sb203580  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Dimethyl
    Description:

    Catalog Number:
    901460
    Price:
    None
    Buy from Supplier


    Structured Review

    Millipore sb203580
    Dimethyl

    https://www.bioz.com/result/sb203580/product/Millipore
    Average 99 stars, based on 1013 article reviews
    Price from $9.99 to $1999.99
    sb203580 - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Ursolic acid inhibits cell migration and promotes JNK-dependent lysosomal associated cell death in Glioblastoma multiforme cells"

    Article Title: Ursolic acid inhibits cell migration and promotes JNK-dependent lysosomal associated cell death in Glioblastoma multiforme cells

    Journal: bioRxiv

    doi: 10.1101/2020.03.11.987578

    UA induces Caspase-independent, JNK-dependent cell death. ( A ) Cells were pre-loaded 12.5µM SP600125 JNK inhibitior for 1 hr, increasing concentrations of UA were added in the presence of 12.5µM SP600125 and incubated for 48hrs. Cells were then analysed using Alamar blue cell viability assay. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni post test. ( B ) U373MG cells were pre-treated with 50µM zVAD-FMK for 1hr prior to UA treatment. Cells were then incubated for 48hrs and analysed by Alamar blue. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni’s post test. ( C ) Cells were pre-loaded 10µM SB203580 p38-MAPK inihibitor for 1 hr, increasing concentrations of UA were added and incubated for 48 hrs. Cells were then analylsed using Alamar blue cell viability assay. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni post test ( D ) Cells were pre-treated with either zVAD-fmk (50μM) or SP600125 (12.5μM) for 1 hrprior to addition of TMZ (26.5μM) for 6 days. Cell viability was analysed by Alamar blue. Statistical analysis was carried out using One-Way ANOVA with Tukey’s post-test (p
    Figure Legend Snippet: UA induces Caspase-independent, JNK-dependent cell death. ( A ) Cells were pre-loaded 12.5µM SP600125 JNK inhibitior for 1 hr, increasing concentrations of UA were added in the presence of 12.5µM SP600125 and incubated for 48hrs. Cells were then analysed using Alamar blue cell viability assay. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni post test. ( B ) U373MG cells were pre-treated with 50µM zVAD-FMK for 1hr prior to UA treatment. Cells were then incubated for 48hrs and analysed by Alamar blue. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni’s post test. ( C ) Cells were pre-loaded 10µM SB203580 p38-MAPK inihibitor for 1 hr, increasing concentrations of UA were added and incubated for 48 hrs. Cells were then analylsed using Alamar blue cell viability assay. Statistical analysis was carried out using Two-Way ANOVA with Bonferroni post test ( D ) Cells were pre-treated with either zVAD-fmk (50μM) or SP600125 (12.5μM) for 1 hrprior to addition of TMZ (26.5μM) for 6 days. Cell viability was analysed by Alamar blue. Statistical analysis was carried out using One-Way ANOVA with Tukey’s post-test (p

    Techniques Used: Incubation, Viability Assay

    2) Product Images from "Involvement of Mitogen-Activated Protein Kinase Pathways in Expression of the Water Channel Protein Aquaporin-4 after Ischemia in Rat Cortical Astrocytes"

    Article Title: Involvement of Mitogen-Activated Protein Kinase Pathways in Expression of the Water Channel Protein Aquaporin-4 after Ischemia in Rat Cortical Astrocytes

    Journal: Journal of Neurotrauma

    doi: 10.1089/neu.2012.2430

    LDH release after OGD injury in rat cortical astrocytes. Cells were subjected to 6 h of OGD, followed by 24 h of reoxygenation. Cell death was assessed by LDH release. SB203580 (10 μM) significantly reduced OGD-induced
    Figure Legend Snippet: LDH release after OGD injury in rat cortical astrocytes. Cells were subjected to 6 h of OGD, followed by 24 h of reoxygenation. Cell death was assessed by LDH release. SB203580 (10 μM) significantly reduced OGD-induced

    Techniques Used:

    Infarct volume and edema volume 3 days after transient focal cerebral ischemia in vehicle-treated and SB203580-treated rats ( n =6 each; * p
    Figure Legend Snippet: Infarct volume and edema volume 3 days after transient focal cerebral ischemia in vehicle-treated and SB203580-treated rats ( n =6 each; * p

    Techniques Used:

    The cells were treated with 10 μM or 1 μM of PD98059, SP600125, or SB203580, before OGD. Sixteen hours after reoxygenation, the AQP4 level was determined by Western blot analysis. Ten micromoles of SB203580 or SP600125
    Figure Legend Snippet: The cells were treated with 10 μM or 1 μM of PD98059, SP600125, or SB203580, before OGD. Sixteen hours after reoxygenation, the AQP4 level was determined by Western blot analysis. Ten micromoles of SB203580 or SP600125

    Techniques Used: Western Blot

    ( A ) Suppression of p38 MAPK activity by administration of SB203580. p38 MAPK activity was examined 1 and 3 days after reperfusion with or without SB203580 administration. SB203580 was administered 30 min before ischemic insult, and phosphorylation
    Figure Legend Snippet: ( A ) Suppression of p38 MAPK activity by administration of SB203580. p38 MAPK activity was examined 1 and 3 days after reperfusion with or without SB203580 administration. SB203580 was administered 30 min before ischemic insult, and phosphorylation

    Techniques Used: Activity Assay

    3) Product Images from "Synergistic Induction of Cyclooxygenase-2 by Transforming Growth Factor-β1 and Epidermal Growth Factor Inhibits Apoptosis in Epithelial Cells 1"

    Article Title: Synergistic Induction of Cyclooxygenase-2 by Transforming Growth Factor-β1 and Epidermal Growth Factor Inhibits Apoptosis in Epithelial Cells 1

    Journal: Neoplasia (New York, N.Y.)

    doi:

    TGF-β1- and EGF-mediated induction of COX-2 is inhibited by PD98059 (75 µM), SB203580 (10 µM) and AG1478 (50 µM) in Mv1Lu cells. PD98059, SB203580 and AG1478 compounds were added in the serum-free media 1 hour prior to the addition of TGF-β1 and EGF. After 8 hours of treatment with the growth factors, cells were lysed and 50 µg of each cell lysate was used for immunoblotting.
    Figure Legend Snippet: TGF-β1- and EGF-mediated induction of COX-2 is inhibited by PD98059 (75 µM), SB203580 (10 µM) and AG1478 (50 µM) in Mv1Lu cells. PD98059, SB203580 and AG1478 compounds were added in the serum-free media 1 hour prior to the addition of TGF-β1 and EGF. After 8 hours of treatment with the growth factors, cells were lysed and 50 µg of each cell lysate was used for immunoblotting.

    Techniques Used:

    4) Product Images from "Ras and Rap signal bidirectional synaptic plasticity via distinct subcellular microdomains"

    Article Title: Ras and Rap signal bidirectional synaptic plasticity via distinct subcellular microdomains

    Journal: Neuron

    doi: 10.1016/j.neuron.2018.03.049

    Rap signals JNK and p38MAPK in the bulk membrane and lysosome (A) Schematic drawing outlines in vitro experimental design. (B) Evoked AMPA-R- (−60 mV) and NMDA-R- (+40 mV) mediated responses recorded from control non-expressing (Ctrl), CD8-Rap2(ca)-IRES-GFP and LAMP1-Rap1(ca)-IRES-RFP expressing CA1 cells cultured in media containing SP600125 or SB203580. (C–D) AMPA and NMDA responses in CD8-Rap2(ca) and LAMP1-Rap1(ca) expressing neurons relative to non-expressing CA1 cells cultured in media containing 5 μM SP600125 ( C ) or 2 μM SB203580 ( D for values. (E–F) AMPA and NMDA responses in CD8-Rap2(dn) and LAMP1-Rap1(dn) expressing neurons relative to non-expressing CA1 cells cultured in media containing 5 μM SP600125 ( E ) or 2 μM SB203580 ( F for values. (G) Evoked AMPA and NMDA responses recorded from non-expressing (Ctrl), CD8-Rap2(ca)-IRES-GFP and LAMP1-Rap1(ca)-IRES-RFP expressing CA1 cells in cultured slices prepared from GluA2 knockout and GluA2(K882A) transgenic mice. (H–J) AMPA and NMDA responses in CD8-Rap2(ca) and LAMP1-Rap1(ca) expressing neurons relative to non-expressing CA1 cells in cultured slices prepared from GluA1 knockout ( H ), GluA2 knockout ( I ) and GluA2(K882A) transgenic ( J for values. (K–L) AMPA and NMDA responses in CD8-Rap2(dn) and LAMP1-Rap1(dn) expressing neurons relative to non-expressing CA1 cells in cultured slices prepared from GluA1 knockout ( K ) and GluA2 knockout ( L for values. Asterisks indicate p
    Figure Legend Snippet: Rap signals JNK and p38MAPK in the bulk membrane and lysosome (A) Schematic drawing outlines in vitro experimental design. (B) Evoked AMPA-R- (−60 mV) and NMDA-R- (+40 mV) mediated responses recorded from control non-expressing (Ctrl), CD8-Rap2(ca)-IRES-GFP and LAMP1-Rap1(ca)-IRES-RFP expressing CA1 cells cultured in media containing SP600125 or SB203580. (C–D) AMPA and NMDA responses in CD8-Rap2(ca) and LAMP1-Rap1(ca) expressing neurons relative to non-expressing CA1 cells cultured in media containing 5 μM SP600125 ( C ) or 2 μM SB203580 ( D for values. (E–F) AMPA and NMDA responses in CD8-Rap2(dn) and LAMP1-Rap1(dn) expressing neurons relative to non-expressing CA1 cells cultured in media containing 5 μM SP600125 ( E ) or 2 μM SB203580 ( F for values. (G) Evoked AMPA and NMDA responses recorded from non-expressing (Ctrl), CD8-Rap2(ca)-IRES-GFP and LAMP1-Rap1(ca)-IRES-RFP expressing CA1 cells in cultured slices prepared from GluA2 knockout and GluA2(K882A) transgenic mice. (H–J) AMPA and NMDA responses in CD8-Rap2(ca) and LAMP1-Rap1(ca) expressing neurons relative to non-expressing CA1 cells in cultured slices prepared from GluA1 knockout ( H ), GluA2 knockout ( I ) and GluA2(K882A) transgenic ( J for values. (K–L) AMPA and NMDA responses in CD8-Rap2(dn) and LAMP1-Rap1(dn) expressing neurons relative to non-expressing CA1 cells in cultured slices prepared from GluA1 knockout ( K ) and GluA2 knockout ( L for values. Asterisks indicate p

    Techniques Used: In Vitro, Expressing, Cell Culture, Knock-Out, Transgenic Assay, Mouse Assay

    5) Product Images from "Activin A Stimulates Mouse APCs to Express BAFF via ALK4-Smad3 Pathway"

    Article Title: Activin A Stimulates Mouse APCs to Express BAFF via ALK4-Smad3 Pathway

    Journal: Immune Network

    doi: 10.4110/in.2011.11.4.196

    Activin A enhances BAFF expression through ALK4 and Smad3. (A) Activin A induces BAFF mRNA expression via ALK4. DC2.4 cells were pre-treated with 10µM SB431542, 10µM SB203580, 10µM SP600125, or 5µM PD98059 for 1 h and then stimulated with activin A (10 ng/ml) for 24 h. Levels of BAFF transcripts were determined by RT-PCR. (B) DC2.4 cells (1×10 6 ) were transfected with DN-Smad3 expression vectors (2, 4, 8µg) and stimulated with activin A (10 ng/ml) for 24 h. Levels of BAFF transcripts were determined by RT-PCR. (C) BAFF secretion by DN-Smad3-transfected DC 2.4 was measured by ELISA after 48 h incubation with activin A. Data are means of triplicate samples±SEM. ** p
    Figure Legend Snippet: Activin A enhances BAFF expression through ALK4 and Smad3. (A) Activin A induces BAFF mRNA expression via ALK4. DC2.4 cells were pre-treated with 10µM SB431542, 10µM SB203580, 10µM SP600125, or 5µM PD98059 for 1 h and then stimulated with activin A (10 ng/ml) for 24 h. Levels of BAFF transcripts were determined by RT-PCR. (B) DC2.4 cells (1×10 6 ) were transfected with DN-Smad3 expression vectors (2, 4, 8µg) and stimulated with activin A (10 ng/ml) for 24 h. Levels of BAFF transcripts were determined by RT-PCR. (C) BAFF secretion by DN-Smad3-transfected DC 2.4 was measured by ELISA after 48 h incubation with activin A. Data are means of triplicate samples±SEM. ** p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Enzyme-linked Immunosorbent Assay, Incubation

    6) Product Images from "Biphasic Regulation of p38 MAPK by Serotonin Contributes to the Efficacy of Stimulus Protocols That Induce Long-Term Synaptic Facilitation"

    Article Title: Biphasic Regulation of p38 MAPK by Serotonin Contributes to the Efficacy of Stimulus Protocols That Induce Long-Term Synaptic Facilitation

    Journal: eNeuro

    doi: 10.1523/ENEURO.0373-16.2017

    The effect of p38 MAPK inhibitor on pERK. A , Representative confocal images of pERK immunofluorescence in SNs at 60 min after onset of a 5-min pulse of 5-HT, in the absence or presence of the p38 MAPK inhibitor SB203580 (SB). B , Summary data. Compared with the 5-HT alone group, treatment with 5-HT in the presence of SB induced a significant increase in pERK at 60 min after onset of 5-HT. Scale bar, 20 μm. Significant differences are indicated by * for p
    Figure Legend Snippet: The effect of p38 MAPK inhibitor on pERK. A , Representative confocal images of pERK immunofluorescence in SNs at 60 min after onset of a 5-min pulse of 5-HT, in the absence or presence of the p38 MAPK inhibitor SB203580 (SB). B , Summary data. Compared with the 5-HT alone group, treatment with 5-HT in the presence of SB induced a significant increase in pERK at 60 min after onset of 5-HT. Scale bar, 20 μm. Significant differences are indicated by * for p

    Techniques Used: Immunofluorescence

    7) Product Images from "Biphasic Regulation of p38 MAPK by Serotonin Contributes to the Efficacy of Stimulus Protocols That Induce Long-Term Synaptic Facilitation"

    Article Title: Biphasic Regulation of p38 MAPK by Serotonin Contributes to the Efficacy of Stimulus Protocols That Induce Long-Term Synaptic Facilitation

    Journal: eNeuro

    doi: 10.1523/ENEURO.0373-16.2017

    The effect of p38 MAPK inhibitor on pERK. A , Representative confocal images of pERK immunofluorescence in SNs at 60 min after onset of a 5-min pulse of 5-HT, in the absence or presence of the p38 MAPK inhibitor SB203580 (SB). B , Summary data. Compared with the 5-HT alone group, treatment with 5-HT in the presence of SB induced a significant increase in pERK at 60 min after onset of 5-HT. Scale bar, 20 μm. Significant differences are indicated by * for p
    Figure Legend Snippet: The effect of p38 MAPK inhibitor on pERK. A , Representative confocal images of pERK immunofluorescence in SNs at 60 min after onset of a 5-min pulse of 5-HT, in the absence or presence of the p38 MAPK inhibitor SB203580 (SB). B , Summary data. Compared with the 5-HT alone group, treatment with 5-HT in the presence of SB induced a significant increase in pERK at 60 min after onset of 5-HT. Scale bar, 20 μm. Significant differences are indicated by * for p

    Techniques Used: Immunofluorescence

    8) Product Images from "Integrin ?2?1 Promotes Activation of Protein Phosphatase 2A and Dephosphorylation of Akt and Glycogen Synthase Kinase 3?"

    Article Title: Integrin ?2?1 Promotes Activation of Protein Phosphatase 2A and Dephosphorylation of Akt and Glycogen Synthase Kinase 3?

    Journal: Molecular and Cellular Biology

    doi:

    Activation of PP2A by α2β1 integrin is dependent on Cdc42. (A) The effect of the transfected dominant-negative mutant forms of Cdc42 (Cdc42DN), Rac1 (Rac1DN), and RhoA (RhoADN) on the activation of PP2A in Saos-α2 cells. Phosphatase activity relative to cell lysate protein content is shown. Immunoblotting with antibodies against PP2A and Cdc42 was performed to show that Cdc42 was overexpressed in transfected cells and that it had no effect on the amount of PP2A protein in cells. (B) Serum-starved fibroblasts were detached and left untreated or treated with inhibitor BIM I (10 μM) or SB203580 (20 μM) and placed inside collagen for 1 h. Phosphatase activity relative to cell lysate protein content is shown (mean ± SEM). BIM I, four separate experiments; SB203580, two separate experiments ( n = 3 for each).
    Figure Legend Snippet: Activation of PP2A by α2β1 integrin is dependent on Cdc42. (A) The effect of the transfected dominant-negative mutant forms of Cdc42 (Cdc42DN), Rac1 (Rac1DN), and RhoA (RhoADN) on the activation of PP2A in Saos-α2 cells. Phosphatase activity relative to cell lysate protein content is shown. Immunoblotting with antibodies against PP2A and Cdc42 was performed to show that Cdc42 was overexpressed in transfected cells and that it had no effect on the amount of PP2A protein in cells. (B) Serum-starved fibroblasts were detached and left untreated or treated with inhibitor BIM I (10 μM) or SB203580 (20 μM) and placed inside collagen for 1 h. Phosphatase activity relative to cell lysate protein content is shown (mean ± SEM). BIM I, four separate experiments; SB203580, two separate experiments ( n = 3 for each).

    Techniques Used: Activation Assay, Transfection, Dominant Negative Mutation, Activity Assay

    9) Product Images from "Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media "

    Article Title: Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4662-4667.2004

    Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P
    Figure Legend Snippet: Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P

    Techniques Used: Cell Culture, Inhibition

    Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.
    Figure Legend Snippet: Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.

    Techniques Used: Cell Culture

    10) Product Images from "Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media "

    Article Title: Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4662-4667.2004

    Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P
    Figure Legend Snippet: Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P

    Techniques Used: Cell Culture, Inhibition

    Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.
    Figure Legend Snippet: Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.

    Techniques Used: Cell Culture

    11) Product Images from "Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media "

    Article Title: Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4662-4667.2004

    Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P
    Figure Legend Snippet: Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P

    Techniques Used: Cell Culture, Inhibition

    Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.
    Figure Legend Snippet: Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.

    Techniques Used: Cell Culture

    12) Product Images from "Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media "

    Article Title: Role of p38 Mitogen-Activated Protein Kinase in Middle Ear Mucosa Hyperplasia during Bacterial Otitis Media

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4662-4667.2004

    Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P
    Figure Legend Snippet: Average surface area (in square millimeters) of bacterially exposed middle ear mucosal explants (harvested 48 h after bacterial inoculation of the middle ear) cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth each day. Significant inhibition of growth was observed at 10 nM ( P

    Techniques Used: Cell Culture, Inhibition

    Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.
    Figure Legend Snippet: Average surface area (in square millimeters) of noninfected middle ear mucosal explants cultured with different nanomolar concentrations of SB203580 for 10 days and measured for area of growth on days 1, 3, 7, and 10. No effect of the inhibitor on explant growth was observed. Bars represent means, and vertical lines represent 1 SEM.

    Techniques Used: Cell Culture

    13) Product Images from "Atorvastatin attenuates homocysteine-induced apoptosis in human umbilical vein endothelial cells via inhibiting NADPH oxidase-related oxidative stress-triggered p38MAPK signaling"

    Article Title: Atorvastatin attenuates homocysteine-induced apoptosis in human umbilical vein endothelial cells via inhibiting NADPH oxidase-related oxidative stress-triggered p38MAPK signaling

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2009.135

    Atorvastatin inhibited HUVECs apoptosis induced by Hcy involving a MAPK mechanism. (A) Cells were pre-incubated with the p38 inhibitor, SB203580 (10 μmol/L) for 30 min before the addition of 1 mmol/L Hcy for 24 h and apoptosis cells were counted. (B) Cells were stimulated with different concentrations of Hcy (0, 0.1, 0.5, 1.0, 5.0, and 10 mmol/L) for 30 min. (C) Cells were stimulated with 1 mmol/L Hcy for 0, 15, 30, 45, 60, and 90 min, respectively. (D) Effects of atorvastatin, DPI, NAC, and SB203580 on the phosphorylation of p38 MAPK induced by Hcy. After treatment with atorvastatin (10 μmol/L), DPI (10 μmol/L), NAC (1 mmol/L), and 203580 (10 μmol/L) for 30 min, the cells were stimulated by Hcy (1 mmol/L) for 30 min or not. The blots show representative immunoblots of the phosphorylation of p38 MAPK and the graphs show the quantification of the bands by densitometry. Date are normalized with β-actin so that value of the control group is regarded as 1.0. Values are expressed as means±SD from three separate experiments. b P
    Figure Legend Snippet: Atorvastatin inhibited HUVECs apoptosis induced by Hcy involving a MAPK mechanism. (A) Cells were pre-incubated with the p38 inhibitor, SB203580 (10 μmol/L) for 30 min before the addition of 1 mmol/L Hcy for 24 h and apoptosis cells were counted. (B) Cells were stimulated with different concentrations of Hcy (0, 0.1, 0.5, 1.0, 5.0, and 10 mmol/L) for 30 min. (C) Cells were stimulated with 1 mmol/L Hcy for 0, 15, 30, 45, 60, and 90 min, respectively. (D) Effects of atorvastatin, DPI, NAC, and SB203580 on the phosphorylation of p38 MAPK induced by Hcy. After treatment with atorvastatin (10 μmol/L), DPI (10 μmol/L), NAC (1 mmol/L), and 203580 (10 μmol/L) for 30 min, the cells were stimulated by Hcy (1 mmol/L) for 30 min or not. The blots show representative immunoblots of the phosphorylation of p38 MAPK and the graphs show the quantification of the bands by densitometry. Date are normalized with β-actin so that value of the control group is regarded as 1.0. Values are expressed as means±SD from three separate experiments. b P

    Techniques Used: Incubation, Western Blot

    14) Product Images from "Role of MAPK in apolipoprotein CIII-induced apoptosis in INS-1E cells"

    Article Title: Role of MAPK in apolipoprotein CIII-induced apoptosis in INS-1E cells

    Journal: Lipids in Health and Disease

    doi: 10.1186/1476-511X-8-3

    ApoCIII-induced apoptosis . INS-1E cells were cultured for 24 hours in the absence or presence of 10 μg/ml apoCIII. Cells were exposed to 10 μM of the p38 inhibitor SB203580 (SB) or 100 μM of the ERK1/2 inhibitor PD98059 (PD) during 0.5 hours prior to apoCIII treatment as indicated. After culture, apoptosis was measured as DNA fragmentation and normalized to DNA content. *P
    Figure Legend Snippet: ApoCIII-induced apoptosis . INS-1E cells were cultured for 24 hours in the absence or presence of 10 μg/ml apoCIII. Cells were exposed to 10 μM of the p38 inhibitor SB203580 (SB) or 100 μM of the ERK1/2 inhibitor PD98059 (PD) during 0.5 hours prior to apoCIII treatment as indicated. After culture, apoptosis was measured as DNA fragmentation and normalized to DNA content. *P

    Techniques Used: Cell Culture

    15) Product Images from "Bone-derived SDF-1 stimulates IL-6 release via CXCR4, ERK and NF-?B pathways and promotes osteoclastogenesis in human oral cancer cells"

    Article Title: Bone-derived SDF-1 stimulates IL-6 release via CXCR4, ERK and NF-?B pathways and promotes osteoclastogenesis in human oral cancer cells

    Journal: Carcinogenesis

    doi: 10.1093/carcin/bgn045

    ERK is involved in the potentiation of IL-6 expression by SDF-1α. SCC4 cells were incubated with SDF-1α (100 ng/ml) for indicated time intervals, and p-ERK, p-p38, p-JNK or p-Akt expression was determined by western blot analysis ( A ). Cell were pretreated for 30 min with PD98059 (10 μM), SB203580 (10 μM), SP600125 (10 μM) and Akt inhibitor (10 μM) ( B ) or transfected with dominant-negative (DN) mutant of ERK, p38, JNK and Akt ( C ) for 24 h followed by stimulation with SDF-1α (100 ng/ml) for 24 h. Media were collected to measure IL-6. Results are expressed as the mean ± SE. * P
    Figure Legend Snippet: ERK is involved in the potentiation of IL-6 expression by SDF-1α. SCC4 cells were incubated with SDF-1α (100 ng/ml) for indicated time intervals, and p-ERK, p-p38, p-JNK or p-Akt expression was determined by western blot analysis ( A ). Cell were pretreated for 30 min with PD98059 (10 μM), SB203580 (10 μM), SP600125 (10 μM) and Akt inhibitor (10 μM) ( B ) or transfected with dominant-negative (DN) mutant of ERK, p38, JNK and Akt ( C ) for 24 h followed by stimulation with SDF-1α (100 ng/ml) for 24 h. Media were collected to measure IL-6. Results are expressed as the mean ± SE. * P

    Techniques Used: Expressing, Incubation, Western Blot, Transfection, Dominant Negative Mutation, Mutagenesis

    16) Product Images from "Activation of p38 MAPK in CD4 T cells controls IL-17 production and autoimmune encephalomyelitis"

    Article Title: Activation of p38 MAPK in CD4 T cells controls IL-17 production and autoimmune encephalomyelitis

    Journal: Blood

    doi: 10.1182/blood-2011-02-336552

    p38 MAPK controls IL-17 production at the posttranscriptional level. (A) FACS-sorted CD4 T cells from WT B6 mice were differentiated into Th17 cells in the absence or presence of SB203580 (5μM) for the indicated periods of time. The phosphorylation of STAT3 at Ser727 (P-Ser) or at Tyr705 (P-Tyr) was examined by Western blot analysis. Total STAT3 (Total) is also shown. FACS-sorted CD4 T cells from WT mice were activated under Th17 conditions in the absence (control) or presence of SB203580 for 48h. Relative Rorc (B) and Il17 (C) mRNA levels were examined by quantitative real-time PCR using β2-microglobulin as the endogenous control. (D) Relative Il17 mRNA levels in FACS-sorted CD4 T cells from WT B6 and MKK3 −/− MKK6 +/− mice activated under Th17 conditions for 48h. (E) Intracellular staining for IL-17 and IFNγ in WT total CD4 T cells differentiated into Th17 cells in the absence (control) or presence of SB203580 for 72 hours.
    Figure Legend Snippet: p38 MAPK controls IL-17 production at the posttranscriptional level. (A) FACS-sorted CD4 T cells from WT B6 mice were differentiated into Th17 cells in the absence or presence of SB203580 (5μM) for the indicated periods of time. The phosphorylation of STAT3 at Ser727 (P-Ser) or at Tyr705 (P-Tyr) was examined by Western blot analysis. Total STAT3 (Total) is also shown. FACS-sorted CD4 T cells from WT mice were activated under Th17 conditions in the absence (control) or presence of SB203580 for 48h. Relative Rorc (B) and Il17 (C) mRNA levels were examined by quantitative real-time PCR using β2-microglobulin as the endogenous control. (D) Relative Il17 mRNA levels in FACS-sorted CD4 T cells from WT B6 and MKK3 −/− MKK6 +/− mice activated under Th17 conditions for 48h. (E) Intracellular staining for IL-17 and IFNγ in WT total CD4 T cells differentiated into Th17 cells in the absence (control) or presence of SB203580 for 72 hours.

    Techniques Used: FACS, Mouse Assay, Western Blot, Real-time Polymerase Chain Reaction, Staining

    p38 MAPK controls IL-17 production by Th17 cells in vivo. (A) Mononuclear cells were isolated from the CNS of 2× MOG 35-55 -CFA immunized mice on day 35 after immunization, stimulated with PMA/ionomycin for 4 hours in the presence of brefeldin A, stained, and analyzed by flow cytometry. (Left panel) Cells were gated on CD45 and the frequency of CD4 + TCRβ + cells was determined. (Right panels) Cells were gated on CD4 and TCRβ and the frequency of IL-17 + and IFNγ + cells was determined. (B) Spleen and draining lymph node (DLN) cells from 2× MOG 35-55 -CFA immunized mice treated with either carrier (n = 7) or the SB203580 (n = 8) were harvested on day 21 after immunization and stimulated with MOG 35-55 (50 μg/mL) for 72 hours and IL-17 and IFNγ in the supernatants was quantified by ELISA. (C) Spleen and DLN cells from 2× MOG 35-55 -CFA immunized mice (n = 10) were harvested on day 10 after immunization and stimulated with MOG 35-55 in the presence or absence of SB203580 (SB; 5μM) for 72 hours. IL-17 production was quantified by ELISA. (D) Cells obtained as in panel C were stimulated with the indicated amounts of MOG 35-55 in the presence or the absence of SB203580 and proliferation was assessed by [ 3 H]-thymidine incorporation. The significance of differences observed in panels A through C was determined using the Student t test (* ≤ 0.05; *** ≤ 0.001), and by 2-way ANOVA for panel D (SB, P = .31; MOG 35-55 , P = .003; and interaction, P = .95).
    Figure Legend Snippet: p38 MAPK controls IL-17 production by Th17 cells in vivo. (A) Mononuclear cells were isolated from the CNS of 2× MOG 35-55 -CFA immunized mice on day 35 after immunization, stimulated with PMA/ionomycin for 4 hours in the presence of brefeldin A, stained, and analyzed by flow cytometry. (Left panel) Cells were gated on CD45 and the frequency of CD4 + TCRβ + cells was determined. (Right panels) Cells were gated on CD4 and TCRβ and the frequency of IL-17 + and IFNγ + cells was determined. (B) Spleen and draining lymph node (DLN) cells from 2× MOG 35-55 -CFA immunized mice treated with either carrier (n = 7) or the SB203580 (n = 8) were harvested on day 21 after immunization and stimulated with MOG 35-55 (50 μg/mL) for 72 hours and IL-17 and IFNγ in the supernatants was quantified by ELISA. (C) Spleen and DLN cells from 2× MOG 35-55 -CFA immunized mice (n = 10) were harvested on day 10 after immunization and stimulated with MOG 35-55 in the presence or absence of SB203580 (SB; 5μM) for 72 hours. IL-17 production was quantified by ELISA. (D) Cells obtained as in panel C were stimulated with the indicated amounts of MOG 35-55 in the presence or the absence of SB203580 and proliferation was assessed by [ 3 H]-thymidine incorporation. The significance of differences observed in panels A through C was determined using the Student t test (* ≤ 0.05; *** ≤ 0.001), and by 2-way ANOVA for panel D (SB, P = .31; MOG 35-55 , P = .003; and interaction, P = .95).

    Techniques Used: In Vivo, Isolation, Mouse Assay, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

    p38 MAPK regulates IL-17 production by in vitro–generated Th17 cells. FACS-sorted CD4 T cells from WT B6 mice in vitro differentiated into Th17 cells in the presence of different concentrations of SB203580 (A) or BIRB796 (B) for 72 hours. IL-17 production by Th17-polarized FACS-sorted CD4 T cells from WT B6 or MKK3 −/− MKK6 +/− mice (C) or total CD4 T cells from WT B10.BR and dn-p38-Tg (D) or MKK6 Tg (E) differentiated into Th17 cells. IL-17 levels in the supernatants were assessed by ELISA. The significance of differences observed in panels A and B was determined by linear regression analysis (A, P
    Figure Legend Snippet: p38 MAPK regulates IL-17 production by in vitro–generated Th17 cells. FACS-sorted CD4 T cells from WT B6 mice in vitro differentiated into Th17 cells in the presence of different concentrations of SB203580 (A) or BIRB796 (B) for 72 hours. IL-17 production by Th17-polarized FACS-sorted CD4 T cells from WT B6 or MKK3 −/− MKK6 +/− mice (C) or total CD4 T cells from WT B10.BR and dn-p38-Tg (D) or MKK6 Tg (E) differentiated into Th17 cells. IL-17 levels in the supernatants were assessed by ELISA. The significance of differences observed in panels A and B was determined by linear regression analysis (A, P

    Techniques Used: In Vitro, Generated, FACS, Mouse Assay, Enzyme-linked Immunosorbent Assay

    17) Product Images from "Connexin 43 Hemichannel Activity Promoted by Pro-Inflammatory Cytokines and High Glucose Alters Endothelial Cell Function"

    Article Title: Connexin 43 Hemichannel Activity Promoted by Pro-Inflammatory Cytokines and High Glucose Alters Endothelial Cell Function

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01899

    Endothelial connexin 43 (Cx43) hemichannel opening evoked by high glucose and IL-1β/TNF-α depends on Ca 2+ signaling and activation of p38 MAPK/inducible NO synthase/COX 2 -dependent pathways and EP 1 /P2 receptors: prevention by cannabinoids. (A) Averaged Etd uptake rate normalized with control conditions (5 mM glucose, dashed line) of EAhy cells treated for 72 h with 25 mM glucose and IL-1β/TNF-α alone or in combination with the following agents: 10 µM SB203580, 1 µM L-N6, 15 µM indometacin (indomet); 1 µM sc-560; 5 µM ns-398; 20 µM sc-19220, 10 µM BAPTA, 10 µM Brilliant blue G (BBG), 200 µM oxidized ATP (oATP), 10 µM MRS2179; and 10 µM A740003. * p
    Figure Legend Snippet: Endothelial connexin 43 (Cx43) hemichannel opening evoked by high glucose and IL-1β/TNF-α depends on Ca 2+ signaling and activation of p38 MAPK/inducible NO synthase/COX 2 -dependent pathways and EP 1 /P2 receptors: prevention by cannabinoids. (A) Averaged Etd uptake rate normalized with control conditions (5 mM glucose, dashed line) of EAhy cells treated for 72 h with 25 mM glucose and IL-1β/TNF-α alone or in combination with the following agents: 10 µM SB203580, 1 µM L-N6, 15 µM indometacin (indomet); 1 µM sc-560; 5 µM ns-398; 20 µM sc-19220, 10 µM BAPTA, 10 µM Brilliant blue G (BBG), 200 µM oxidized ATP (oATP), 10 µM MRS2179; and 10 µM A740003. * p

    Techniques Used: Activation Assay

    18) Product Images from "Adhesion to stromal cells mediates imatinib resistance in chronic myeloid leukemia through ERK and BMP signaling pathways"

    Article Title: Adhesion to stromal cells mediates imatinib resistance in chronic myeloid leukemia through ERK and BMP signaling pathways

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-10373-3

    ( a ) K562 cells adherent to stromal cells (MSC+) were treated with BMP inhibitor LDN193189 (LDN, 3 μM), ERK inhibitor U0126 (U01, 10 μM) and apoptosis percentage was analyzed. ( b ) Apoptosis percentage was analyzed in K562 cells overexpressing BMP2, BMP4 and treated with IM (10 µM for 48 hrs). Apoptosis percentage was analyzed in control (K) or adherent (AD-K) K562 cells treated ( c ) with NFκB inhibitor Bay 11-7082 (BAY) or P38 MAPK inhibitor SB203580 (SB) in the presence of IM for 48 hours, ( d ) CXCR4 inhibitor AMD3100 (AMD, 50 μg/ml), ( e ) BMP inhibitor LDN193189 (LDN, 3 μM), ERK inhibitor U0126 (U01, 10 μM), ROCK inhibitor Y27632 (Y27, 20 μM) and cytochalasin D (CYD, 0.3 μg/ml) along with IM (10 μM). Apoptosis percentage was determined after 48 hours of treatment. *p
    Figure Legend Snippet: ( a ) K562 cells adherent to stromal cells (MSC+) were treated with BMP inhibitor LDN193189 (LDN, 3 μM), ERK inhibitor U0126 (U01, 10 μM) and apoptosis percentage was analyzed. ( b ) Apoptosis percentage was analyzed in K562 cells overexpressing BMP2, BMP4 and treated with IM (10 µM for 48 hrs). Apoptosis percentage was analyzed in control (K) or adherent (AD-K) K562 cells treated ( c ) with NFκB inhibitor Bay 11-7082 (BAY) or P38 MAPK inhibitor SB203580 (SB) in the presence of IM for 48 hours, ( d ) CXCR4 inhibitor AMD3100 (AMD, 50 μg/ml), ( e ) BMP inhibitor LDN193189 (LDN, 3 μM), ERK inhibitor U0126 (U01, 10 μM), ROCK inhibitor Y27632 (Y27, 20 μM) and cytochalasin D (CYD, 0.3 μg/ml) along with IM (10 μM). Apoptosis percentage was determined after 48 hours of treatment. *p

    Techniques Used:

    19) Product Images from "Influence of Phosphorylation and Oligomerization on the Protective Role of the Small Heat Shock Protein 27 in Rat Adult Cardiomyocytes"

    Article Title: Influence of Phosphorylation and Oligomerization on the Protective Role of the Small Heat Shock Protein 27 in Rat Adult Cardiomyocytes

    Journal: Gene Expression

    doi:

    Western blot analysis of native polyacrylamide gel. Rat adult cardiomyocytes infected with the adenoviral construct containing the wild-type hsp27 in the absence and presence of the p38 MAP kinase inhibitor, SB203580, and the Triple G mutant of the hsp27 were either left untreated (Co) or submitted to simulated ischemia (Isc). Protein extracts from these cardiomyocytes were fractionated on a 4% native polyacrylamide gel, blotted onto nitrocellulose, and reacted with a monoclonal antibody specific to the human hsp27, as described in Materials and Methods. The position of the molecular mass markers is shown on the right side of Western blot.
    Figure Legend Snippet: Western blot analysis of native polyacrylamide gel. Rat adult cardiomyocytes infected with the adenoviral construct containing the wild-type hsp27 in the absence and presence of the p38 MAP kinase inhibitor, SB203580, and the Triple G mutant of the hsp27 were either left untreated (Co) or submitted to simulated ischemia (Isc). Protein extracts from these cardiomyocytes were fractionated on a 4% native polyacrylamide gel, blotted onto nitrocellulose, and reacted with a monoclonal antibody specific to the human hsp27, as described in Materials and Methods. The position of the molecular mass markers is shown on the right side of Western blot.

    Techniques Used: Western Blot, Infection, Construct, Mutagenesis

    Effects of p38 MAP kinase inhibition on enzyme release induced by simulated ischemia. Cardiomyocytes infected with the control (Con) and wild-type hsp27 adenoviral constructs were either left untreated or treated with the inhibitor SB203580 as described in Materials and Methods. The effect on enzyme release following simulated ischemia by the presence of the mutant Triple A was also included in this series of experiments. Data are a mean of four independent experiments. *p
    Figure Legend Snippet: Effects of p38 MAP kinase inhibition on enzyme release induced by simulated ischemia. Cardiomyocytes infected with the control (Con) and wild-type hsp27 adenoviral constructs were either left untreated or treated with the inhibitor SB203580 as described in Materials and Methods. The effect on enzyme release following simulated ischemia by the presence of the mutant Triple A was also included in this series of experiments. Data are a mean of four independent experiments. *p

    Techniques Used: Inhibition, Infection, Construct, Mutagenesis

    20) Product Images from "Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade "

    Article Title: Porins from Salmonella enterica Serovar Typhimurium Activate the Transcription Factors Activating Protein 1 and NF-?B through the Raf-1-Mitogen-Activated Protein Kinase Cascade

    Journal: Infection and Immunity

    doi: 10.1128/IAI.70.2.558-568.2002

    EMSA with AP-1 consensus oligonucleotide. U937 cells (3 × 10 6 cells/ml) were stimulated for 60 min with porins (5 μg/ml) or LPS (1 μg/ml) pretreated with inhibitors. (A) Control, control nuclear extract (5 μg); LPS, LPS-induced nuclear extract (5 μg); LPS + PD-098059, LPS-induced nuclear extract (5 μg) pretreated with PD-098059; LPS + SB202190, LPS-induced nuclear extract (5 μg) pretreated with SB203580; LPS + Forskolin, LPS-induced nuclear extract (5 μg) pretreated with forskolin; COMP., binding reactions with unlabeled excesses of AP-1 and NF-κB consensus oligonucleotides. The arrow to the left indicates control and induced complexes. (B) The same experiment using porins instead of LPS. Fold increases in AP-1 activation are shown below each lane for each blot.
    Figure Legend Snippet: EMSA with AP-1 consensus oligonucleotide. U937 cells (3 × 10 6 cells/ml) were stimulated for 60 min with porins (5 μg/ml) or LPS (1 μg/ml) pretreated with inhibitors. (A) Control, control nuclear extract (5 μg); LPS, LPS-induced nuclear extract (5 μg); LPS + PD-098059, LPS-induced nuclear extract (5 μg) pretreated with PD-098059; LPS + SB202190, LPS-induced nuclear extract (5 μg) pretreated with SB203580; LPS + Forskolin, LPS-induced nuclear extract (5 μg) pretreated with forskolin; COMP., binding reactions with unlabeled excesses of AP-1 and NF-κB consensus oligonucleotides. The arrow to the left indicates control and induced complexes. (B) The same experiment using porins instead of LPS. Fold increases in AP-1 activation are shown below each lane for each blot.

    Techniques Used: Binding Assay, Activation Assay

    21) Product Images from "A Role for p38 Mitogen-activated Protein Kinase-mediated Threonine 30-dependent Norepinephrine Transporter Regulation in Cocaine Sensitization and Conditioned Place Preference *"

    Article Title: A Role for p38 Mitogen-activated Protein Kinase-mediated Threonine 30-dependent Norepinephrine Transporter Regulation in Cocaine Sensitization and Conditioned Place Preference *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M114.612192

    In vivo p38 MAPK inhibition attenuates cocaine CPP in mice. A , cocaine CPP. Mice received 20 mg/kg intraperitoneal cocaine ( Coc ) for conditioning and were tested for CPP on day 4 following vehicle ( Veh ) or SB203580 ( SB ) injections. The CPP score is given as mean ± S.E. ( error bars ). * indicates a significant difference in cocaine CPP (one-way analysis of variance; Dunnett's test: F (4,44) = 5.31, p
    Figure Legend Snippet: In vivo p38 MAPK inhibition attenuates cocaine CPP in mice. A , cocaine CPP. Mice received 20 mg/kg intraperitoneal cocaine ( Coc ) for conditioning and were tested for CPP on day 4 following vehicle ( Veh ) or SB203580 ( SB ) injections. The CPP score is given as mean ± S.E. ( error bars ). * indicates a significant difference in cocaine CPP (one-way analysis of variance; Dunnett's test: F (4,44) = 5.31, p

    Techniques Used: In Vivo, Inhibition, Conditioned Place Preference, Mouse Assay

    Acute in vivo p38 MAPK inhibition blocks cocaine-induced NE transport and NET surface expression in the mouse PFC and NAc. Mice were injected with vehicle ( Veh ) or SB203580 ( SB ) (50 μg/kg intraperitoneally) 15 min prior to saline ( Sal ) or cocaine ( Coc ) (30 mg/kg intraperitoneally) injections. Brains were collected 1 h postinjection for PFC and NAc synaptosomal preparations. A , NE uptake. PFC and NAc synaptosomes were used for NE uptake assays as described under “Experimental Procedures.” NE uptake data derived from three separate experiments, each in triplicate, are given as mean ± S.E. ( error bars ). * indicates a significant change in NE transport (one-way analysis of variance; Dunnett's test: F (4,44) = 3.09 for PFC, p
    Figure Legend Snippet: Acute in vivo p38 MAPK inhibition blocks cocaine-induced NE transport and NET surface expression in the mouse PFC and NAc. Mice were injected with vehicle ( Veh ) or SB203580 ( SB ) (50 μg/kg intraperitoneally) 15 min prior to saline ( Sal ) or cocaine ( Coc ) (30 mg/kg intraperitoneally) injections. Brains were collected 1 h postinjection for PFC and NAc synaptosomal preparations. A , NE uptake. PFC and NAc synaptosomes were used for NE uptake assays as described under “Experimental Procedures.” NE uptake data derived from three separate experiments, each in triplicate, are given as mean ± S.E. ( error bars ). * indicates a significant change in NE transport (one-way analysis of variance; Dunnett's test: F (4,44) = 3.09 for PFC, p

    Techniques Used: In Vivo, Inhibition, Expressing, Mouse Assay, Injection, Derivative Assay

    In vivo p38 MAPK inhibition attenuates cocaine-induced locomotor sensitization, NET up-regulation, and p38 MAPK activation in mice. A , locomotor activity. Locomotor activity measured over a 1-h period on challenge day given as 10-min bin data. 50 μg/kg intraperitoneal SB203580 ( SB ) given prior to cocaine ( Coc ) challenge significantly blocked cocaine sensitization. * indicates significant changes in locomotor activity (*, p
    Figure Legend Snippet: In vivo p38 MAPK inhibition attenuates cocaine-induced locomotor sensitization, NET up-regulation, and p38 MAPK activation in mice. A , locomotor activity. Locomotor activity measured over a 1-h period on challenge day given as 10-min bin data. 50 μg/kg intraperitoneal SB203580 ( SB ) given prior to cocaine ( Coc ) challenge significantly blocked cocaine sensitization. * indicates significant changes in locomotor activity (*, p

    Techniques Used: In Vivo, Inhibition, Activation Assay, Mouse Assay, Activity Assay

    22) Product Images from "Overexpression of endophilin A1 exacerbates synaptic alterations in a mouse model of Alzheimer’s disease"

    Article Title: Overexpression of endophilin A1 exacerbates synaptic alterations in a mouse model of Alzheimer’s disease

    Journal: Nature Communications

    doi: 10.1038/s41467-018-04389-0

    Effect of EP overexpression on cerebral Aβ accumulation. ELISA for measurement of Aβ40 ( a , c ) and Aβ42 ( b , d ) in the entorhinal cortex of Tg mAPP and Tg Sh3gl2 /mAPP mice at the age of 5–5.5 months. EUK-134 (EUK, 2 mg/kg) ( c , d ) or SB203580 (SB, 0.5 mg/kg) ( c , d ) was administered to Tg Sh3gl2 /mAPP mice once a day for 3 weeks and then cortical tissues were subjected to Aβ measurement at the age of 5–5.5 months. Date are shown as mean ± s.e.m., n = 3–6 per group (one-way ANOVA in a–d ). Quantification of immunoreactive bands for Aβ ( e ), BACE1 ( g ), or IDE ( i ) in the indicated Tg mice at the age of 5–5.5 months. Quantification of immunoreactive bands for Aβ ( f ), BACE1 ( h ), or IDE ( j ) in Tg Sh3gl2 .mAPP mice treated with EUK or P38 inhibitor (SB) relative to vehicle treatment. β-Actin was used as a protein loading control. Lower panels are representative immunoblots for the indicated proteins in the indicated Tg mice. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in e–j )
    Figure Legend Snippet: Effect of EP overexpression on cerebral Aβ accumulation. ELISA for measurement of Aβ40 ( a , c ) and Aβ42 ( b , d ) in the entorhinal cortex of Tg mAPP and Tg Sh3gl2 /mAPP mice at the age of 5–5.5 months. EUK-134 (EUK, 2 mg/kg) ( c , d ) or SB203580 (SB, 0.5 mg/kg) ( c , d ) was administered to Tg Sh3gl2 /mAPP mice once a day for 3 weeks and then cortical tissues were subjected to Aβ measurement at the age of 5–5.5 months. Date are shown as mean ± s.e.m., n = 3–6 per group (one-way ANOVA in a–d ). Quantification of immunoreactive bands for Aβ ( e ), BACE1 ( g ), or IDE ( i ) in the indicated Tg mice at the age of 5–5.5 months. Quantification of immunoreactive bands for Aβ ( f ), BACE1 ( h ), or IDE ( j ) in Tg Sh3gl2 .mAPP mice treated with EUK or P38 inhibitor (SB) relative to vehicle treatment. β-Actin was used as a protein loading control. Lower panels are representative immunoblots for the indicated proteins in the indicated Tg mice. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in e–j )

    Techniques Used: Over Expression, Enzyme-linked Immunosorbent Assay, Mouse Assay, Western Blot

    Blocking EP-mediated oxidative stress and p38 activation rescued Aβ-induced synaptic vesicle recycling impairment. Fourteen-day in vitro cultured cortical neurons, either non-Tg or Tg Sh3gl2 , were treated with 50 nM Aβ for 24 h, with or without 500 nM EUK-134, 1 μM SB203580/SB, or 1 μM MitoTEMPO pretreatment for 1 h before the addition of Aβ. To visualize synaptic vesicle recycling, the cells were loaded with the fluorescent styryl dye FM1–43 before and after stimulation with 50 mM K + for the indicated time. a , f Kinetics of FM1–43 unloading of synaptic boutons during sustained stimulation with 50 mM KCl. b–e Fluorescence images before (I) and after (II, III) FM1–43 unloading with 50 mM KCl, and the representative immunofluorescence images of MAP2 (red, IV) to ensure the position of FM1–43 fluorescence (green, IV in d , e ). Tg Sh3gl2 neurons treated with 50 nM Aβ for 24 h alone ( e , h ) showed synaptic vesicle release impairment compared to the vehicle Tg Sh3gl2 treatment ( c , g ) and non-Tg neurons, whereas treatment with 50 nM Aβ ( d ) showed no difference compared to the vehicle non-Tg neurons ( b ). Pretreatment with 500 nM EUK-134 ( i ), 1 μM SB203580 ( j ), or 1 μM MitoTEMPO ( k ) rescued Aβ-induced synaptic vesicle recycling impairment in Tg Sh3gl2 neurons. Scale bar = 50 µm. Error bars represent s.e.m., n = 8 per group. * p
    Figure Legend Snippet: Blocking EP-mediated oxidative stress and p38 activation rescued Aβ-induced synaptic vesicle recycling impairment. Fourteen-day in vitro cultured cortical neurons, either non-Tg or Tg Sh3gl2 , were treated with 50 nM Aβ for 24 h, with or without 500 nM EUK-134, 1 μM SB203580/SB, or 1 μM MitoTEMPO pretreatment for 1 h before the addition of Aβ. To visualize synaptic vesicle recycling, the cells were loaded with the fluorescent styryl dye FM1–43 before and after stimulation with 50 mM K + for the indicated time. a , f Kinetics of FM1–43 unloading of synaptic boutons during sustained stimulation with 50 mM KCl. b–e Fluorescence images before (I) and after (II, III) FM1–43 unloading with 50 mM KCl, and the representative immunofluorescence images of MAP2 (red, IV) to ensure the position of FM1–43 fluorescence (green, IV in d , e ). Tg Sh3gl2 neurons treated with 50 nM Aβ for 24 h alone ( e , h ) showed synaptic vesicle release impairment compared to the vehicle Tg Sh3gl2 treatment ( c , g ) and non-Tg neurons, whereas treatment with 50 nM Aβ ( d ) showed no difference compared to the vehicle non-Tg neurons ( b ). Pretreatment with 500 nM EUK-134 ( i ), 1 μM SB203580 ( j ), or 1 μM MitoTEMPO ( k ) rescued Aβ-induced synaptic vesicle recycling impairment in Tg Sh3gl2 neurons. Scale bar = 50 µm. Error bars represent s.e.m., n = 8 per group. * p

    Techniques Used: Blocking Assay, Activation Assay, In Vitro, Cell Culture, Fluorescence, Immunofluorescence

    Inhibition of p38 MAP kinase rescues impairment on synaptic plasticity and spatial learning and memory in Tg Sh3gl2 /mAPP mice. a , b Hippocampal slices from 5-month-old to 6-month-old Tg Sh3gl2 mice were pretreated with SB203580 (SB, 1 µM) for 5 min before Aβ perfusion (100 nM for 20 min) and then hippocampal CA3-CA1 LTP was recorded ( a ). Tg Sh3gl2 /mAPP mice were intraperitoneally injected with SB203580 (0.5 mg/kg) once a day for 3 weeks and then performed LTP experiments ( b ) and Morris water maze test ( c – f ) at the age of 5–5.5 months. Upper panels of a and b show representative traces of fEPSP in the indicated slices with the indicated treatment before θ-burst stimulation (black line) after 1 h (gray line). Administration of SB203850 significantly ameliorated hippocampal LTP deficit in Tg Sh3gl2 /mAPP mice compared to the vehicle-treated group. Error bars represent s.e.m., n = 7–10 per group. * p
    Figure Legend Snippet: Inhibition of p38 MAP kinase rescues impairment on synaptic plasticity and spatial learning and memory in Tg Sh3gl2 /mAPP mice. a , b Hippocampal slices from 5-month-old to 6-month-old Tg Sh3gl2 mice were pretreated with SB203580 (SB, 1 µM) for 5 min before Aβ perfusion (100 nM for 20 min) and then hippocampal CA3-CA1 LTP was recorded ( a ). Tg Sh3gl2 /mAPP mice were intraperitoneally injected with SB203580 (0.5 mg/kg) once a day for 3 weeks and then performed LTP experiments ( b ) and Morris water maze test ( c – f ) at the age of 5–5.5 months. Upper panels of a and b show representative traces of fEPSP in the indicated slices with the indicated treatment before θ-burst stimulation (black line) after 1 h (gray line). Administration of SB203850 significantly ameliorated hippocampal LTP deficit in Tg Sh3gl2 /mAPP mice compared to the vehicle-treated group. Error bars represent s.e.m., n = 7–10 per group. * p

    Techniques Used: Inhibition, Mouse Assay, Injection

    Blocking EP-mediated oxidative stress and p38 activation rescued Aβ-induced synaptic loss. a , b Brain slices from 3-month-old non-Tg or Tg Sh3gl2 mice were perfused with Aβ (50 nM) for 2 h, and then subjected to immunoblotting analysis for synaptojanin ( a ) and synaptophysin ( b ) in the indicated groups of brain slices. β-Actin served as protein loading controls. The upper panel displays quantification of immunoreactive bands for the corresponding protein relative to β-actin. Data are expressed as fold change relative to the non-Tg vehicle control group. Data are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in a , b ). c , d The Tg Sh3gl2 brain slices from 3-month-old mice were perfused with Aβ (50 nM) for 2 h with/without pretreatment of 500 nM EUK-134 (EUK), 1 μM SB203580 (SB), or 1 µM MitoTEMPO (TEMPO) for 5 min. Immunoblotting for synaptojanin ( c ) and synaptophysin ( d ) in the indicated groups of brain slices. The upper panel displays the quantification of immunoreactive bands for the corresponding protein relative to β-actin. Data are expressed as fold change relative to Tg Sh3gl2 vehicle control group. Data are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in c , d ). Fourteen-day in vitro cultured cortical neurons, either non-Tg or Tg Sh3gl2 , were treated with 50 nM Aβ for 24 h, with or without 500 nM EUK-134, 1 μM SB203580, or 1 μM MitoTEMPO pretreatment for 1 h before the addition of Aβ. The numbers of synaptophysin-positive clusters were significantly decreased in Aβ-treated Tg Sh3gl2 neurons compared to vehicle-treated non-Tg neurons in e – h . Treatment with EUK-134, or SB203580, or MitoTEMPO, inhibited Aβ-induced synaptic loss in cultured EP overexpression neurons ( g , h ). Representative images for synaptophysin (green), MAP2 (red), and nuclei (blue) in the indicated groups of neurons are shown in e , g . Scale bars, 50 μm. Quantifications of synaptophysin-positive clusters per 10 μm of dendrites are shown in f , h . Data are shown as mean ± s.e.m., n = 12 cells for each group (one-way ANOVA in f , h )
    Figure Legend Snippet: Blocking EP-mediated oxidative stress and p38 activation rescued Aβ-induced synaptic loss. a , b Brain slices from 3-month-old non-Tg or Tg Sh3gl2 mice were perfused with Aβ (50 nM) for 2 h, and then subjected to immunoblotting analysis for synaptojanin ( a ) and synaptophysin ( b ) in the indicated groups of brain slices. β-Actin served as protein loading controls. The upper panel displays quantification of immunoreactive bands for the corresponding protein relative to β-actin. Data are expressed as fold change relative to the non-Tg vehicle control group. Data are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in a , b ). c , d The Tg Sh3gl2 brain slices from 3-month-old mice were perfused with Aβ (50 nM) for 2 h with/without pretreatment of 500 nM EUK-134 (EUK), 1 μM SB203580 (SB), or 1 µM MitoTEMPO (TEMPO) for 5 min. Immunoblotting for synaptojanin ( c ) and synaptophysin ( d ) in the indicated groups of brain slices. The upper panel displays the quantification of immunoreactive bands for the corresponding protein relative to β-actin. Data are expressed as fold change relative to Tg Sh3gl2 vehicle control group. Data are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in c , d ). Fourteen-day in vitro cultured cortical neurons, either non-Tg or Tg Sh3gl2 , were treated with 50 nM Aβ for 24 h, with or without 500 nM EUK-134, 1 μM SB203580, or 1 μM MitoTEMPO pretreatment for 1 h before the addition of Aβ. The numbers of synaptophysin-positive clusters were significantly decreased in Aβ-treated Tg Sh3gl2 neurons compared to vehicle-treated non-Tg neurons in e – h . Treatment with EUK-134, or SB203580, or MitoTEMPO, inhibited Aβ-induced synaptic loss in cultured EP overexpression neurons ( g , h ). Representative images for synaptophysin (green), MAP2 (red), and nuclei (blue) in the indicated groups of neurons are shown in e , g . Scale bars, 50 μm. Quantifications of synaptophysin-positive clusters per 10 μm of dendrites are shown in f , h . Data are shown as mean ± s.e.m., n = 12 cells for each group (one-way ANOVA in f , h )

    Techniques Used: Blocking Assay, Activation Assay, Mouse Assay, In Vitro, Cell Culture, Over Expression

    Effect of EP overexpression on p38 MAP kinase activation and mitochondrial dysfunction in Aβ-insulted brain in vivo and brain slices in vitro. a Brain slices from 3-month-old non-Tg or Tg Sh3gl2 mice were perfused with Aβ (50 nM) or vehicle for 1 h and then subjected to immunoblotting analysis for the phosphorylation of p38 MAP kinase (p-p38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Tubulin and β-actin served as a neuronal marker and protein loading controls, respectively. Data are expressed as fold change relative to the non-Tg vehicle control group. b Immunoblotting of cortical homogenates from the indicated Tg mice at 5–6 months of age for the indicated proteins. Data are expressed as fold change relative to the non-Tg mice group. c Brain slices from indicated Tg Sh3gl2 mice were treated with vehicle or Aβ (50 nM) with/without pretreatment of EUK-134 (EUK, 500 nM), SB203580 (SB, 1 µM), or mitochondrial antioxidant MitoTEMPO (TEMPO, 1 µM) for 5 min, and then subjected to immunoblotting for the phosphorylation of p38 MAP kinase (p-P38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Data are expressed as fold change relative to the Tg Sh3gl2 vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in a – c ). d Representative spectra of EPR in non-Tg and Tg Sh3gl2 brain slices with the treatment of vehicle or Aβ (50 nM) in the presence of SB203580 (1 µM). e Quantification of EPR spectra in the indicated groups of mice. f – g Mitochondrial complex IV activity ( f ) and ATP levels ( g ) in the indicated groups of brain slices treated with vehicle or Aβ in the presence/absence of SB203580. Data are expressed as fold increase relative to non-Tg vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in e – g )
    Figure Legend Snippet: Effect of EP overexpression on p38 MAP kinase activation and mitochondrial dysfunction in Aβ-insulted brain in vivo and brain slices in vitro. a Brain slices from 3-month-old non-Tg or Tg Sh3gl2 mice were perfused with Aβ (50 nM) or vehicle for 1 h and then subjected to immunoblotting analysis for the phosphorylation of p38 MAP kinase (p-p38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Tubulin and β-actin served as a neuronal marker and protein loading controls, respectively. Data are expressed as fold change relative to the non-Tg vehicle control group. b Immunoblotting of cortical homogenates from the indicated Tg mice at 5–6 months of age for the indicated proteins. Data are expressed as fold change relative to the non-Tg mice group. c Brain slices from indicated Tg Sh3gl2 mice were treated with vehicle or Aβ (50 nM) with/without pretreatment of EUK-134 (EUK, 500 nM), SB203580 (SB, 1 µM), or mitochondrial antioxidant MitoTEMPO (TEMPO, 1 µM) for 5 min, and then subjected to immunoblotting for the phosphorylation of p38 MAP kinase (p-P38), total p38 MAP kinase (t-p38), tubulin, and β-actin. Data are expressed as fold change relative to the Tg Sh3gl2 vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in a – c ). d Representative spectra of EPR in non-Tg and Tg Sh3gl2 brain slices with the treatment of vehicle or Aβ (50 nM) in the presence of SB203580 (1 µM). e Quantification of EPR spectra in the indicated groups of mice. f – g Mitochondrial complex IV activity ( f ) and ATP levels ( g ) in the indicated groups of brain slices treated with vehicle or Aβ in the presence/absence of SB203580. Data are expressed as fold increase relative to non-Tg vehicle control group. Date are shown as mean ± s.e.m., n = 3 per group (one-way ANOVA in e – g )

    Techniques Used: Over Expression, Activation Assay, In Vivo, In Vitro, Mouse Assay, Marker, Electron Paramagnetic Resonance, Activity Assay

    23) Product Images from "BCL2 inhibitor ABT-199 and JNK inhibitor SP600125 exhibit synergistic cytotoxicity against imatinib-resistant Ph+ ALL cells"

    Article Title: BCL2 inhibitor ABT-199 and JNK inhibitor SP600125 exhibit synergistic cytotoxicity against imatinib-resistant Ph+ ALL cells

    Journal: Biochemistry and Biophysics Reports

    doi: 10.1016/j.bbrep.2018.07.001

    Effects of cell signaling pathway inhibitors. (a) Western blot analysis of cellular signaling pathway proteins and their phosphorylated forms in the indicated cell lines. (b) Effects of various inhibitors on cellular signaling pathways in NphA2, NphA2/STIR, K562 and K562/DNR cells. Cells were plated at 2 × 10 5 /ml in the presence of U0126 (ERK inhibitor, 10 μM), LY294002 (AKT inhibitor, 10 μM), SB203580 (p38 inhibitor, 25 μM), or SP600125 (JNK inhibitor, 10 and 20 μM). Viable cell number was counted on day 2. Culture experiments were performed in triplicate. Data are shown as mean ± SD. (c) NphA2 and NphA2/STIR cells were cultured with or without SP600125 (20 μM) for 1 or 2 days (D1 and D2, respectively) and then examined for phosphorylated c-jun, cleaved PARP and cleaved caspase 3 expression. Because of the weaker expression of p-c-Jun of NphA2/STIR, p-c-Jun Western blotting of NphA2/STIR needed longer exposure than that of NphA2. (d) Effect of IMT on phosphorylated JNK of NphA2 cells were analyzed. Six and 24 h after IMT treatment (10 μM), cells were collected and phosphorylated JNK and total JNK expression were examined by the Western blotting. Arrowhead denotes JNK1 (lower) and JNK2 (upper), respectively.
    Figure Legend Snippet: Effects of cell signaling pathway inhibitors. (a) Western blot analysis of cellular signaling pathway proteins and their phosphorylated forms in the indicated cell lines. (b) Effects of various inhibitors on cellular signaling pathways in NphA2, NphA2/STIR, K562 and K562/DNR cells. Cells were plated at 2 × 10 5 /ml in the presence of U0126 (ERK inhibitor, 10 μM), LY294002 (AKT inhibitor, 10 μM), SB203580 (p38 inhibitor, 25 μM), or SP600125 (JNK inhibitor, 10 and 20 μM). Viable cell number was counted on day 2. Culture experiments were performed in triplicate. Data are shown as mean ± SD. (c) NphA2 and NphA2/STIR cells were cultured with or without SP600125 (20 μM) for 1 or 2 days (D1 and D2, respectively) and then examined for phosphorylated c-jun, cleaved PARP and cleaved caspase 3 expression. Because of the weaker expression of p-c-Jun of NphA2/STIR, p-c-Jun Western blotting of NphA2/STIR needed longer exposure than that of NphA2. (d) Effect of IMT on phosphorylated JNK of NphA2 cells were analyzed. Six and 24 h after IMT treatment (10 μM), cells were collected and phosphorylated JNK and total JNK expression were examined by the Western blotting. Arrowhead denotes JNK1 (lower) and JNK2 (upper), respectively.

    Techniques Used: Western Blot, Cell Culture, Expressing

    24) Product Images from "CD34− Orbital Fibroblasts From Patients With Thyroid-Associated Ophthalmopathy Modulate TNF-α Expression in CD34+ Fibroblasts and Fibrocytes"

    Article Title: CD34− Orbital Fibroblasts From Patients With Thyroid-Associated Ophthalmopathy Modulate TNF-α Expression in CD34+ Fibroblasts and Fibrocytes

    Journal: Investigative Ophthalmology & Visual Science

    doi: 10.1167/iovs.18-23951

    bTSH induced TNF-α mRNA expression is mediated through p38/MAPK. (A) Fibrocytes were pretreated with nothing or SB203580 (20 μM) for 1 hour and then treated with or without bTSH (5 mIU/mL) for 1 hour. (B) Cultures were pretreated as in (A) and then treated with or without bTSH for 45 minutes. Monolayers were solubilized and subjected to Western blot analysis for phosphorylated p38 and β-actin. Bands were quantified by densitometric analysis ([B], bottom). Data are expressed as mean ± SD of triplicates. Representative of three independent experiments using cells from a different donor.
    Figure Legend Snippet: bTSH induced TNF-α mRNA expression is mediated through p38/MAPK. (A) Fibrocytes were pretreated with nothing or SB203580 (20 μM) for 1 hour and then treated with or without bTSH (5 mIU/mL) for 1 hour. (B) Cultures were pretreated as in (A) and then treated with or without bTSH for 45 minutes. Monolayers were solubilized and subjected to Western blot analysis for phosphorylated p38 and β-actin. Bands were quantified by densitometric analysis ([B], bottom). Data are expressed as mean ± SD of triplicates. Representative of three independent experiments using cells from a different donor.

    Techniques Used: Expressing, Western Blot

    25) Product Images from "Induction of Thioredoxin Reductase 1 by Korean Red Ginseng Water Extract Regulates Cytoprotective Effects on Human Endothelial Cells"

    Article Title: Induction of Thioredoxin Reductase 1 by Korean Red Ginseng Water Extract Regulates Cytoprotective Effects on Human Endothelial Cells

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2015/972040

    Involvement of p38 and PKC- δ signaling pathway in KRG-treated TrxR1 expression in HUVECs. Cells were pretreated with 20 μ M SB203580 (p38 inhibitor) or 10 μ M Rottlerin (PKC- δ inhibitor) for 1 h, followed by incubation with 1 mg/mL KRG for 18 h. Whole cell lysates were analyzed by Western blot with antibodies against TrxR1 and GAPDH (a and c). Transient transfection of cells with 10–30 nM p38 and PKC- δ siRNA suppressed the upregulation of TrxR1 expression by KRG (b and d).
    Figure Legend Snippet: Involvement of p38 and PKC- δ signaling pathway in KRG-treated TrxR1 expression in HUVECs. Cells were pretreated with 20 μ M SB203580 (p38 inhibitor) or 10 μ M Rottlerin (PKC- δ inhibitor) for 1 h, followed by incubation with 1 mg/mL KRG for 18 h. Whole cell lysates were analyzed by Western blot with antibodies against TrxR1 and GAPDH (a and c). Transient transfection of cells with 10–30 nM p38 and PKC- δ siRNA suppressed the upregulation of TrxR1 expression by KRG (b and d).

    Techniques Used: Expressing, Incubation, Western Blot, Transfection

    26) Product Images from "Constitutive activation of MKK6 in chondrocytes of transgenic mice inhibits proliferation and delays endochondral bone formation"

    Article Title: Constitutive activation of MKK6 in chondrocytes of transgenic mice inhibits proliferation and delays endochondral bone formation

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.0507979103

    p38 MAPK increases Sox9 transcriptional activity in transient transfection and in transgenic mice. ( A ) In COS7 cells, coexpression of MKK6EE further increased the activity of the SOX9-dependent 48-bp enhancer by ≈5-fold. ( B ) Western blot analysis showed that SOX9 protein levels were not increased under these conditions. ( C ) This increase in enhancer activity was not observed in a mutant enhancer construct harboring mutations that abolish SOX9 binding. Note that in A (wild-type enhancer) and C (mutant enhancer), luciferase activity obtained in the presence of SOX9 was set to 1. The actual values of luciferase were > 100 times higher with the wild-type enhancer than with the mutant enhancer in the presence of SOX9. ( D ) In primary chondrocytes, coexpression of MKK6EE further increased the activity of the SOX9-dependent 48-bp enhancer by > 5-fold, and this increase was abolished by 10 μM of p38-specific inhibitors, SB202190, SB203580, and SB220025. In contrast, 10 μM of an inhibitor for MAPK-activated protein kinase 2 (MAPKAP2), a downstream kinase of p38, did not abolish the increased enhancer activity, suggesting that MAPKAP2 is not involved. ( E ) Northern blot analysis showed that treatment of primary chondrocytes with the p38 inhibitors decreased the expression of Col2a1 , a downstream target of Sox9, without affecting Sox9 RNA levels, further supporting the hypothesis that p38 plays a role in regulating Sox9 activity in chondrocytes. ( F ) Schematic representation of the construct used to generate transgenic mice harboring a luciferase reporter under the control of a 309-bp Col2a1 promoter and five copies of the 48-bp Col2a1 enhancer. ( G ) In mice that were homozygous for the MKK6EE transgene (MKK6EE+/+) and also heterozygous for the luciferase transgene (Luc+/-), luciferase activity in cartilage extracts showed a 2-fold increase ( P
    Figure Legend Snippet: p38 MAPK increases Sox9 transcriptional activity in transient transfection and in transgenic mice. ( A ) In COS7 cells, coexpression of MKK6EE further increased the activity of the SOX9-dependent 48-bp enhancer by ≈5-fold. ( B ) Western blot analysis showed that SOX9 protein levels were not increased under these conditions. ( C ) This increase in enhancer activity was not observed in a mutant enhancer construct harboring mutations that abolish SOX9 binding. Note that in A (wild-type enhancer) and C (mutant enhancer), luciferase activity obtained in the presence of SOX9 was set to 1. The actual values of luciferase were > 100 times higher with the wild-type enhancer than with the mutant enhancer in the presence of SOX9. ( D ) In primary chondrocytes, coexpression of MKK6EE further increased the activity of the SOX9-dependent 48-bp enhancer by > 5-fold, and this increase was abolished by 10 μM of p38-specific inhibitors, SB202190, SB203580, and SB220025. In contrast, 10 μM of an inhibitor for MAPK-activated protein kinase 2 (MAPKAP2), a downstream kinase of p38, did not abolish the increased enhancer activity, suggesting that MAPKAP2 is not involved. ( E ) Northern blot analysis showed that treatment of primary chondrocytes with the p38 inhibitors decreased the expression of Col2a1 , a downstream target of Sox9, without affecting Sox9 RNA levels, further supporting the hypothesis that p38 plays a role in regulating Sox9 activity in chondrocytes. ( F ) Schematic representation of the construct used to generate transgenic mice harboring a luciferase reporter under the control of a 309-bp Col2a1 promoter and five copies of the 48-bp Col2a1 enhancer. ( G ) In mice that were homozygous for the MKK6EE transgene (MKK6EE+/+) and also heterozygous for the luciferase transgene (Luc+/-), luciferase activity in cartilage extracts showed a 2-fold increase ( P

    Techniques Used: Activity Assay, Transfection, Transgenic Assay, Mouse Assay, Western Blot, Mutagenesis, Construct, Binding Assay, Luciferase, Northern Blot, Expressing

    27) Product Images from "Stress-Activated Protein Kinases Are Involved in Coxsackievirus B3 Viral Progeny Release"

    Article Title: Stress-Activated Protein Kinases Are Involved in Coxsackievirus B3 Viral Progeny Release

    Journal: Journal of Virology

    doi: 10.1128/JVI.79.22.13875-13881.2005

    Inhibition of p38 MAPK, but not of JNK1/2, reduces CVB3 viral progeny release. HeLa cells were preincubated with the JNK inhibitor SP600125 (A) or the p38 inhibitor SB203580 (B) for 30 min and then infected with CVB3 (MOI = 1). Twenty four hours postinfection, cell supernatants were collected, and virus titers were determined by plaque assays on HeLa cell monolayers. Values are means ± SD from three independent experiments. *, P
    Figure Legend Snippet: Inhibition of p38 MAPK, but not of JNK1/2, reduces CVB3 viral progeny release. HeLa cells were preincubated with the JNK inhibitor SP600125 (A) or the p38 inhibitor SB203580 (B) for 30 min and then infected with CVB3 (MOI = 1). Twenty four hours postinfection, cell supernatants were collected, and virus titers were determined by plaque assays on HeLa cell monolayers. Values are means ± SD from three independent experiments. *, P

    Techniques Used: Inhibition, Infection

    Inhibition of p38 MAPK does not affect CVB3 viral protein synthesis. (A) HeLa cells were preincubated with p38 inhibitor SB203580 for 30 min and then infected with CVB3 (MOI = 10) for 6 h. Cell lysates were collected and immunoblotted with antibodies against phospho-hsp27 (p-hsp27) and phospho-ATF-2. hsp27 was probed as the loading control for p-hsp27, and β-actin was probed as the loading control for p-ATF-2. Phosphorylation of hsp27 and ATF-2 was quantitated by densitometric analysis using ImageJ (National Institutes of Health, version 1.32j) and normalized to control levels (CVB3-infected cells without inhibitor) arbitrarily set to 100%. Results are means ± SD ( n = 3). *, P
    Figure Legend Snippet: Inhibition of p38 MAPK does not affect CVB3 viral protein synthesis. (A) HeLa cells were preincubated with p38 inhibitor SB203580 for 30 min and then infected with CVB3 (MOI = 10) for 6 h. Cell lysates were collected and immunoblotted with antibodies against phospho-hsp27 (p-hsp27) and phospho-ATF-2. hsp27 was probed as the loading control for p-hsp27, and β-actin was probed as the loading control for p-ATF-2. Phosphorylation of hsp27 and ATF-2 was quantitated by densitometric analysis using ImageJ (National Institutes of Health, version 1.32j) and normalized to control levels (CVB3-infected cells without inhibitor) arbitrarily set to 100%. Results are means ± SD ( n = 3). *, P

    Techniques Used: Inhibition, Infection

    28) Product Images from "Urokinase Receptor and Fibronectin Regulate the ERKMAPK to p38MAPK Activity Ratios That Determine Carcinoma Cell Proliferation or Dormancy In Vivo"

    Article Title: Urokinase Receptor and Fibronectin Regulate the ERKMAPK to p38MAPK Activity Ratios That Determine Carcinoma Cell Proliferation or Dormancy In Vivo

    Journal: Molecular Biology of the Cell

    doi:

    Effect of p38 inhibition on uPAR and α5β1 expression (A) uPAR-mRNA level. RNA extracted from D-HEp3 cells treated with DMSO (−) or 2 μM SB203580 in DMSO (+) for 5, 24, and 48 h were tested by Northern blot using 32 P-labeled uPAR-cDNA as a probe. Bottom, 36B4 mRNA used as a loading control. The graph shows the ratio of uPAR/36B4 signals, measured by laser scanning densitometry. (B) uPAR-protein level. D-HEp3 cells were incubated with 2 μM SB203580 in DMSO (+) or DMSO (−) for 5, 16, 24, or 48 h and, after lysis, the cells were processed and analyzed by Western blotting for uPAR. Total ERK served as a loading control. (C) uPAR protein levels in cells expressing a dominant negative p38. D-HEp3-neo or D-HEp3-p38DN cells were lysed and the levels of uPAR, ERK (as loading control), or p38DN were detected by Western blot. (D) Effect of Mek inhibition on SB203580-induced ERK activation and uPAR up-regulation. D-HEp3cells untreated, or treated with 2 μM SB203580 with or without 30 μM PD98059 (Mek inhibitor) for 36 h, were lysed, and the levels of uPAR protein (top) as well as active (middle) and total (bottom) ERK levels were detected by Western blot. The levels of uPAR and active ERK in T-HEp3 cells served as positive controls. (E) FACS analysis for α5β1-integrin surface expression using anti-α5β1-integrin antibodies (clone HA5) in T-HEp3, D-HEp3, or AS24 cells treated with the p38 inhibitor SB203580 (2 μM) for 48 h (SB-D-HEp3 or SB-AS24) or untreated (C-T-HEp3, C-D-HEp3, C-AS24). Isotype-matched IgG was used as control. The numbers indicate the percentages of cells positive for surface α5β1-integrin. (F) Coimmunoprecipitation of uPAR and α5β1-integrin in SB203580-treated cells. D-HEp3 cells untreated or treated with 2 μM SB203580 or 2 μM SB203580 and 30 μM PD98059 were lysed, and the cell lysates were subjected to IP with anti-α5β1-integrin antibodies and the immunoprecipitated proteins were tested by Western blotting with antibodies to β1-integrin and uPAR.
    Figure Legend Snippet: Effect of p38 inhibition on uPAR and α5β1 expression (A) uPAR-mRNA level. RNA extracted from D-HEp3 cells treated with DMSO (−) or 2 μM SB203580 in DMSO (+) for 5, 24, and 48 h were tested by Northern blot using 32 P-labeled uPAR-cDNA as a probe. Bottom, 36B4 mRNA used as a loading control. The graph shows the ratio of uPAR/36B4 signals, measured by laser scanning densitometry. (B) uPAR-protein level. D-HEp3 cells were incubated with 2 μM SB203580 in DMSO (+) or DMSO (−) for 5, 16, 24, or 48 h and, after lysis, the cells were processed and analyzed by Western blotting for uPAR. Total ERK served as a loading control. (C) uPAR protein levels in cells expressing a dominant negative p38. D-HEp3-neo or D-HEp3-p38DN cells were lysed and the levels of uPAR, ERK (as loading control), or p38DN were detected by Western blot. (D) Effect of Mek inhibition on SB203580-induced ERK activation and uPAR up-regulation. D-HEp3cells untreated, or treated with 2 μM SB203580 with or without 30 μM PD98059 (Mek inhibitor) for 36 h, were lysed, and the levels of uPAR protein (top) as well as active (middle) and total (bottom) ERK levels were detected by Western blot. The levels of uPAR and active ERK in T-HEp3 cells served as positive controls. (E) FACS analysis for α5β1-integrin surface expression using anti-α5β1-integrin antibodies (clone HA5) in T-HEp3, D-HEp3, or AS24 cells treated with the p38 inhibitor SB203580 (2 μM) for 48 h (SB-D-HEp3 or SB-AS24) or untreated (C-T-HEp3, C-D-HEp3, C-AS24). Isotype-matched IgG was used as control. The numbers indicate the percentages of cells positive for surface α5β1-integrin. (F) Coimmunoprecipitation of uPAR and α5β1-integrin in SB203580-treated cells. D-HEp3 cells untreated or treated with 2 μM SB203580 or 2 μM SB203580 and 30 μM PD98059 were lysed, and the cell lysates were subjected to IP with anti-α5β1-integrin antibodies and the immunoprecipitated proteins were tested by Western blotting with antibodies to β1-integrin and uPAR.

    Techniques Used: Inhibition, Expressing, Northern Blot, Labeling, Incubation, Lysis, Western Blot, Dominant Negative Mutation, Activation Assay, FACS, Immunoprecipitation

    Analysis of the effect of p38 inhibition on α5β1-integrin function. (A) Adhesion of DMSO- or SB203580-treated cells to FN. D-HEp3 cells were treated for 48 h with 0 or 2 μM SB203580 in DMSO, detached, and plated onto FN-coated (0.5–5 μg/ml) wells. After 10 min, the cells were fixed and stained and the attached cells were quantitated as described in MATERIALS AND METHODS. (B) a to c, confocal IF microscopy (a–c, XY sections) for FN fibrils in D-HEp3 cells treated with DMSO alone (a), 2 μM SB203580 (b), or SB203580 with 5 μg/ml human FN (c). Bar, 40 μm. d to f, effect on FN fibril formation of transient transfection of uPAR in D-HEp3. Cells transfected with empty vector (d) or with the vector encoding uPAR (e and f) and grown in serum-containing medium were fixed and stained for FN. Note that only cells transfected with uPAR are able to organize FN into thick bundles and fibrils. A larger magnification of the fibrils is shown in f. Bar, 20 μm. The arrows indicate fibrils. (C) Quantitation of the effect of p38 inhibition on FN fibrillogenesis. Cells (D-HEp3 or AS24) untreated (control) or treated with SB203580 (SB 2 μM) were incubated for 48 h with or without 5 μg/ml FN (FN and SB 2 μM + FN). Cells positive for FN fibrils are shown as percentages of cells with DAPI-positive nuclei.
    Figure Legend Snippet: Analysis of the effect of p38 inhibition on α5β1-integrin function. (A) Adhesion of DMSO- or SB203580-treated cells to FN. D-HEp3 cells were treated for 48 h with 0 or 2 μM SB203580 in DMSO, detached, and plated onto FN-coated (0.5–5 μg/ml) wells. After 10 min, the cells were fixed and stained and the attached cells were quantitated as described in MATERIALS AND METHODS. (B) a to c, confocal IF microscopy (a–c, XY sections) for FN fibrils in D-HEp3 cells treated with DMSO alone (a), 2 μM SB203580 (b), or SB203580 with 5 μg/ml human FN (c). Bar, 40 μm. d to f, effect on FN fibril formation of transient transfection of uPAR in D-HEp3. Cells transfected with empty vector (d) or with the vector encoding uPAR (e and f) and grown in serum-containing medium were fixed and stained for FN. Note that only cells transfected with uPAR are able to organize FN into thick bundles and fibrils. A larger magnification of the fibrils is shown in f. Bar, 20 μm. The arrows indicate fibrils. (C) Quantitation of the effect of p38 inhibition on FN fibrillogenesis. Cells (D-HEp3 or AS24) untreated (control) or treated with SB203580 (SB 2 μM) were incubated for 48 h with or without 5 μg/ml FN (FN and SB 2 μM + FN). Cells positive for FN fibrils are shown as percentages of cells with DAPI-positive nuclei.

    Techniques Used: Inhibition, Staining, Microscopy, Transfection, Plasmid Preparation, Quantitation Assay, Incubation

    Analysis of cross-talk between ERK and p38 pathways. (A) Effect of p38 inhibition on ERK activation (short-term treatment). Serum-starved T-HEp3 (left) or D-HEp3 (right) cells were treated for 5 or 20 min with 0, 2, and 5 μM SB203580 in DMSO, then lysed, and assayed by Western blotting using anti-phospho-ERK for active ERK or anti-ERK antibodies for total ERK. (B) Long-term treatment. D-HEp3 cells were treated in the absence of serum for 5, 24, and 48 h with 2 μM SB203580 (+) or DMSO (−) and assayed after lysis for ERK activation as indicated in A. After 48 h of treatment some cultures were washed free of the inhibitor and incubated for an additional 24 and 72 h in serum-free medium (lanes indicated as wash-out) before lysing and testing for active ERK. (C) AS24 cells were treated as in B: DMSO treated (−) for 5 or 24 h or SB203580 treated (+) for 5, 24, and 48 h; some plates were treated with the inhibitor for 48 h, washed, and incubated in serum-free medium for an additional 24 h (wash out). (D) Effect of the stable expression of a dominant negative p38 on ERK activation. Pools of D-HEp3-neo or D-HEp3-p38DN cells were lysed and the levels of active and total ERK, as well as the levels of FLAG protein, were detected by Western blotting as in A and compared with the level of active ERK in parental, tumorigenic T-HEp3 cells.
    Figure Legend Snippet: Analysis of cross-talk between ERK and p38 pathways. (A) Effect of p38 inhibition on ERK activation (short-term treatment). Serum-starved T-HEp3 (left) or D-HEp3 (right) cells were treated for 5 or 20 min with 0, 2, and 5 μM SB203580 in DMSO, then lysed, and assayed by Western blotting using anti-phospho-ERK for active ERK or anti-ERK antibodies for total ERK. (B) Long-term treatment. D-HEp3 cells were treated in the absence of serum for 5, 24, and 48 h with 2 μM SB203580 (+) or DMSO (−) and assayed after lysis for ERK activation as indicated in A. After 48 h of treatment some cultures were washed free of the inhibitor and incubated for an additional 24 and 72 h in serum-free medium (lanes indicated as wash-out) before lysing and testing for active ERK. (C) AS24 cells were treated as in B: DMSO treated (−) for 5 or 24 h or SB203580 treated (+) for 5, 24, and 48 h; some plates were treated with the inhibitor for 48 h, washed, and incubated in serum-free medium for an additional 24 h (wash out). (D) Effect of the stable expression of a dominant negative p38 on ERK activation. Pools of D-HEp3-neo or D-HEp3-p38DN cells were lysed and the levels of active and total ERK, as well as the levels of FLAG protein, were detected by Western blotting as in A and compared with the level of active ERK in parental, tumorigenic T-HEp3 cells.

    Techniques Used: Inhibition, Activation Assay, Western Blot, Lysis, Incubation, Expressing, Dominant Negative Mutation

    Effect of p38 inhibition on uPAR and β1-integrin surface expression. Untreated T-HEp3 cells (A–C) or D-HEp3 cells treated for 48 h with 2 μM SB203580 (G–I) or DMSO alone (D–F) were fixed and stained, without permeabilization, for β1-integrin (A, D, and G, AIIB2 antibody, green) or uPAR (B, E, H, R2 antibody, red), as described in MATERIALS AND METHODS, and detected by confocal laser scanning IF microscopy. C, F, and I, show the merged image of the uPAR and β1-integrin signals; yellow indicates colocalization of uPAR and β1-integrin signals. A to I are XY sections; insets in C, F, and I are XZ sections of the same cells showing the overlay. Note the high surface (apical and basolateral) expression of uPAR in D-HEp3 cells treated with the p38 inhibitor (compare E with H) that colocalizes with β1-integrin signal (compare F with I and the corresponding insets), which remains unchanged in SB203580-treated cells (compare D and G). Bars, 40 μm.
    Figure Legend Snippet: Effect of p38 inhibition on uPAR and β1-integrin surface expression. Untreated T-HEp3 cells (A–C) or D-HEp3 cells treated for 48 h with 2 μM SB203580 (G–I) or DMSO alone (D–F) were fixed and stained, without permeabilization, for β1-integrin (A, D, and G, AIIB2 antibody, green) or uPAR (B, E, H, R2 antibody, red), as described in MATERIALS AND METHODS, and detected by confocal laser scanning IF microscopy. C, F, and I, show the merged image of the uPAR and β1-integrin signals; yellow indicates colocalization of uPAR and β1-integrin signals. A to I are XY sections; insets in C, F, and I are XZ sections of the same cells showing the overlay. Note the high surface (apical and basolateral) expression of uPAR in D-HEp3 cells treated with the p38 inhibitor (compare E with H) that colocalizes with β1-integrin signal (compare F with I and the corresponding insets), which remains unchanged in SB203580-treated cells (compare D and G). Bars, 40 μm.

    Techniques Used: Inhibition, Expressing, Staining, Microscopy

    29) Product Images from "Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury"

    Article Title: Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury

    Journal: Experimental neurology

    doi: 10.1016/j.expneurol.2008.05.025

    Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p
    Figure Legend Snippet: Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p

    Techniques Used: Inhibition, Activation Assay, Activity Assay, Produced

    Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p
    Figure Legend Snippet: Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p

    Techniques Used: Inhibition, Activation Assay, Injection

    30) Product Images from "Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury"

    Article Title: Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury

    Journal: Experimental neurology

    doi: 10.1016/j.expneurol.2008.05.025

    Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p
    Figure Legend Snippet: Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p

    Techniques Used: Inhibition, Activation Assay, Activity Assay, Produced

    Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p
    Figure Legend Snippet: Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p

    Techniques Used: Inhibition, Activation Assay, Injection

    31) Product Images from "Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury"

    Article Title: Activation of p38 MAP Kinase is Involved in Central Neuropathic Pain Following Spinal Cord Injury

    Journal: Experimental neurology

    doi: 10.1016/j.expneurol.2008.05.025

    Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p
    Figure Legend Snippet: Inhibition of p38 MAPK activation significantly decreases background activity and evoked neuronal responses to brush, press, pinch, and graded von Frey stimulation at the level of SCI. A-D Spike frequency histograms of neuronal activity to brush, press, pinch and von Frey stimuli prior to SB203580 administration, and at 15, 30, 60, and 120 minutes after vehicle (A), 0.1 uM (B), 1 uM (C), or 10 uM (D) SB203580 administration. Mean activity (± SEM) for background and responses to brush (E), press (F), and pinch (G) for sham (open symbols) and SCI (filled symbols) rats given vehicle or SB203580 over the 120 minute recording period. SB203580 not only significantly depressed background activity but also dose dependently produced significant decreases in neuronal responding to brush, press, and pinch (*p

    Techniques Used: Inhibition, Activation Assay, Activity Assay, Produced

    Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p
    Figure Legend Snippet: Inhibition of p38 MAPK activation reverses at-level mechanical allodynia after SCI. The inhibitor of p38 MAPK activation, SB203580, dose dependently reversed at-level mechanical allodynia when subjects are tested at 35 days post injury. The percentage of supraspinal nociceptive responses (y axis) made by sham (Sham), and spinal injured rats (SCI) tested is graphed. Prior to SB203580 injection or vehicle, SCI rats displayed at-level mechanical allodynia (as evidenced by an increased percentage of supraspinal nociceptive responses). SB203580 dose dependently reversed this at-level mechanical allodynia to both von Frey (panel A, * p

    Techniques Used: Inhibition, Activation Assay, Injection

    32) Product Images from "β, β-Dimethylacrylshikonin induces mitochondria-dependent apoptosis of human lung adenocarcinoma cells in vitro via p38 pathway activation"

    Article Title: β, β-Dimethylacrylshikonin induces mitochondria-dependent apoptosis of human lung adenocarcinoma cells in vitro via p38 pathway activation

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2014.108

    DMAS-induced A549 cell apoptosis is mediated through p38 activation. (A) A549 cells were pretreated with or without SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measured as controls. (B) A549 cells were pretreated or not with SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (C) A549 cells were pretreated or not with SP600125 (25 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (D) A549 cells were treated with DMAS in the presence or absence of SB203580 (10 μmol/L) for 24 h. The apoptotic status was determined using an Annexin V-FITC binding assay. (E) A549 cells were pretreated with or without Z-VAD-FMK (10 μmol/L), followed by treatment with DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measure as controls.
    Figure Legend Snippet: DMAS-induced A549 cell apoptosis is mediated through p38 activation. (A) A549 cells were pretreated with or without SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measured as controls. (B) A549 cells were pretreated or not with SB203580 (10 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (C) A549 cells were pretreated or not with SP600125 (25 μmol/L), followed by treatment with DMSO (vehicle) or DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis using antibodies against cleaved caspase-3 and cleaved PARP. Actin protein levels were also measured as controls. (D) A549 cells were treated with DMAS in the presence or absence of SB203580 (10 μmol/L) for 24 h. The apoptotic status was determined using an Annexin V-FITC binding assay. (E) A549 cells were pretreated with or without Z-VAD-FMK (10 μmol/L), followed by treatment with DMAS (15 μmol/L) for 24 h. Protein extracts were prepared and subjected to Western blot analysis to measure the levels of phosphorylated p38. Total p38 protein levels were also measure as controls.

    Techniques Used: Activation Assay, Western Blot, Binding Assay

    33) Product Images from "The Frizzled-ligand Norrin acts as a tumour suppressor linking oncogenic RAS signalling to p53"

    Article Title: The Frizzled-ligand Norrin acts as a tumour suppressor linking oncogenic RAS signalling to p53

    Journal: bioRxiv

    doi: 10.1101/666925

    Norrin expression is induced by the RAS-RAF pathway. a. Norrin mRNA is expressed at high levels in transformation prone TIG3-T cells (pRS RB, Small T antigene (ST) and H-RASV12) as compared to TIG3-T control cells. Norrin mRNA levels were measured by RT-QPCR. b. Oncogenic H-RASV12 induces Norrin expression. TIG3-T cells were infected either with LMP pRB, pBabeST, shRB or pBabe H-RASV12. Norrin mRNA levels were determined by RT-QPCR. c. Norrin mRNA is induced by BRAF. mRNA levels of Norrin mRNA determined by RT-QPCR of mRNA harvested from TIG3-T cells expressing BRAF-ER. BRAF activity was induced by OHT treatment for the indicated times (h= hours). d. The MEK/ERK pathway activity is required for H-RASV12 induced expression of Norrin. RT-QPCR analysis of Norrin mRNA levels from TIG3-T cells infected with H-RASV12 expressing retrovirus and treated with the indicated inhibitors for 24 hours. U0126: MEK inhibitor; PD98059: ERK inhibitor; Wortmannin: PI3K inhibitor; SB203580: p38/MAPK inhibitor. e. The MEK-ERK pathway activity is required for the induction of Norrin expression by BRAF. RT-QPCR analysis of Norrin mRNA from TIG3-T BRAF-ER cells treated for 24h with OHT plus the indicated inhibitor. f. Illustration of the Norrin-p53 pathway with the inhibitors used in panels d,e indicated. Norrin provides a novel molecular link between RAS-RAF-MEK-ERK oncogenic signalling and p53, through activation of the WNT/β-catenin pathway.
    Figure Legend Snippet: Norrin expression is induced by the RAS-RAF pathway. a. Norrin mRNA is expressed at high levels in transformation prone TIG3-T cells (pRS RB, Small T antigene (ST) and H-RASV12) as compared to TIG3-T control cells. Norrin mRNA levels were measured by RT-QPCR. b. Oncogenic H-RASV12 induces Norrin expression. TIG3-T cells were infected either with LMP pRB, pBabeST, shRB or pBabe H-RASV12. Norrin mRNA levels were determined by RT-QPCR. c. Norrin mRNA is induced by BRAF. mRNA levels of Norrin mRNA determined by RT-QPCR of mRNA harvested from TIG3-T cells expressing BRAF-ER. BRAF activity was induced by OHT treatment for the indicated times (h= hours). d. The MEK/ERK pathway activity is required for H-RASV12 induced expression of Norrin. RT-QPCR analysis of Norrin mRNA levels from TIG3-T cells infected with H-RASV12 expressing retrovirus and treated with the indicated inhibitors for 24 hours. U0126: MEK inhibitor; PD98059: ERK inhibitor; Wortmannin: PI3K inhibitor; SB203580: p38/MAPK inhibitor. e. The MEK-ERK pathway activity is required for the induction of Norrin expression by BRAF. RT-QPCR analysis of Norrin mRNA from TIG3-T BRAF-ER cells treated for 24h with OHT plus the indicated inhibitor. f. Illustration of the Norrin-p53 pathway with the inhibitors used in panels d,e indicated. Norrin provides a novel molecular link between RAS-RAF-MEK-ERK oncogenic signalling and p53, through activation of the WNT/β-catenin pathway.

    Techniques Used: Expressing, Transformation Assay, Quantitative RT-PCR, Infection, Activity Assay, Activation Assay

    34) Product Images from "Oxidative Stress-Induced Intestinal Epithelial Cell Apoptosis is Mediated By p38 Mapk"

    Article Title: Oxidative Stress-Induced Intestinal Epithelial Cell Apoptosis is Mediated By p38 Mapk

    Journal: Biochemical and biophysical research communications

    doi: 10.1016/j.bbrc.2006.09.103

    Effect of SB203580, a specific p38 inhibitor, on H 2 O 2 -induced RIE-1 cell death RIE-1 cells were pretreated with the p38 inhibitor, SB203580 (10 μM), for 30 min prior to H 2 O 2 treatment (500 μM) for 3 h. (A) Apoptosis was estimated by an ELISA assay (Data represent triplicate determinations; mean ± SEM; * = p
    Figure Legend Snippet: Effect of SB203580, a specific p38 inhibitor, on H 2 O 2 -induced RIE-1 cell death RIE-1 cells were pretreated with the p38 inhibitor, SB203580 (10 μM), for 30 min prior to H 2 O 2 treatment (500 μM) for 3 h. (A) Apoptosis was estimated by an ELISA assay (Data represent triplicate determinations; mean ± SEM; * = p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    p38 MAPK activation is upstream of PKC activation in H 2 O 2 -induced RIE-1 cell death (A) RIE-1 cells were treated with H 2 O 2 to determine expression of phospho-PKC (pan) and β-actin by immunoblotting. (B) RIE-1 cells were pretreated with the p38 inhibitor SB203580 (10 μM) for 30 min prior to H 2 O 2 (500 μM) and phospho-PKC (pan) and β-actin were determined as above. (C) RIE-1 cells were pretreated with the PKC inhibitor GF109203x (5 μM) for 30 min prior to treatment with H 2 O 2 (500 μM) for 3 h. Western blotting performed using anti-phospho-p38 and anti-β-actin. Representative blots of at least three repeats at all data points are shown.
    Figure Legend Snippet: p38 MAPK activation is upstream of PKC activation in H 2 O 2 -induced RIE-1 cell death (A) RIE-1 cells were treated with H 2 O 2 to determine expression of phospho-PKC (pan) and β-actin by immunoblotting. (B) RIE-1 cells were pretreated with the p38 inhibitor SB203580 (10 μM) for 30 min prior to H 2 O 2 (500 μM) and phospho-PKC (pan) and β-actin were determined as above. (C) RIE-1 cells were pretreated with the PKC inhibitor GF109203x (5 μM) for 30 min prior to treatment with H 2 O 2 (500 μM) for 3 h. Western blotting performed using anti-phospho-p38 and anti-β-actin. Representative blots of at least three repeats at all data points are shown.

    Techniques Used: Activation Assay, Expressing, Western Blot

    35) Product Images from "Oxidative Stress-Induced Intestinal Epithelial Cell Apoptosis is Mediated By p38 Mapk"

    Article Title: Oxidative Stress-Induced Intestinal Epithelial Cell Apoptosis is Mediated By p38 Mapk

    Journal: Biochemical and biophysical research communications

    doi: 10.1016/j.bbrc.2006.09.103

    Effect of SB203580, a specific p38 inhibitor, on H 2 O 2 -induced RIE-1 cell death RIE-1 cells were pretreated with the p38 inhibitor, SB203580 (10 μM), for 30 min prior to H 2 O 2 treatment (500 μM) for 3 h. (A) Apoptosis was estimated by an ELISA assay (Data represent triplicate determinations; mean ± SEM; * = p
    Figure Legend Snippet: Effect of SB203580, a specific p38 inhibitor, on H 2 O 2 -induced RIE-1 cell death RIE-1 cells were pretreated with the p38 inhibitor, SB203580 (10 μM), for 30 min prior to H 2 O 2 treatment (500 μM) for 3 h. (A) Apoptosis was estimated by an ELISA assay (Data represent triplicate determinations; mean ± SEM; * = p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    p38 MAPK activation is upstream of PKC activation in H 2 O 2 -induced RIE-1 cell death (A) RIE-1 cells were treated with H 2 O 2 to determine expression of phospho-PKC (pan) and β-actin by immunoblotting. (B) RIE-1 cells were pretreated with the p38 inhibitor SB203580 (10 μM) for 30 min prior to H 2 O 2 (500 μM) and phospho-PKC (pan) and β-actin were determined as above. (C) RIE-1 cells were pretreated with the PKC inhibitor GF109203x (5 μM) for 30 min prior to treatment with H 2 O 2 (500 μM) for 3 h. Western blotting performed using anti-phospho-p38 and anti-β-actin. Representative blots of at least three repeats at all data points are shown.
    Figure Legend Snippet: p38 MAPK activation is upstream of PKC activation in H 2 O 2 -induced RIE-1 cell death (A) RIE-1 cells were treated with H 2 O 2 to determine expression of phospho-PKC (pan) and β-actin by immunoblotting. (B) RIE-1 cells were pretreated with the p38 inhibitor SB203580 (10 μM) for 30 min prior to H 2 O 2 (500 μM) and phospho-PKC (pan) and β-actin were determined as above. (C) RIE-1 cells were pretreated with the PKC inhibitor GF109203x (5 μM) for 30 min prior to treatment with H 2 O 2 (500 μM) for 3 h. Western blotting performed using anti-phospho-p38 and anti-β-actin. Representative blots of at least three repeats at all data points are shown.

    Techniques Used: Activation Assay, Expressing, Western Blot

    36) Product Images from "Inhibition of NF-?B-dependent Transcription by MKP-1"

    Article Title: Inhibition of NF-?B-dependent Transcription by MKP-1

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M109.028381

    Effect of the p38 MAPK inhibitors, SB203580 and SB239063, on NF-κB-dependent transcription and IL-8 expression. A , BEAS-2B and A549 cells were incubated with various concentrations of SB203580, SB239063, or SB202474 as indicated for 30 min prior
    Figure Legend Snippet: Effect of the p38 MAPK inhibitors, SB203580 and SB239063, on NF-κB-dependent transcription and IL-8 expression. A , BEAS-2B and A549 cells were incubated with various concentrations of SB203580, SB239063, or SB202474 as indicated for 30 min prior

    Techniques Used: Expressing, Incubation

    37) Product Images from "DUSP1 Maintains IRF1 and Leads to Increased Expression of IRF1-dependent Genes"

    Article Title: DUSP1 Maintains IRF1 and Leads to Increased Expression of IRF1-dependent Genes

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.728964

    Effect of MAPK inhibitors on IL1B-induced CXCL10 expression. A , A549 cells were either not stimulated, treated with IL1B (1 ng/ml), or pretreated with a combination of UO126 and SB203580 plus JNK inhibitor 8 each at 10 μ m ( SB + UO + J8 ) for 30 min
    Figure Legend Snippet: Effect of MAPK inhibitors on IL1B-induced CXCL10 expression. A , A549 cells were either not stimulated, treated with IL1B (1 ng/ml), or pretreated with a combination of UO126 and SB203580 plus JNK inhibitor 8 each at 10 μ m ( SB + UO + J8 ) for 30 min

    Techniques Used: Expressing

    Effect of MAPK inhibitors on IRF1 expression. A , A549 cells were either not treated ( NT ), treated with IL1B (1 ng/ml), or pretreated with either UO126, SB203580, JNK inhibitor 8 ( JNK-IN-8 ), or a combination of UO126 and SB203580 plus JNK-IN-8 each at
    Figure Legend Snippet: Effect of MAPK inhibitors on IRF1 expression. A , A549 cells were either not treated ( NT ), treated with IL1B (1 ng/ml), or pretreated with either UO126, SB203580, JNK inhibitor 8 ( JNK-IN-8 ), or a combination of UO126 and SB203580 plus JNK-IN-8 each at

    Techniques Used: Expressing

    38) Product Images from "Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication"

    Article Title: Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication

    Journal: Journal of Virology

    doi: 10.1128/JVI.75.6.2710-2728.2001

    Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.
    Figure Legend Snippet: Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.

    Techniques Used: Inhibition, Activity Assay, Incubation, Infection, Transfection

    Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.
    Figure Legend Snippet: Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.

    Techniques Used: Infection, Activity Assay, Pulse Chase, Labeling, Cell Culture, Immunoprecipitation, Autoradiography, Kinase Assay

    39) Product Images from "Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication"

    Article Title: Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication

    Journal: Journal of Virology

    doi: 10.1128/JVI.75.6.2710-2728.2001

    Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.
    Figure Legend Snippet: Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.

    Techniques Used: Inhibition, Activity Assay, Incubation, Infection, Transfection

    Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.
    Figure Legend Snippet: Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.

    Techniques Used: Infection, Activity Assay, Pulse Chase, Labeling, Cell Culture, Immunoprecipitation, Autoradiography, Kinase Assay

    40) Product Images from "Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication"

    Article Title: Herpes Simplex Virus Type 1 Blocks the Apoptotic Host Cell Defense Mechanisms That Target Bcl-2 and Manipulates Activation of p38 Mitogen-Activated Protein Kinase To Improve Viral Replication

    Journal: Journal of Virology

    doi: 10.1128/JVI.75.6.2710-2728.2001

    Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.
    Figure Legend Snippet: Inhibition of p38MAPK decreases levels of viral proteins and transcriptional activity of specific promoters. (A) Cells (10 6 ) were incubated with 30 μM SB203580 or DMSO for 1 h and infected with wt HSV-1 for 6 to 12 h in the continuous presence of SB203580 or DMSO. Immunoblotting of total cell extracts was performed for viral proteins ICP0 (upper panel) and ICP4 (lower panel) at the time points indicated. (B) Cells (10 6 ) were treated with 30 μM SB203580 or DMSO as described for panel A and infected with wt HSV-1. At 9 h postinfection, total cell extracts were analyzed for viral proteins ICP27 (upper panel) and UL42 (lower panel) by immunoblotting. (C) Cells (5 × 10 6 ) were transfected with 3 μg of pIE1CAT or pIE3CAT. At 30 h posttransfection, cells were pretreated for 1 h with DMSO or SB203580 and infected with wt HSV-1 in the continuous presence of DMSO or SB203580. CAT activity was measured at 16 h postinfection.

    Techniques Used: Inhibition, Activity Assay, Incubation, Infection, Transfection

    Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.
    Figure Legend Snippet: Infection of cells with 27lacZ but not with the wt virus induces a p38MAPK-dependent decrease in Bcl-2 half-life, and p38MAPK kinase activity coimmunoprecipitates with Bcl-2 in cells infected with ts k and 27lacZ. (A) Left panel [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with wt HSV-1. Cells (5 × 10 6 ) were infected with wt HSV-1 and at 7 h post-infection cells were labeled with [ 35 S]methionine and either lysed (0 h) or cultured in medium containing unlabeled methionine prior to immunoprecipitation for Bcl-2. The gel was analyzed by autoradiography. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ. Cells (5 × 10 6 ) were infected with 27lacZ, and at 7 h postinfection cells were analyzed as described for panel A. (B) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with wt HSV-1 and 27lacZ as described for panel A. (C) Left panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of DMSO. Cells (5 × 10 6 ) were treated with 30 μM DMSO for 1 h and infected with 27lacZ in the continuous presence of DMSO. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analysed as described for panel A. Right panel, [ 35 S]methionine pulse-chase analysis of Bcl-2 degradation rates during infection with 27lacZ in the presence of SB203580. Cells (5 × 10 6 ) were treated with 30 μM SB203580 for 1 h and infected with 27lacZ in the continuous presence of SB203580. At 7 h postinfection cells were labeled with [ 35 S]methionine, and Bcl-2 levels were analyzed as described for panel A. Preimmune serum (pre-imm.) was used at 0 h postlabeling. (D) Quantification of Bcl-2 levels from [ 35 S]methionine pulse-chase analysis during infection with 27lacZ in the presence of DMSO or SB203580, as described for panel C. (E) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h and mock infected or infected with ts k in the continuous presence of SB203580 or DMSO. In the upper panel, Bcl-2 was immunoprecipitated from total cell extracts and associated p38MAPK kinase activity was detected in immunocomplex kinase assays, using 1 μg of ATF-2 protein as the substrate. In the lower panel, total cell extracts from duplicate preparations similar to the ones analyzed in panel E, upper panel, were immunoblotted for Bcl-2. (F) Cells (10 6 ) were pretreated with 30 μM SB203580 or DMSO for 1 h, and mock infected or infected with 27lacZ, and analyzed as described for panel E. Upper panel, immunocomplex kinase assay for p38MAPK activity. Lower panel, immunoblotting of duplicate preparations for Bcl-2.

    Techniques Used: Infection, Activity Assay, Pulse Chase, Labeling, Cell Culture, Immunoprecipitation, Autoradiography, Kinase Assay

    Related Articles

    MTT Assay:

    Article Title: Inhibition of Cell Survival by Curcumin Is Associated with Downregulation of Cell Division Cycle 20 (Cdc20) in Pancreatic Cancer Cells
    Article Snippet: .. The anti-β-actin, anti-Tubulin, and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). .. Inhibitor of APC/C (Apcin), a Cdc20 inhibitor, was purchased from BostonBiochem (Boston, MA, USA).

    Injection:

    Article Title: An improved model for the binding of lidocaine and structurally related local anaesthetics to fast-inactivated voltage-operated sodium channels, showing evidence of cooperativity
    Article Snippet: .. Lidocaine and 4-chlorophenol (Sigma Chemicals, Deisenhofen, Germany) were injected directly into the bath solution before the experiments; 2,6-dimethylphenol (FLUKA, Deisenhofen, Germany) and 3,5-dimethyl-4-chlorophenol (Sigma Chemicals, Deisenhofen, Germany) were prepared as 1 M stock solutions in ethanol, light protected, and stored in glass vessels at −20°C. ..

    other:

    Article Title: Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics
    Article Snippet: Hydroxynonenal-dimethyl acetate, sodium azide, dibromobimane (DBB), dimethylsulfoxide (DMSO) and diamide were from Sigma.

    Concentration Assay:

    Article Title: Reduction of post injury neointima formation due to 17?-estradiol and phytoestrogen treatment is not influenced by the pure synthetic estrogen receptor antagonist ICI 182,780 in vitro
    Article Snippet: .. Control groups were held in medium containing 1 % isopropanol (Roth, Karlsruhe, Germany) and 1% dimethyl sulphoxide (DMSO) (Sigma, Deisenhofen, Germany) because 17β-estradiol, Genistein and Daidzein were dissolved in isopropanol and DMSO of the same concentration. .. All aortic rings were held separately in six-well plates for 21 days at 37°C with phenol red free Dulbecco's modified Eagle medium (DMEM) with Ham's F12 (mixed 1 plus 4; Gibco, Eggenstein, Germany), containing D-glucose (4.5 g/l), 15 % fetal calf serum (fcs) (Bio Whittacker, Heidelberg, Germany) and 2,5 ml/l Penicillin-Streptomycin (Gibco, Eggenstein, Germany).

    Article Title: The type III intermediate filament vimentin regulates organelle distribution and modulates autophagy
    Article Snippet: .. Chemicals Withaferin-A (WFA) (Tocris, Abingdon, UK) was diluted in dimethyl sulphoxide (DMSO) (Sigma-Aldrich, Irvine, UK) to a stock concentration of 1mg/ml. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore p38 inhibitor sb203580
    Role of <t>p38</t> in focal adhesion disassembly and cytoskeletal arrangement regulation JC cells were treated with 15d-PGJ 2 (0.3 μM, 15 and 30 min) or ethanol (EtOH) as a vehicle control and phosphorylated p38 (p-p38) was determined by Western blot analysis and quantified. A representative Western blot image is shown. Values represent the ratio of p-p38/total p38 normalized to time-matched vehicle control (A). JC cells were pretreated with the p38 inhibitor <t>SB203580</t> (“SB”; 10 μM, 30 min), and then 15d-PGJ 2 (0.3 μM) was added for an additional 4 h. Cells were fixed in 3% glutaraldehyde and focal adhesions quantified using interference reflection microscopy. Values represent the mean percentage of cells scored positive for focal adhesions (B). JC cells were also pretreated with SB (10 μM, 30 min), and then BD-15d-PGJ 2 (0.24 μM) was added for an additional 30 min. Cells were then fixed, permeabilized and stained with 2 units of Alexa Fluor® 633 Phalloidin to visualize F-actin (red channel). Nuclei were visualized with DAPI (blue channel). Representative images of red and blue channel merged images are shown from samples prepared in triplicate. EtOH and DMSO were used as vehicle controls. (C). Values shown represent means ± SEM, n = at least 3-6. ** p
    P38 Inhibitor Sb203580, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1063 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p38 inhibitor sb203580/product/Millipore
    Average 99 stars, based on 1063 article reviews
    Price from $9.99 to $1999.99
    p38 inhibitor sb203580 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    88
    Millipore sb203508
    Involvement of p38 MAPK in ethanol induction of ISRE transcription and Stat1 serine phosphorylation. Panel A, Huh7 cells were transfected with 0.7 μg pISRE-luc, and 22 hours later, cells were incubated for 2 hours at the indicated μM concentrations of <t>SB203508</t> (a p38 inhibitor), followed by 100 mM ethanol. Cell lysates were harvested 6 hours later and luciferase results were normalized to amounts of total cellular protein. Error bars represent standard deviations. The experiment was repeated 4 times with identical results. B, Huh7 cells were treated for 2 hours in the presence of 50 μM of various MAPK inhibitors, and stimulated with 100 mM alcohol for 20 minutes. Whole cell protein extracts were blotted for the serine phosphorylated form (S727) or total form of Stat1. The experiment was repeated twice, yielding similar results. C, Huh7 cells were transfected with control vector plasmid (Vec) or a plasmid expressing a dominant negative mutant p38 protein (p38 AGF). Twenty-four hours later, cells were not treated or treated for 20 minutes with 100 mM ethanol. Levels of S727 and total Stat1 and transfected p38 proteins were determined by western blot. The figure is representative of 2 independent experiments that produced similar results.
    Sb203508, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sb203508/product/Millipore
    Average 88 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    sb203508 - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

    Image Search Results


    Role of p38 in focal adhesion disassembly and cytoskeletal arrangement regulation JC cells were treated with 15d-PGJ 2 (0.3 μM, 15 and 30 min) or ethanol (EtOH) as a vehicle control and phosphorylated p38 (p-p38) was determined by Western blot analysis and quantified. A representative Western blot image is shown. Values represent the ratio of p-p38/total p38 normalized to time-matched vehicle control (A). JC cells were pretreated with the p38 inhibitor SB203580 (“SB”; 10 μM, 30 min), and then 15d-PGJ 2 (0.3 μM) was added for an additional 4 h. Cells were fixed in 3% glutaraldehyde and focal adhesions quantified using interference reflection microscopy. Values represent the mean percentage of cells scored positive for focal adhesions (B). JC cells were also pretreated with SB (10 μM, 30 min), and then BD-15d-PGJ 2 (0.24 μM) was added for an additional 30 min. Cells were then fixed, permeabilized and stained with 2 units of Alexa Fluor® 633 Phalloidin to visualize F-actin (red channel). Nuclei were visualized with DAPI (blue channel). Representative images of red and blue channel merged images are shown from samples prepared in triplicate. EtOH and DMSO were used as vehicle controls. (C). Values shown represent means ± SEM, n = at least 3-6. ** p

    Journal: The Biochemical journal

    Article Title: Modulation of mammary cancer cell migration by 15-deoxy-?12,14-prostaglandin J2: implications for anti-metastatic therapy

    doi: 10.1042/BJ20091193

    Figure Lengend Snippet: Role of p38 in focal adhesion disassembly and cytoskeletal arrangement regulation JC cells were treated with 15d-PGJ 2 (0.3 μM, 15 and 30 min) or ethanol (EtOH) as a vehicle control and phosphorylated p38 (p-p38) was determined by Western blot analysis and quantified. A representative Western blot image is shown. Values represent the ratio of p-p38/total p38 normalized to time-matched vehicle control (A). JC cells were pretreated with the p38 inhibitor SB203580 (“SB”; 10 μM, 30 min), and then 15d-PGJ 2 (0.3 μM) was added for an additional 4 h. Cells were fixed in 3% glutaraldehyde and focal adhesions quantified using interference reflection microscopy. Values represent the mean percentage of cells scored positive for focal adhesions (B). JC cells were also pretreated with SB (10 μM, 30 min), and then BD-15d-PGJ 2 (0.24 μM) was added for an additional 30 min. Cells were then fixed, permeabilized and stained with 2 units of Alexa Fluor® 633 Phalloidin to visualize F-actin (red channel). Nuclei were visualized with DAPI (blue channel). Representative images of red and blue channel merged images are shown from samples prepared in triplicate. EtOH and DMSO were used as vehicle controls. (C). Values shown represent means ± SEM, n = at least 3-6. ** p

    Article Snippet: The p38 inhibitor SB203580 was purchased from Calbiochem (San Diego, CA).

    Techniques: Western Blot, Microscopy, Staining

    Effects of inhibition of MAPK pathways and NF-κB pathways on H. pylori -induced promoter binding in MKN28 cells. Each inhibitor was added to MKN28 cells 30 min before H. pylori infection. Cells were then incubated for 2 h. Nuclear extracts were prepared from control and H. pylori -infected MKN28 cells and used for EMSA. The binding complex of AP-1 was inhibited by each MAPK inhibitor, but not by MG-132. The binding complex of CRE was also inhibited by SB203580 and U0126; however SP600125 and MG-132 had no effect. The binding complex of C/EBP was slightly inhibited by either the MAPK inhibitors or MG-132. The binding complex of NF-κB was inhibited by MG-132; however, it was not inhibited by MAPK inhibitors, suggesting that MAPK inhibitors impeded NF-κB transactivation, but not DNA binding.

    Journal: Molecular Biology of the Cell

    Article Title: Regulation of Interleukin-6 Promoter Activation in Gastric Epithelial Cells Infected with Helicobacter pylori

    doi: 10.1091/mbc.E05-05-0426

    Figure Lengend Snippet: Effects of inhibition of MAPK pathways and NF-κB pathways on H. pylori -induced promoter binding in MKN28 cells. Each inhibitor was added to MKN28 cells 30 min before H. pylori infection. Cells were then incubated for 2 h. Nuclear extracts were prepared from control and H. pylori -infected MKN28 cells and used for EMSA. The binding complex of AP-1 was inhibited by each MAPK inhibitor, but not by MG-132. The binding complex of CRE was also inhibited by SB203580 and U0126; however SP600125 and MG-132 had no effect. The binding complex of C/EBP was slightly inhibited by either the MAPK inhibitors or MG-132. The binding complex of NF-κB was inhibited by MG-132; however, it was not inhibited by MAPK inhibitors, suggesting that MAPK inhibitors impeded NF-κB transactivation, but not DNA binding.

    Article Snippet: In some experiments, cells were pretreated (30 min before H. pylori infection) with the mitogen-activated protein kinase (MAPK) inhibitor (U0126, SB203580, or SP600125) or the proteasome inhibitor, N-cbz-Leu-Leu-leucinal MG-132 (MG-132; Calbiochem, San Diego, CA).

    Techniques: Inhibition, Binding Assay, Infection, Incubation

    TGF-β1- and EGF-mediated induction of COX-2 is inhibited by PD98059 (75 µM), SB203580 (10 µM) and AG1478 (50 µM) in Mv1Lu cells. PD98059, SB203580 and AG1478 compounds were added in the serum-free media 1 hour prior to the addition of TGF-β1 and EGF. After 8 hours of treatment with the growth factors, cells were lysed and 50 µg of each cell lysate was used for immunoblotting.

    Journal: Neoplasia (New York, N.Y.)

    Article Title: Synergistic Induction of Cyclooxygenase-2 by Transforming Growth Factor-β1 and Epidermal Growth Factor Inhibits Apoptosis in Epithelial Cells 1

    doi:

    Figure Lengend Snippet: TGF-β1- and EGF-mediated induction of COX-2 is inhibited by PD98059 (75 µM), SB203580 (10 µM) and AG1478 (50 µM) in Mv1Lu cells. PD98059, SB203580 and AG1478 compounds were added in the serum-free media 1 hour prior to the addition of TGF-β1 and EGF. After 8 hours of treatment with the growth factors, cells were lysed and 50 µg of each cell lysate was used for immunoblotting.

    Article Snippet: Selective kinase inhibitors including PD98059, SB203580 and AG1478 were purchased from Calbiochem.

    Techniques:

    Involvement of p38 MAPK in ethanol induction of ISRE transcription and Stat1 serine phosphorylation. Panel A, Huh7 cells were transfected with 0.7 μg pISRE-luc, and 22 hours later, cells were incubated for 2 hours at the indicated μM concentrations of SB203508 (a p38 inhibitor), followed by 100 mM ethanol. Cell lysates were harvested 6 hours later and luciferase results were normalized to amounts of total cellular protein. Error bars represent standard deviations. The experiment was repeated 4 times with identical results. B, Huh7 cells were treated for 2 hours in the presence of 50 μM of various MAPK inhibitors, and stimulated with 100 mM alcohol for 20 minutes. Whole cell protein extracts were blotted for the serine phosphorylated form (S727) or total form of Stat1. The experiment was repeated twice, yielding similar results. C, Huh7 cells were transfected with control vector plasmid (Vec) or a plasmid expressing a dominant negative mutant p38 protein (p38 AGF). Twenty-four hours later, cells were not treated or treated for 20 minutes with 100 mM ethanol. Levels of S727 and total Stat1 and transfected p38 proteins were determined by western blot. The figure is representative of 2 independent experiments that produced similar results.

    Journal: Virology Journal

    Article Title: Effect of ethanol on innate antiviral pathways and HCV replication in human liver cells

    doi: 10.1186/1743-422X-2-89

    Figure Lengend Snippet: Involvement of p38 MAPK in ethanol induction of ISRE transcription and Stat1 serine phosphorylation. Panel A, Huh7 cells were transfected with 0.7 μg pISRE-luc, and 22 hours later, cells were incubated for 2 hours at the indicated μM concentrations of SB203508 (a p38 inhibitor), followed by 100 mM ethanol. Cell lysates were harvested 6 hours later and luciferase results were normalized to amounts of total cellular protein. Error bars represent standard deviations. The experiment was repeated 4 times with identical results. B, Huh7 cells were treated for 2 hours in the presence of 50 μM of various MAPK inhibitors, and stimulated with 100 mM alcohol for 20 minutes. Whole cell protein extracts were blotted for the serine phosphorylated form (S727) or total form of Stat1. The experiment was repeated twice, yielding similar results. C, Huh7 cells were transfected with control vector plasmid (Vec) or a plasmid expressing a dominant negative mutant p38 protein (p38 AGF). Twenty-four hours later, cells were not treated or treated for 20 minutes with 100 mM ethanol. Levels of S727 and total Stat1 and transfected p38 proteins were determined by western blot. The figure is representative of 2 independent experiments that produced similar results.

    Article Snippet: MAPK inhibitors UO126, PD98059, and SB203508, used to inhibit p42/44, MEK1, and p38 MAPK pathways, respectively, were solubilized in DMSO and obtained from Calbiochem.

    Techniques: Transfection, Incubation, Luciferase, Plasmid Preparation, Expressing, Dominant Negative Mutation, Western Blot, Produced