sp600125 Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore sp600125
    Kinetic inhibition of MAPKs. ELISA quantitation of IL-8 concentrations in cells pretreated with ERK1/2 PD980559, JNK <t>SP600125,</t> and SB203580 inhibitors. Control is HEp-2 cells infected with live HPIV-1. (a) ERK1/2 inhibitor assay is displayed in red cells treated with the inhibitor. (b) JNK inhibitor assay, green, occurs in cells treated with the inhibitor. (c) p38 inhibitor assay, purple, occurs in cells treated with inhibitor. Data are the means ± SD for triplicate measurements from the three separate experiments.
    Sp600125, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 7026 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp600125/product/Millipore
    Average 99 stars, based on 7026 article reviews
    Price from $9.99 to $1999.99
    sp600125 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Tocris sp600125
    Long-term treatment with JNK inhibitor continues to reversibly prevent axon formation. a , Neurons transfected to express soluble tomato fluorescent protein and immunostained for tau-1. Top panels, Representative neuron chronically treated with <t>SP600125</t> for 4 d (4 dy), showing continued inhibition of axon formation. Bottom panels, Representative neuron cultured for 4 d in the chronic presence of SP600125, followed by washout for 24 h. The neuron formed a single axon. b , Neurons at 7 d (7 dy) in culture immunostained for tau-1 and MAP2. Top panels, Representative neurons chronically treated with SP600125. The neurons did not form axons but had MAP2-immunopositive neurites that resembled dendrites. Bottom panels, Representative neurons cultured in the presence of SP600125 for 6 d, followed by washout for 24 h. Two of the four neurons formed axons (arrows). Scale bars: a , 25 μm (top panels) and 50 μm (bottom panels); b , 50 μm. SP, SP600125.
    Sp600125, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 739 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp600125/product/Tocris
    Average 99 stars, based on 739 article reviews
    Price from $9.99 to $1999.99
    sp600125 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Selleck Chemicals sp 600125
    Activation of the MAPK signaling pathway by paramylon. a The levels of p-p38, p38, p-SAPK/JNK, SAPK/JNK, p-ERK, ERK were detected by Western blot analysis and normalized to GAPDH. Lipopolysaccharide (LPS, 100 ng/mL) was used as positive control. RAW264.7 cells in 96-well plates (1 × 10 5 cells/well) were incubated with 200 μg/mL paramylon with, p38-inhibitor SB 20358 (10 μM), JNK inhibitor <t>SP</t> 600125 (10 μM) and ERK inhibitor PD 98059 (10 μM) to measure NO ( b ), TNF-α ( c ) and IL-6 ( d ) levels in the culture medium. Representative results from three independent experiments are shown, * P
    Sp 600125, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sp 600125/product/Selleck Chemicals
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    sp 600125 - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    88
    Avantor 1 9 pyrazoloanthrone
    Activation of the MAPK signaling pathway by paramylon. a The levels of p-p38, p38, p-SAPK/JNK, SAPK/JNK, p-ERK, ERK were detected by Western blot analysis and normalized to GAPDH. Lipopolysaccharide (LPS, 100 ng/mL) was used as positive control. RAW264.7 cells in 96-well plates (1 × 10 5 cells/well) were incubated with 200 μg/mL paramylon with, p38-inhibitor SB 20358 (10 μM), JNK inhibitor <t>SP</t> 600125 (10 μM) and ERK inhibitor PD 98059 (10 μM) to measure NO ( b ), TNF-α ( c ) and IL-6 ( d ) levels in the culture medium. Representative results from three independent experiments are shown, * P
    1 9 Pyrazoloanthrone, supplied by Avantor, 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/1 9 pyrazoloanthrone/product/Avantor
    Average 88 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    1 9 pyrazoloanthrone - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

    Image Search Results


    Kinetic inhibition of MAPKs. ELISA quantitation of IL-8 concentrations in cells pretreated with ERK1/2 PD980559, JNK SP600125, and SB203580 inhibitors. Control is HEp-2 cells infected with live HPIV-1. (a) ERK1/2 inhibitor assay is displayed in red cells treated with the inhibitor. (b) JNK inhibitor assay, green, occurs in cells treated with the inhibitor. (c) p38 inhibitor assay, purple, occurs in cells treated with inhibitor. Data are the means ± SD for triplicate measurements from the three separate experiments.

    Journal: Journal of Immunology Research

    Article Title: Parainfluenza Virus Type 1 Induces Epithelial IL-8 Production via p38-MAPK Signalling

    doi: 10.1155/2014/515984

    Figure Lengend Snippet: Kinetic inhibition of MAPKs. ELISA quantitation of IL-8 concentrations in cells pretreated with ERK1/2 PD980559, JNK SP600125, and SB203580 inhibitors. Control is HEp-2 cells infected with live HPIV-1. (a) ERK1/2 inhibitor assay is displayed in red cells treated with the inhibitor. (b) JNK inhibitor assay, green, occurs in cells treated with the inhibitor. (c) p38 inhibitor assay, purple, occurs in cells treated with inhibitor. Data are the means ± SD for triplicate measurements from the three separate experiments.

    Article Snippet: IL-8 Quantitation in the Presence of Kinase Inhibitors For the best reproduction of larynx conditions, assays were conducted using HEp-2 cells that were pretreated with the kinase inhibitors PD980559 for ERK1/2, SB203580* for p38 MAPK, and SP600125* for JNK from Calbiochem Millipore* (San Diego, CA).

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Infection

    Inhibitors of ERK1/2 PD980559, JNK SP600125, and p38 SB203580 inhibited the expression of each of the MAPKs in the HEp-2 cell line treated cell 1 hour before infection and maintained for 120 minutes after the test. The experiments were performed by adding inhibitor at concentrations from 5 μ M to 20 μ M (1.3 to 5.2 μ L, resp.). Protein levels were detected by Western blot. Immunoblotting was done with monoclonal antibody specific for protein phosphorylated technique that was described in “Materials and Methods”. The extracted proteins (−) represent HEp-2 cells without infecting and the proteins (+) refer to cells infected with the HPIV-1. The tests were performed in triplicate.

    Journal: Journal of Immunology Research

    Article Title: Parainfluenza Virus Type 1 Induces Epithelial IL-8 Production via p38-MAPK Signalling

    doi: 10.1155/2014/515984

    Figure Lengend Snippet: Inhibitors of ERK1/2 PD980559, JNK SP600125, and p38 SB203580 inhibited the expression of each of the MAPKs in the HEp-2 cell line treated cell 1 hour before infection and maintained for 120 minutes after the test. The experiments were performed by adding inhibitor at concentrations from 5 μ M to 20 μ M (1.3 to 5.2 μ L, resp.). Protein levels were detected by Western blot. Immunoblotting was done with monoclonal antibody specific for protein phosphorylated technique that was described in “Materials and Methods”. The extracted proteins (−) represent HEp-2 cells without infecting and the proteins (+) refer to cells infected with the HPIV-1. The tests were performed in triplicate.

    Article Snippet: IL-8 Quantitation in the Presence of Kinase Inhibitors For the best reproduction of larynx conditions, assays were conducted using HEp-2 cells that were pretreated with the kinase inhibitors PD980559 for ERK1/2, SB203580* for p38 MAPK, and SP600125* for JNK from Calbiochem Millipore* (San Diego, CA).

    Techniques: Expressing, Infection, Western Blot

    Effect of inhibition of JNK on HLMVEC network formation in Matrigel. A : representative bright field images of HLMVECs plated on Matrigel as described in MATERIALS AND METHODS in the presence of vehicle (DMSO), SP-600125 (5 μM), or the negative

    Journal:

    Article Title: Role of JNK in network formation of human lung microvascular endothelial cells

    doi: 10.1152/ajplung.00496.2007

    Figure Lengend Snippet: Effect of inhibition of JNK on HLMVEC network formation in Matrigel. A : representative bright field images of HLMVECs plated on Matrigel as described in MATERIALS AND METHODS in the presence of vehicle (DMSO), SP-600125 (5 μM), or the negative

    Article Snippet: The ERK1/2 inhibitors PD 98059, U0126, SB-203580, and SP-600125 and negative JNK control (N1 -Methyl-1,9-pyrazoloanthrone, cat. no. 420123) were purchased from Calbiochem, La Jolla, CA.

    Techniques: Inhibition

    WISP1 inhibits DOX-induced MAPK activation and cardiomyocyte death A , DOX induces p38 MAPK activation. Cardiomyocytes were treated with DOX (1 μM) for the indicated time periods. p38 MAPK activation was analyzed by immunoblotting using activation-specific antibodies (p-p38 MAPK Thr180/Tyr182) and cleared whole cell homogenates (n=3). B , DOX-mediated p38 MAPK activation is inhibited by SB 203580. Cardiomyocytes were treated with SB 203580 (SB; 1 μM in DMSO for 30 min) prior to DOX (1 μM for 30 min) addition, and then analyzed as in A (n=3). C , DOX induces JNK activation. Cardiomyocytes treated as in A were analyzed for JNK activation by immunoblotting using activation-specific antibodies and cleared whole cell homogenates (n=3). D , SP 600125 inhibits DOX-mediated JNK activation (pJNK Thr183/Tyr185). Cardiomyocytes were treated with SP 600125 (SP; 20 μM in DMSO for 30 min) prior to DOX (1 μM for 30 min) treatment, and then analyzed as in C (n=3). E , SB and SP attenuate DOX-induced Bax translocation to mitochondria. Cardiomyocytes were treated with SB and SP prior to DOX addition (1 μM for 8 h). Bax levels in mitochondria were analyzed by immunoblotting. V-β served as a loading and purity control. F , SB and SP attenuate DOX-induced cardiomyocyte death. Cardiomyocytes were treated with SB and SP prior to DOX addition (1 μM for 24 h). Cell death was analyzed as in 1 A . * P

    Journal: Cellular signalling

    Article Title: WNT1-Inducible Signaling Pathway Protein-1 Activates Diverse Cell Survival Pathways and Blocks Doxorubicin-induced Cardiomyocyte Death

    doi: 10.1016/j.cellsig.2010.01.005

    Figure Lengend Snippet: WISP1 inhibits DOX-induced MAPK activation and cardiomyocyte death A , DOX induces p38 MAPK activation. Cardiomyocytes were treated with DOX (1 μM) for the indicated time periods. p38 MAPK activation was analyzed by immunoblotting using activation-specific antibodies (p-p38 MAPK Thr180/Tyr182) and cleared whole cell homogenates (n=3). B , DOX-mediated p38 MAPK activation is inhibited by SB 203580. Cardiomyocytes were treated with SB 203580 (SB; 1 μM in DMSO for 30 min) prior to DOX (1 μM for 30 min) addition, and then analyzed as in A (n=3). C , DOX induces JNK activation. Cardiomyocytes treated as in A were analyzed for JNK activation by immunoblotting using activation-specific antibodies and cleared whole cell homogenates (n=3). D , SP 600125 inhibits DOX-mediated JNK activation (pJNK Thr183/Tyr185). Cardiomyocytes were treated with SP 600125 (SP; 20 μM in DMSO for 30 min) prior to DOX (1 μM for 30 min) treatment, and then analyzed as in C (n=3). E , SB and SP attenuate DOX-induced Bax translocation to mitochondria. Cardiomyocytes were treated with SB and SP prior to DOX addition (1 μM for 8 h). Bax levels in mitochondria were analyzed by immunoblotting. V-β served as a loading and purity control. F , SB and SP attenuate DOX-induced cardiomyocyte death. Cardiomyocytes were treated with SB and SP prior to DOX addition (1 μM for 24 h). Cell death was analyzed as in 1 A . * P

    Article Snippet: PI3K inhibitors (25 μM in DMSO for 1 h) and wortmannin (250 nM in DMSO for 1 h), Akt inhibitor X (Akti-X, # 124040, 2.5 μM in water for 1 h), p38 MAPK inhibitor SB 203580 (1 μM in DMSO for 30 min), JNK inhibitor SP 600125 (20 μM in DMSO for 30 min), proteasomal inhibitor lactacystin (5 μM in DMSO for 30 min), and DMSO were purchased from EMD Biosciences (Gibbstown, NJ 08027).

    Techniques: Activation Assay, Translocation Assay

    Long-term treatment with JNK inhibitor continues to reversibly prevent axon formation. a , Neurons transfected to express soluble tomato fluorescent protein and immunostained for tau-1. Top panels, Representative neuron chronically treated with SP600125 for 4 d (4 dy), showing continued inhibition of axon formation. Bottom panels, Representative neuron cultured for 4 d in the chronic presence of SP600125, followed by washout for 24 h. The neuron formed a single axon. b , Neurons at 7 d (7 dy) in culture immunostained for tau-1 and MAP2. Top panels, Representative neurons chronically treated with SP600125. The neurons did not form axons but had MAP2-immunopositive neurites that resembled dendrites. Bottom panels, Representative neurons cultured in the presence of SP600125 for 6 d, followed by washout for 24 h. Two of the four neurons formed axons (arrows). Scale bars: a , 25 μm (top panels) and 50 μm (bottom panels); b , 50 μm. SP, SP600125.

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: Long-term treatment with JNK inhibitor continues to reversibly prevent axon formation. a , Neurons transfected to express soluble tomato fluorescent protein and immunostained for tau-1. Top panels, Representative neuron chronically treated with SP600125 for 4 d (4 dy), showing continued inhibition of axon formation. Bottom panels, Representative neuron cultured for 4 d in the chronic presence of SP600125, followed by washout for 24 h. The neuron formed a single axon. b , Neurons at 7 d (7 dy) in culture immunostained for tau-1 and MAP2. Top panels, Representative neurons chronically treated with SP600125. The neurons did not form axons but had MAP2-immunopositive neurites that resembled dendrites. Bottom panels, Representative neurons cultured in the presence of SP600125 for 6 d, followed by washout for 24 h. Two of the four neurons formed axons (arrows). Scale bars: a , 25 μm (top panels) and 50 μm (bottom panels); b , 50 μm. SP, SP600125.

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Transfection, Inhibition, Cell Culture

    JNK inhibition results in neurites expressing the dendritic marker, MAP2, but not the axonal marker, tau-1. Control (CTL) neurons at 24 h in culture ( a ) or chronically SP600125 (SP)-treated neurons at 48 h in culture ( b ) revealed that SP600125-treated neurons had MAP2-positive, but tau-1-negative, neurites. Arrows denote axons. Scale bars, 25 μm.

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: JNK inhibition results in neurites expressing the dendritic marker, MAP2, but not the axonal marker, tau-1. Control (CTL) neurons at 24 h in culture ( a ) or chronically SP600125 (SP)-treated neurons at 48 h in culture ( b ) revealed that SP600125-treated neurons had MAP2-positive, but tau-1-negative, neurites. Arrows denote axons. Scale bars, 25 μm.

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Inhibition, Expressing, Marker, CTL Assay

    JNK inhibition does not affect the development of minor processes. Average neurite lengths at 48 h in culture under control conditions (open bars; n = 63 neurons, 350 neurites) or with SP600125 treatment (filled bars; n = 108 neurons, 752 neurites) are shown. ∗∗∗ p

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: JNK inhibition does not affect the development of minor processes. Average neurite lengths at 48 h in culture under control conditions (open bars; n = 63 neurons, 350 neurites) or with SP600125 treatment (filled bars; n = 108 neurons, 752 neurites) are shown. ∗∗∗ p

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Inhibition

    Axon formation was reversibly blocked by JNK inhibition. a , b , Control (CTL) neurons at 24 and 48 h in culture. c , d , Neurons chronically treated with the JNK inhibitor SP600125. e , Neurons incubated with SP600125 for 48 h, followed by washout for 24 h. f , Quantification of axon-bearing neurons under control conditions (white bars), after chronic SP600125 treatment (black bars), or with SP600125 washout (gray bar). Axons from individual cells could not be accurately quantified in 72 h control cultures, because the cultures had formed too dense of a network. Error bars indicate SEM. ∗∗∗∗ p

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: Axon formation was reversibly blocked by JNK inhibition. a , b , Control (CTL) neurons at 24 and 48 h in culture. c , d , Neurons chronically treated with the JNK inhibitor SP600125. e , Neurons incubated with SP600125 for 48 h, followed by washout for 24 h. f , Quantification of axon-bearing neurons under control conditions (white bars), after chronic SP600125 treatment (black bars), or with SP600125 washout (gray bar). Axons from individual cells could not be accurately quantified in 72 h control cultures, because the cultures had formed too dense of a network. Error bars indicate SEM. ∗∗∗∗ p

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Inhibition, CTL Assay, Incubation

    Phosphorylation of the JNK substrate ATF-2 is attenuated by SP600125 (SP). a , Western blots demonstrating that ATF-2 was dually phosphorylated (at T51/T53) under basal conditions, which decreased with acute or chronic inhibition of JNK. CTL, Control. b , Quantification revealed JNK inhibition significantly decreased ATF-2 phosphorylation ( n = 6 for each condition). ∗∗∗ p

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: Phosphorylation of the JNK substrate ATF-2 is attenuated by SP600125 (SP). a , Western blots demonstrating that ATF-2 was dually phosphorylated (at T51/T53) under basal conditions, which decreased with acute or chronic inhibition of JNK. CTL, Control. b , Quantification revealed JNK inhibition significantly decreased ATF-2 phosphorylation ( n = 6 for each condition). ∗∗∗ p

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Western Blot, Inhibition, CTL Assay

    Phospho-JNK levels dramatically decreased with SP600125 treatment. a , Representative Western blots of cultures 24 h after plating that were chronically treated with SP600125 (SP 24Hr), treated with SP600125 for 90 min just before lysing (SP 90 min), or untreated (control). b , Graph showing that chronic and acute treatment with SP600125 significantly inhibited phosphorylation of JNK but did not affect total JNK levels ( n = 6 for each condition). ∗∗∗ p

    Journal: The Journal of Neuroscience

    Article Title: Activated c-Jun N-Terminal Kinase Is Required for Axon Formation

    doi: 10.1523/JNEUROSCI.2625-06.2006

    Figure Lengend Snippet: Phospho-JNK levels dramatically decreased with SP600125 treatment. a , Representative Western blots of cultures 24 h after plating that were chronically treated with SP600125 (SP 24Hr), treated with SP600125 for 90 min just before lysing (SP 90 min), or untreated (control). b , Graph showing that chronic and acute treatment with SP600125 significantly inhibited phosphorylation of JNK but did not affect total JNK levels ( n = 6 for each condition). ∗∗∗ p

    Article Snippet: The neurites that formed in the presence of SP600125 resembled dendrites in their branching and taper.

    Techniques: Western Blot

    ENT 1/2 and MAK kinases mediated sustained adenosine-induced mitochondrial oxidative stress. PAECs were pre-treated with ENT 1/2 inhibitors, DPM (10 µM) or NBTI (10 µM), an A 2A R antagonist, DPMX (10 µM), or an A 2B R antagonist, MRS1754 (10 µM), for 1 h and then incubated with 50 µM adenosine and 50 µM DCF (AD) in the absence or presence of individual inhibitor for 30 h (Panel a). Similarly, the cells were pre-treated with a cytosolic antioxidant, NAC (12.5 mM), a mitochondrial ROS scavenger, mitoTEMPO (5 µM), a p38 inhibitor, SB203580 (SB, 10 µM), or a JNK inhibitor, SP600125 (SP, 10 µM), and then incubated with 50 µM adenosine and 50 µM DCF (AD) in the absence or presence of individual inhibitors for 30 h (Panel b). Following the treatments, the cells were washed twice and incubated with Hoechst 33342 and MitoSOX Red to quantify cell numbers and mitochondrial ROS levels. The relative numbers of cells in each well were normalized by the fluorescence intensity of Hoechst 33342. The fluorescence intensity of MitoSOX Red in each well was then adjusted by the relative numbers of cells. The levels of mitochondrial ROS were expressed as fold changes of the fluorescence intensity of MitoSOX Red relative to that of untreated cells. The data are presented as mean ± SD of three independent experiments ( n = 3). * p

    Journal: Pulmonary Circulation

    Article Title: Sustained adenosine exposure causes endothelial mitochondrial dysfunction via equilibrative nucleoside transporters

    doi: 10.1177/2045894020924994

    Figure Lengend Snippet: ENT 1/2 and MAK kinases mediated sustained adenosine-induced mitochondrial oxidative stress. PAECs were pre-treated with ENT 1/2 inhibitors, DPM (10 µM) or NBTI (10 µM), an A 2A R antagonist, DPMX (10 µM), or an A 2B R antagonist, MRS1754 (10 µM), for 1 h and then incubated with 50 µM adenosine and 50 µM DCF (AD) in the absence or presence of individual inhibitor for 30 h (Panel a). Similarly, the cells were pre-treated with a cytosolic antioxidant, NAC (12.5 mM), a mitochondrial ROS scavenger, mitoTEMPO (5 µM), a p38 inhibitor, SB203580 (SB, 10 µM), or a JNK inhibitor, SP600125 (SP, 10 µM), and then incubated with 50 µM adenosine and 50 µM DCF (AD) in the absence or presence of individual inhibitors for 30 h (Panel b). Following the treatments, the cells were washed twice and incubated with Hoechst 33342 and MitoSOX Red to quantify cell numbers and mitochondrial ROS levels. The relative numbers of cells in each well were normalized by the fluorescence intensity of Hoechst 33342. The fluorescence intensity of MitoSOX Red in each well was then adjusted by the relative numbers of cells. The levels of mitochondrial ROS were expressed as fold changes of the fluorescence intensity of MitoSOX Red relative to that of untreated cells. The data are presented as mean ± SD of three independent experiments ( n = 3). * p

    Article Snippet: 2′-Deoxycoformycin (DCF) and SP600125 (SP) were purchased from Tocris Bioscience (Minneapolis, MN).

    Techniques: Incubation, Fluorescence

    Effect of MEK and ERK on ERRγ protein levels A, Inhibition of ERK, but not p38 or JNK, reduces exogenous ERRγ expression. MCF7 cells were transiently transfected with the pSG5 empty vector or HA-ERRγ, then treated with DMSO vehicle, 5 μM U0126 (MEK inhibitor), 25 μM SB203580 (p38 inhibitor), or 10 μM SP600125 (JNK inhibitor) for 24 hours prior to lysis and Western blot analysis. Left panels show ERRγ (HA) levels, phosphorylated ERK (pERK), and total ERK from a representative experiment repeated at least twice. Right panels show total and phosphorylated p38 and JNK (p-p38 and pJNK, respectively) from the same experiment. β–actin = loading control. B, Constitutively active, mutant MEK enhances ERRγ protein levels. MCF7 cells were transiently co-transfected with HA-ERRγ and either MEKDD or additional pSG5 empty vector. *denotes the transfected MEKDD construct. β–actin = loading control. C, Exogenous, wild type ERK2 enhances ERRγ protein levels. MCF7 cells were transiently co-transfected with HA-ERRγ and either MEKDD, wild type HA-tagged ERK2, or additional pSG5 empty vector. *denotes the transfected MEKDD construct. The arrowhead and ^ denote transfected HA-ERRγ and HA-ERK2, respectively. β–actin = loading control. D, EGF-mediated enhancement of ERRγ protein levels is reversed by concomitant ERK inhibition. MCF7 cells were transiently transfected with HA-ERRγ, then cultured in low-serum conditions for 20 hours before treatment with DMSO vehicle, 25 ng/ml EGF, or 25 ng/ml EGF plus 5 μM U0126 for 2 hours. β–actin = loading control. E, Inhibition of ERK reduces exogenous ERRγ expression in a second ER+ breast cancer cell line. SUM44 cells were transiently transfected with the pSG5 empty vector or HA-ERRγ, then treated with DMSO vehicle or 5 μM U0126 for 22 hours. β–actin = loading control.

    Journal: The FEBS journal

    Article Title: ERK/MAPK regulates ERRγ expression, transcriptional activity, and receptor-mediated Tamoxifen resistance in ER+ breast cancer

    doi: 10.1111/febs.12797

    Figure Lengend Snippet: Effect of MEK and ERK on ERRγ protein levels A, Inhibition of ERK, but not p38 or JNK, reduces exogenous ERRγ expression. MCF7 cells were transiently transfected with the pSG5 empty vector or HA-ERRγ, then treated with DMSO vehicle, 5 μM U0126 (MEK inhibitor), 25 μM SB203580 (p38 inhibitor), or 10 μM SP600125 (JNK inhibitor) for 24 hours prior to lysis and Western blot analysis. Left panels show ERRγ (HA) levels, phosphorylated ERK (pERK), and total ERK from a representative experiment repeated at least twice. Right panels show total and phosphorylated p38 and JNK (p-p38 and pJNK, respectively) from the same experiment. β–actin = loading control. B, Constitutively active, mutant MEK enhances ERRγ protein levels. MCF7 cells were transiently co-transfected with HA-ERRγ and either MEKDD or additional pSG5 empty vector. *denotes the transfected MEKDD construct. β–actin = loading control. C, Exogenous, wild type ERK2 enhances ERRγ protein levels. MCF7 cells were transiently co-transfected with HA-ERRγ and either MEKDD, wild type HA-tagged ERK2, or additional pSG5 empty vector. *denotes the transfected MEKDD construct. The arrowhead and ^ denote transfected HA-ERRγ and HA-ERK2, respectively. β–actin = loading control. D, EGF-mediated enhancement of ERRγ protein levels is reversed by concomitant ERK inhibition. MCF7 cells were transiently transfected with HA-ERRγ, then cultured in low-serum conditions for 20 hours before treatment with DMSO vehicle, 25 ng/ml EGF, or 25 ng/ml EGF plus 5 μM U0126 for 2 hours. β–actin = loading control. E, Inhibition of ERK reduces exogenous ERRγ expression in a second ER+ breast cancer cell line. SUM44 cells were transiently transfected with the pSG5 empty vector or HA-ERRγ, then treated with DMSO vehicle or 5 μM U0126 for 22 hours. β–actin = loading control.

    Article Snippet: The MEK inhibitor U0126, JNK inhibitor SP600125 and p38 inhibitor SB203580 (Tocris Bioscience, Ellisville, MO) were dissolved in dimethyl sulfoxide (DMSO), stored as 10 and 50mM stocks (respectively) at −20°C, and used at the concentrations indicated.

    Techniques: Inhibition, Expressing, Transfection, Plasmid Preparation, Lysis, Western Blot, Mutagenesis, Construct, Cell Culture

    Activation of the MAPK signaling pathway by paramylon. a The levels of p-p38, p38, p-SAPK/JNK, SAPK/JNK, p-ERK, ERK were detected by Western blot analysis and normalized to GAPDH. Lipopolysaccharide (LPS, 100 ng/mL) was used as positive control. RAW264.7 cells in 96-well plates (1 × 10 5 cells/well) were incubated with 200 μg/mL paramylon with, p38-inhibitor SB 20358 (10 μM), JNK inhibitor SP 600125 (10 μM) and ERK inhibitor PD 98059 (10 μM) to measure NO ( b ), TNF-α ( c ) and IL-6 ( d ) levels in the culture medium. Representative results from three independent experiments are shown, * P

    Journal: BMC Microbiology

    Article Title: Immune activation of murine RAW264.7 macrophages by sonicated and alkalized paramylon from Euglena gracilis

    doi: 10.1186/s12866-020-01782-y

    Figure Lengend Snippet: Activation of the MAPK signaling pathway by paramylon. a The levels of p-p38, p38, p-SAPK/JNK, SAPK/JNK, p-ERK, ERK were detected by Western blot analysis and normalized to GAPDH. Lipopolysaccharide (LPS, 100 ng/mL) was used as positive control. RAW264.7 cells in 96-well plates (1 × 10 5 cells/well) were incubated with 200 μg/mL paramylon with, p38-inhibitor SB 20358 (10 μM), JNK inhibitor SP 600125 (10 μM) and ERK inhibitor PD 98059 (10 μM) to measure NO ( b ), TNF-α ( c ) and IL-6 ( d ) levels in the culture medium. Representative results from three independent experiments are shown, * P

    Article Snippet: The inhibitors: SB 20358, SP 600125 and PD 98059, were purchased from Selleck (Shanghai, China).

    Techniques: Activation Assay, Western Blot, Positive Control, Incubation