rabbit anti phospho p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho p38 mapk t180 y182
    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the <t>p38</t> pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Rabbit Anti Phospho P38 Mapk T180 Y182, 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 "Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling"

    Article Title: Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling

    Journal: Scientific Reports

    doi: 10.1038/s41598-024-58619-1

    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Figure Legend Snippet: Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).

    Techniques Used: Transformation Assay, Incubation, Sequencing, Western Blot

    MS-based phosphoproteomics of EGF signaling with treatment of PP2C inhibitor sanguinarine in presence of p38 inhibitor BIRB796. ( A ) Schematic illustration of MS-based phosphoproteomics sample preparation. The figure was created using BioRender.com. ( B ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. ( C ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment.
    Figure Legend Snippet: MS-based phosphoproteomics of EGF signaling with treatment of PP2C inhibitor sanguinarine in presence of p38 inhibitor BIRB796. ( A ) Schematic illustration of MS-based phosphoproteomics sample preparation. The figure was created using BioRender.com. ( B ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. ( C ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment.

    Techniques Used: Sample Prep, Transformation Assay

    anti phospho p38 mapk t180 y182 rabbit polyclonal  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 mapk t180 y182 rabbit polyclonal
    Anti Phospho P38 Mapk T180 Y182 Rabbit Polyclonal, 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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    anti rabbit phospho p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti rabbit phospho p38 mapk t180 y182
    Anti Rabbit Phospho P38 Mapk T180 Y182, 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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    rabbit anti phospho p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho p38 mapk t180 y182
    (A) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor (B) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. (C) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. (D) Illustration of the <t>p38</t> pathway activated by EGFR. The figure was created using BioRender.com. (E) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. (F) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also for the heatmap containing all 80 peptides. (G) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Rabbit Anti Phospho P38 Mapk T180 Y182, 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 "Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling"

    Article Title: Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling

    Journal: bioRxiv

    doi: 10.1101/2024.02.01.578427

    (A) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor (B) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. (C) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. (D) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. (E) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. (F) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also for the heatmap containing all 80 peptides. (G) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Figure Legend Snippet: (A) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor (B) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. (C) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. (D) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. (E) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. (F) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also for the heatmap containing all 80 peptides. (G) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).

    Techniques Used: Transformation Assay, Incubation, Sequencing, Western Blot

    anti phospho p38 mapk t180 y182 rabbit polyclonal  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho p38 mapk t180 y182 rabbit polyclonal
    Anti Phospho P38 Mapk T180 Y182 Rabbit Polyclonal, 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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    phospho p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 mapk t180 y182
    VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.
    Phospho P38 Mapk T180 Y182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells"

    Article Title: VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells

    Journal: Cells

    doi: 10.3390/cells12060967

    VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.
    Figure Legend Snippet: VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.

    Techniques Used: Activity Assay, SDS Page, Western Blot, Transduction, Luciferase, Quantitative RT-PCR, Expressing

    Schematic diagram of differential ERK5 activation by VEGF in endothelial cells and EGF in HeLa cells. Ligand-mediated activation of EGFR-1 in HeLa cells activates MEK5 predominantly via MEKK2, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain, resulting in a conformational change in ERK5 and facilitating phosphorylation of ERK5 on multiple residues in the C-terminal transcriptional transactivation domain (TAD). The nuclear localisation sequence (NLS) within the C-terminal domain is now able to promote a nuclear localisation of ERK5. ERK5 is thought to drive MEF2-dependent gene expression by both directly phosphorylating MEF2 transcription factors and by the involvement of the ERK5 TAD. In contrast, ligand-mediated activation of VEGFR-2 in endothelial cells activates MEK5 via MEKK3, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain. However, ERK5 does not undergo phosphorylation in the C-terminus and is unable to translocate to the nucleus, and is instead localised to the cytoplasmic and plasma membrane area ultimately allowing phosphorylation of AKT and suppression of apoptosis in endothelial cells. In contrast to EGFR-1 signalling, MEF2 transcriptional activity in response to VEGFR-2 activation proceeds via p38 MAPK.
    Figure Legend Snippet: Schematic diagram of differential ERK5 activation by VEGF in endothelial cells and EGF in HeLa cells. Ligand-mediated activation of EGFR-1 in HeLa cells activates MEK5 predominantly via MEKK2, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain, resulting in a conformational change in ERK5 and facilitating phosphorylation of ERK5 on multiple residues in the C-terminal transcriptional transactivation domain (TAD). The nuclear localisation sequence (NLS) within the C-terminal domain is now able to promote a nuclear localisation of ERK5. ERK5 is thought to drive MEF2-dependent gene expression by both directly phosphorylating MEF2 transcription factors and by the involvement of the ERK5 TAD. In contrast, ligand-mediated activation of VEGFR-2 in endothelial cells activates MEK5 via MEKK3, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain. However, ERK5 does not undergo phosphorylation in the C-terminus and is unable to translocate to the nucleus, and is instead localised to the cytoplasmic and plasma membrane area ultimately allowing phosphorylation of AKT and suppression of apoptosis in endothelial cells. In contrast to EGFR-1 signalling, MEF2 transcriptional activity in response to VEGFR-2 activation proceeds via p38 MAPK.

    Techniques Used: Activation Assay, Sequencing, Expressing, Activity Assay

    phospho p38 kinase t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho p38 kinase t180 y182
    Phospho P38 Kinase T180 Y182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho p38 kinase t180 y182
    Phospho P38 Kinase T180 Y182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p38 mapk t180 y182
    Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and <t>p38</t> <t>MAPK</t> (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.
    Anti P38 Mapk T180 Y182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Claspin-Dependent and -Independent Chk1 Activation by a Panel of Biological Stresses"

    Article Title: Claspin-Dependent and -Independent Chk1 Activation by a Panel of Biological Stresses

    Journal: Biomolecules

    doi: 10.3390/biom13010125

    Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and p38 MAPK (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.
    Figure Legend Snippet: Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and p38 MAPK (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.

    Techniques Used: Activation Assay, Transfection, Western Blot, Incubation, Concentration Assay

    rabbit anti phospho p38 mapk t180 y182  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit anti phospho p38 mapk t180 y182
    Rabbit Anti Phospho P38 Mapk T180 Y182, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc rabbit anti phospho p38 mapk t180 y182
    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the <t>p38</t> pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Rabbit Anti Phospho P38 Mapk T180 Y182, 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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    Cell Signaling Technology Inc anti phospho p38 mapk t180 y182 rabbit polyclonal
    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the <t>p38</t> pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
    Anti Phospho P38 Mapk T180 Y182 Rabbit Polyclonal, 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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    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the <t>p38</t> pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).
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    VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.
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    VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the <t>p38</t> <t>MAPK</t> inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.
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    Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and <t>p38</t> <t>MAPK</t> (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.
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    Image Search Results


    Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).

    Journal: Scientific Reports

    Article Title: Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling

    doi: 10.1038/s41598-024-58619-1

    Figure Lengend Snippet: Investigation of phosphosites showing EGF-dependent up-regulation. ( A ) Scheme to filter peptides showing up-regulation by inhibitor treatment and summary of the numbers of the up-regulated phosphosites for each inhibitor ( B ) PCA of the log2-transformed abundance of phosphosites up-regulated by both 8 and 20 min of EGF incubation. ( C ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. Proteins with at least one connection with other proteins are included. ( D ) Illustration of the p38 pathway activated by EGFR. The figure was created using BioRender.com. ( E ) Sequence motif enrichment analysis of 15 residues surrounding the regulated phosphosites. ( F ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment. See also Fig. S5 for the heatmap containing all 80 peptides. ( G ) Western blotting analysis using the cell lysates of Hela cells pre-treated with the inhibitors (500 nM) for 15 min followed by 20 min incubation with EGF (20 nM).

    Article Snippet: Primary antibodies used were rabbit anti-EGFR (ab32198; Abcam), rabbit anti-phospho-EGFR (pY1045) (#2237; Cell Signaling Technology), mouse anti-phospho-EGFR (pY1068) (#2236; Cell Signaling Technology), rabbit anti-Erk1/2 (#4695; Cell Signaling Technology), mouse anti-phospho-Erk1/2 (T202/Y204) (#9106; Cell Signaling Technology), rabbit anti-Akt (#9272; Cell Signaling Technology) rabbit anti-phospho-Akt (S473) (#9271; Cell Signaling Technology), rabbit anti-p38 MAPK (#9212; Cell Signaling Technology), rabbit anti-phospho p38 MAPK (T180/Y182) (#9211; Cell Signaling Technology) and mouse anti-GAPDH (Abcam).

    Techniques: Transformation Assay, Incubation, Sequencing, Western Blot

    MS-based phosphoproteomics of EGF signaling with treatment of PP2C inhibitor sanguinarine in presence of p38 inhibitor BIRB796. ( A ) Schematic illustration of MS-based phosphoproteomics sample preparation. The figure was created using BioRender.com. ( B ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. ( C ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment.

    Journal: Scientific Reports

    Article Title: Phosphoproteomic investigation of targets of protein phosphatases in EGFR signaling

    doi: 10.1038/s41598-024-58619-1

    Figure Lengend Snippet: MS-based phosphoproteomics of EGF signaling with treatment of PP2C inhibitor sanguinarine in presence of p38 inhibitor BIRB796. ( A ) Schematic illustration of MS-based phosphoproteomics sample preparation. The figure was created using BioRender.com. ( B ) STRING-based protein network of proteins showing sanguinarine-dependent up-regulation of its phosphorylation. ( C ) Heatmap showing the log2-transformed phosphosite abundance changes induced by inhibitor treatment.

    Article Snippet: Primary antibodies used were rabbit anti-EGFR (ab32198; Abcam), rabbit anti-phospho-EGFR (pY1045) (#2237; Cell Signaling Technology), mouse anti-phospho-EGFR (pY1068) (#2236; Cell Signaling Technology), rabbit anti-Erk1/2 (#4695; Cell Signaling Technology), mouse anti-phospho-Erk1/2 (T202/Y204) (#9106; Cell Signaling Technology), rabbit anti-Akt (#9272; Cell Signaling Technology) rabbit anti-phospho-Akt (S473) (#9271; Cell Signaling Technology), rabbit anti-p38 MAPK (#9212; Cell Signaling Technology), rabbit anti-phospho p38 MAPK (T180/Y182) (#9211; Cell Signaling Technology) and mouse anti-GAPDH (Abcam).

    Techniques: Sample Prep, Transformation Assay

    VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.

    Journal: Cells

    Article Title: VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells

    doi: 10.3390/cells12060967

    Figure Lengend Snippet: VEGF-mediated ERK5 activity is not required for MEF2 transcriptional activity in endothelial cells. ( A ) HDMECs and HeLa cells were serum-starved overnight and then treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 10 min. Cells were lysed in RIPA buffer, and lysates were resolved by 10% SDS-PAGE or Phos-tag SDS-PAGE and analysed by Western blotting. ( B ) HDMECs and HeLa cells were transduced with Ad-MEF2-Luc (MOI 50) and Ad-CMV-LacZ (MOI 20) for 24 h prior to serum starvation overnight. Cells were treated with either vehicle control (0.1% DMSO) or the MEK5 inhibitor BIX02189 (BIX; 1 μM) or the p38 MAPK inhibitor BIRB796 (BIRB; 1 μM), for 1 h prior to stimulation with VEGF (50 ng/mL) or EGF (50 ng/mL) for 6 h. Cells were lysed and MEF2 reporter activity determined by luciferase activity. Relative luciferase activities were normalised to beta galactosidase activity and compared to basal response ± SEM ( n = 3). ( C ) Analysis of MEF2 family mRNA levels by qRT-PCR in HDMECs and HeLa cells. Expression levels of MEF2A , MEF2B , MEF2C, and MEF2D were determined by qRT-PCR and normalised to β-ACTIN level. Results are plotted as fold change relative to level in HeLa cells ± range ( n = 2). Statistical analysis: one-way ANOVA followed by Tukey’s post hoc test, where ** p < 0.01, *** p < 0.001, ns = not significant.

    Article Snippet: Antibodies against total ERK5 (#12950), MEK5 (#91670), p38 MAPK (#8690), Actin (#8456), EGFR1 (#4267) and VEGFR2 (#2479) AKT (#4691), p44/42 MAPK (#4696), MAPKAPK2 (#3042) and antibodies against phospho-AKT S473 (#4060 and Alexa Fluor ® 488 Conjugate (#2336), phospho-p44/42 MAPK T202/Y204 (#4370), phospho-ERK5 T218/Y220 (#3371), phospho-EGFR1 Y1061 (#3777), phospho-VEGFR2 Y1175 (#3770), phospho-p38 MAPK T180/Y182 (#4631), phospho-MAPKAPK2 Thr334 (#3007) were purchased from Cell Signalling Technology (Leiden, Netherlands).

    Techniques: Activity Assay, SDS Page, Western Blot, Transduction, Luciferase, Quantitative RT-PCR, Expressing

    Schematic diagram of differential ERK5 activation by VEGF in endothelial cells and EGF in HeLa cells. Ligand-mediated activation of EGFR-1 in HeLa cells activates MEK5 predominantly via MEKK2, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain, resulting in a conformational change in ERK5 and facilitating phosphorylation of ERK5 on multiple residues in the C-terminal transcriptional transactivation domain (TAD). The nuclear localisation sequence (NLS) within the C-terminal domain is now able to promote a nuclear localisation of ERK5. ERK5 is thought to drive MEF2-dependent gene expression by both directly phosphorylating MEF2 transcription factors and by the involvement of the ERK5 TAD. In contrast, ligand-mediated activation of VEGFR-2 in endothelial cells activates MEK5 via MEKK3, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain. However, ERK5 does not undergo phosphorylation in the C-terminus and is unable to translocate to the nucleus, and is instead localised to the cytoplasmic and plasma membrane area ultimately allowing phosphorylation of AKT and suppression of apoptosis in endothelial cells. In contrast to EGFR-1 signalling, MEF2 transcriptional activity in response to VEGFR-2 activation proceeds via p38 MAPK.

    Journal: Cells

    Article Title: VEGF Stimulates Activation of ERK5 in the Absence of C-Terminal Phosphorylation Preventing Nuclear Localization and Facilitating AKT Activation in Endothelial Cells

    doi: 10.3390/cells12060967

    Figure Lengend Snippet: Schematic diagram of differential ERK5 activation by VEGF in endothelial cells and EGF in HeLa cells. Ligand-mediated activation of EGFR-1 in HeLa cells activates MEK5 predominantly via MEKK2, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain, resulting in a conformational change in ERK5 and facilitating phosphorylation of ERK5 on multiple residues in the C-terminal transcriptional transactivation domain (TAD). The nuclear localisation sequence (NLS) within the C-terminal domain is now able to promote a nuclear localisation of ERK5. ERK5 is thought to drive MEF2-dependent gene expression by both directly phosphorylating MEF2 transcription factors and by the involvement of the ERK5 TAD. In contrast, ligand-mediated activation of VEGFR-2 in endothelial cells activates MEK5 via MEKK3, resulting in the phosphorylation of ERK5 on the activation-loop T-E-Y motif in the ERK5 kinase domain. However, ERK5 does not undergo phosphorylation in the C-terminus and is unable to translocate to the nucleus, and is instead localised to the cytoplasmic and plasma membrane area ultimately allowing phosphorylation of AKT and suppression of apoptosis in endothelial cells. In contrast to EGFR-1 signalling, MEF2 transcriptional activity in response to VEGFR-2 activation proceeds via p38 MAPK.

    Article Snippet: Antibodies against total ERK5 (#12950), MEK5 (#91670), p38 MAPK (#8690), Actin (#8456), EGFR1 (#4267) and VEGFR2 (#2479) AKT (#4691), p44/42 MAPK (#4696), MAPKAPK2 (#3042) and antibodies against phospho-AKT S473 (#4060 and Alexa Fluor ® 488 Conjugate (#2336), phospho-p44/42 MAPK T202/Y204 (#4370), phospho-ERK5 T218/Y220 (#3371), phospho-EGFR1 Y1061 (#3777), phospho-VEGFR2 Y1175 (#3770), phospho-p38 MAPK T180/Y182 (#4631), phospho-MAPKAPK2 Thr334 (#3007) were purchased from Cell Signalling Technology (Leiden, Netherlands).

    Techniques: Activation Assay, Sequencing, Expressing, Activity Assay

    Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and p38 MAPK (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.

    Journal: Biomolecules

    Article Title: Claspin-Dependent and -Independent Chk1 Activation by a Panel of Biological Stresses

    doi: 10.3390/biom13010125

    Figure Lengend Snippet: Effects of Claspin or ATR depletion on Chk1 activation by various biological stresses and on the factors involved in growth-related pathways. ( A ) Claspin(f/-) MEF cells were treated with Ad-Cre or nontreated and exposed to various stresses for 3 h. ( B , D ) HeLa cells were transfected with siRNA for Claspin ( B ) or for ATR ( C ), and were exposed to indicated stresses for 3 h before the harvest. The whole cell extracts were analyzed by Western blotting with antibodies indicated. SAPK/JUNK and p38 MAPK (MAPK activated by stresses and growth factors); ERK1/2 (MAPK activated by growth factors and mitogen); 4EBP1 (phosphorylated by mTOR and required for activation of translation); PDK1 (required for activation of mTOR); Mcm2 (phosphorylated by Cdc7 and required for replication); Mcl-1 (anti-apoptotic factor). Phosphorylated forms represent activated states. -, control siRNA. ( D ) Extracts prepared from HeLa cells treated with the stresses indicated were incubated in the absence of presence of λppase, and analyzed on a low concentration gel.

    Article Snippet: Anti-Chk1 phospho-S345 (#2348), anti-Chk1 phospho-S317 (#2344), anti-p44/42 MAPK (Erk1/2) (#4695), anti- SAPK/JNK (#9252), p38 MAPK (#8690), anti-p38 MAPK T180/Y182 (#4511), anti-p44/42 MAPK (Erk1/2) T202/Y204 (#4370), anti-SAPK/JNK T183/Y185 (#4668), Caspase-9 (#9508), Cleaved Caspase-3(#9661), and Mcl-1 (#5453) were obtained from Cell Signaling.

    Techniques: Activation Assay, Transfection, Western Blot, Incubation, Concentration Assay