 expression was measured by Western blot. B ) Effect of HSP27 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells. C ) Knockdown of MMP-2 mRNA by siRNA. After transfection of cells with siRNA targeting MMP-2 (siMMP2) or non-targeting control (siCON), MMP-2 expression was measured by qRT/PCR. D ) Effect of MMP-2 knockdown on cell invasion. MAP2K4 variant cell lines were treated with siRNA, as indicated, and cell invasion measured and depicted as in . Data are from four experiments, each in replicates of N = 2, and are expressed as the percentage of invading cells.

    Article Snippet: The following were purchased: antibodies, SEK1/MKK4 (#9152), phospho-p38 MAPK (T180/Y182) (#4631), p38 MAPK (#9212), phospho-SAPK/JNK (T183/Y185) (#4668), SAPK/JNK (#9258), phospho-HSP27 (#2401), HSP27 (#2402), and GAPDH (#2118), all from Cell Signaling Technology, and horseradish peroxidase-conjugated anti-mouse and anti-rabbit, from GE Healthcare Biosciences; plasmids, wild type and constitutive active MAP2K4 (#14615 and #14813; Addgene) and GFP (Vivid Colors pcDNA 6.2/N-EmGFP-GW/TOPO, Invitrogen); chemical inhibitors, p38 MAPK inhibitor, SB203580 (Sigma-Aldrich) and associated negative control, SB202474, and JNK Inhibitor II (SP600125), and JNK Inhibitor II Negative Control (N 1 -Methyl-1,9-pyrazoloanthrone), all from EMD Millipore.

    Techniques: Knockdown, Transfection, Control, Expressing, Western Blot, Variant Assay, Quantitative RT-PCR