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  • 99
    Tocris pd98059
    The ERK and p38 MAPK pathways are involved in the regulation of salinomycin-induced autophagy in prostate cancer cells. ( A , B ) ERK and p38 MAPK protein expression. ( A ) PC-3 cells. ( B ) LNCaP cells. Cells were treated with 2 µM salinomycin for the indicated times. After treatment, total cell lysates were subjected to SDS-PAGE for Western blot analysis; ( C ) the effect of <t>PD98059</t> on salinomycin-induced autophagy; ( D ) the effect of SB203580 on salinomycin-induced autophagy. Cells were pretreated with 10 µM PD98059 or 10 µM SB203580 for 1 h, and acridine orange (AO) stained cells were evaluated by flow cytometry. All data were representative of at least three independent experiments. Data were shown as the mean ± SD. * p
    Pd98059, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 478 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam pd98059
    Inhibition of p38 weakens heat stress-induced phosphorylation of MK5 and MK2. F98 cells underwent pretreatment with or without the indicated inhibitors for 30 min prior to exposure to heat stress or control heat treatment. After the cells received further incubation for 12 h at 37°C, antibodies specific for p-MK2, p-MK5, MK2, MK5 were used to assess the protein levels in the cell lysates. HS, heat stress; p-, phosphorylated; t-, total; MK, mitogen-activated protein kinase-activated protein kinase; SB203580, p38 inhibitor; <t>PD98059,</t> ERK inhibitor; SP600125, JNK inhibitor.
    Pd98059, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 74 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc pd 98059
    Inhibition of p38 weakens heat stress-induced phosphorylation of MK5 and MK2. F98 cells underwent pretreatment with or without the indicated inhibitors for 30 min prior to exposure to heat stress or control heat treatment. After the cells received further incubation for 12 h at 37°C, antibodies specific for p-MK2, p-MK5, MK2, MK5 were used to assess the protein levels in the cell lysates. HS, heat stress; p-, phosphorylated; t-, total; MK, mitogen-activated protein kinase-activated protein kinase; SB203580, p38 inhibitor; <t>PD98059,</t> ERK inhibitor; SP600125, JNK inhibitor.
    Pd 98059, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Selleck Chemicals pd 98059
    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 <t>PD</t> 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
    Pd 98059, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioVision pd98059
    sCLU regulates the cisplatin (DDP)-induced cytotoxicity in the osteosarcoma (OS) cells. (A) The KH OS or sCLU stably transfected KH OS cells (KH OS/sCLU) were treated with 0.01 to 10 μM/ mL DPP for 72 hours in the presence or absence of 10 mM <t>PD98059.</t> Cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results were obtained from three independent experiments. Versus KHOS/sCLU, * P
    Pd98059, supplied by BioVision, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The ERK and p38 MAPK pathways are involved in the regulation of salinomycin-induced autophagy in prostate cancer cells. ( A , B ) ERK and p38 MAPK protein expression. ( A ) PC-3 cells. ( B ) LNCaP cells. Cells were treated with 2 µM salinomycin for the indicated times. After treatment, total cell lysates were subjected to SDS-PAGE for Western blot analysis; ( C ) the effect of PD98059 on salinomycin-induced autophagy; ( D ) the effect of SB203580 on salinomycin-induced autophagy. Cells were pretreated with 10 µM PD98059 or 10 µM SB203580 for 1 h, and acridine orange (AO) stained cells were evaluated by flow cytometry. All data were representative of at least three independent experiments. Data were shown as the mean ± SD. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Inhibition of Autophagy Promotes Salinomycin-Induced Apoptosis via Reactive Oxygen Species-Mediated PI3K/AKT/mTOR and ERK/p38 MAPK-Dependent Signaling in Human Prostate Cancer Cells

    doi: 10.3390/ijms18051088

    Figure Lengend Snippet: The ERK and p38 MAPK pathways are involved in the regulation of salinomycin-induced autophagy in prostate cancer cells. ( A , B ) ERK and p38 MAPK protein expression. ( A ) PC-3 cells. ( B ) LNCaP cells. Cells were treated with 2 µM salinomycin for the indicated times. After treatment, total cell lysates were subjected to SDS-PAGE for Western blot analysis; ( C ) the effect of PD98059 on salinomycin-induced autophagy; ( D ) the effect of SB203580 on salinomycin-induced autophagy. Cells were pretreated with 10 µM PD98059 or 10 µM SB203580 for 1 h, and acridine orange (AO) stained cells were evaluated by flow cytometry. All data were representative of at least three independent experiments. Data were shown as the mean ± SD. * p

    Article Snippet: LY294002, PD98059 and SB203580 were purchased from TOCRIS (Bristol, UK).

    Techniques: Expressing, SDS Page, Western Blot, Staining, Flow Cytometry, Cytometry

    Influence of kinase inhibitors on ppNOC and NOP mRNA expression. ppNOC and NOP expression in blood leukocytes cultured with PMA or without (control) and with or without pre-treatment with the specific kinase inhibitors PD98059 (PD, ERK inhibitor) 30 μM, SB203580 (SB, p38 inhibitor) 10 μM, SP600125 (SP, JNK inhibitor) 10 μM, Bay 11-7821 (Bay, NFκB inhibitor) 3 μM, or the combination of PD and SB. Box-and-whisker plots, medians with interquartile ranges, and 5 to 95 percentiles, n =10 for the PD+SB+PMA group, n =17 for all other groups. * P

    Journal: Molecular Pain

    Article Title: ERK and p38 contribute to the regulation of nociceptin and the nociceptin receptor in human peripheral blood leukocytes

    doi: 10.1177/1744806919828921

    Figure Lengend Snippet: Influence of kinase inhibitors on ppNOC and NOP mRNA expression. ppNOC and NOP expression in blood leukocytes cultured with PMA or without (control) and with or without pre-treatment with the specific kinase inhibitors PD98059 (PD, ERK inhibitor) 30 μM, SB203580 (SB, p38 inhibitor) 10 μM, SP600125 (SP, JNK inhibitor) 10 μM, Bay 11-7821 (Bay, NFκB inhibitor) 3 μM, or the combination of PD and SB. Box-and-whisker plots, medians with interquartile ranges, and 5 to 95 percentiles, n =10 for the PD+SB+PMA group, n =17 for all other groups. * P

    Article Snippet: Blood was pre-treated with PD98059 (PD) 30 μM, SP600125 (SP) 10 μM, SB203580 (SB) 10 μM, Bay 11-7871 (Bay) 3 μM, or the combination of PD and SB (all from Tocris Bioscience, Bristol, UK) for 1 h prior to culturing with or without PMA 10 ng/ml for 6 and 24 h. The concentrations of the inhibitors were based on doses used in previous studies., , A culture without any stimulus and one cultured only with PMA 10 ng/ml served as control group and reference group, respectively.

    Techniques: Expressing, Cell Culture, Whisker Assay

    STC-1 promoted VEGF expressing through PKCβII signaling pathway . (A) VEGF expression in the different culture supernatants. ELISA assay was used to detect VEGF expression in the culture supernatants. (B) Time courses of PKCβIIand ERK1/2 avtivation induced by STC-1. BGC823 were treated with 50 ng/mL STC-1 for 15, 30, 45, 60 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosphor-PKC βII, total PKC βII, phosho-ERK1/2 and total ERK1/2. (C) Concentration courses of PKC βIIand P38 activation induced by STC-1. BGC823 were treated with different concentrations STC-1 for 45 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosphor-PKC βII, total PKC βII, phosho-P38 and total P38. The results are representative of three independent experiments. (D) Concentration courses of ERK1/2 activation induced by STC-1. BGC823 were treated with different concentrations STC-1 for 45 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosho-ERK1/2 and total ERK1/2. The results are representative of three independent experiments. (E) Effect of STC-1 on VEGF is mediated through PKCβII and ERK1/2 signaling. BGC823 was exposed to either CGP53353 (0.5 μM) or PD98059 (25 μM) for three hours and then individually with STC-1 for 24 h. The results are representative of three independent experiments. VEGF expression in BGC823 cell culture supernatants was determined by ELISA. (F) CGP53353 and PD98059 could inhibit PKC βIIand ERK activation, respectively.

    Journal: Journal of Biomedical Science

    Article Title: Stanniocalcin-1 promotes tumor angiogenesis through up-regulation of VEGF in gastric cancer cells

    doi: 10.1186/1423-0127-18-39

    Figure Lengend Snippet: STC-1 promoted VEGF expressing through PKCβII signaling pathway . (A) VEGF expression in the different culture supernatants. ELISA assay was used to detect VEGF expression in the culture supernatants. (B) Time courses of PKCβIIand ERK1/2 avtivation induced by STC-1. BGC823 were treated with 50 ng/mL STC-1 for 15, 30, 45, 60 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosphor-PKC βII, total PKC βII, phosho-ERK1/2 and total ERK1/2. (C) Concentration courses of PKC βIIand P38 activation induced by STC-1. BGC823 were treated with different concentrations STC-1 for 45 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosphor-PKC βII, total PKC βII, phosho-P38 and total P38. The results are representative of three independent experiments. (D) Concentration courses of ERK1/2 activation induced by STC-1. BGC823 were treated with different concentrations STC-1 for 45 min. Whole- cell lysates were prepared and immunoblotted with antibodies to phosho-ERK1/2 and total ERK1/2. The results are representative of three independent experiments. (E) Effect of STC-1 on VEGF is mediated through PKCβII and ERK1/2 signaling. BGC823 was exposed to either CGP53353 (0.5 μM) or PD98059 (25 μM) for three hours and then individually with STC-1 for 24 h. The results are representative of three independent experiments. VEGF expression in BGC823 cell culture supernatants was determined by ELISA. (F) CGP53353 and PD98059 could inhibit PKC βIIand ERK activation, respectively.

    Article Snippet: Material PD98059 (selective inhibitor of ERK signaling pathway) and CGP53353 (selective inhibitor of PKCβII signaling pathway) were obtained from TOCRIS Bioscience Company (Bristol, UK) and Beyotime Institute of Biotechnology (Haimen, China), respectively.

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Activation Assay, Cell Culture

    Effects of the MAPK kinase inhibitor PD98059 on the neuroprotection provided by GK-2 or NGF against H 2 O 2 -induced oxidative stress in HT-22 cells. Hippocampal cells, HT-22, were pre-incubated with LY294002 (100 μM) for 30 min before treatment with GK-2(10 -5 M and 10 -8 M) or NGF (10 -9 M). Cell survival after a 30-min exposure to H 2 O 2 was measured by assessment of MTT metabolism. Data are presented as means±SD. * p

    Journal: Journal of Biomedical Science

    Article Title: Dimeric dipeptide mimetics of the nerve growth factor Loop 4 and Loop 1 activate TRKA with different patterns of intracellular signal transduction

    doi: 10.1186/s12929-015-0198-z

    Figure Lengend Snippet: Effects of the MAPK kinase inhibitor PD98059 on the neuroprotection provided by GK-2 or NGF against H 2 O 2 -induced oxidative stress in HT-22 cells. Hippocampal cells, HT-22, were pre-incubated with LY294002 (100 μM) for 30 min before treatment with GK-2(10 -5 M and 10 -8 M) or NGF (10 -9 M). Cell survival after a 30-min exposure to H 2 O 2 was measured by assessment of MTT metabolism. Data are presented as means±SD. * p

    Article Snippet: Inhibitors of PI3K (LY294002) and MAPK (PD98059) were purchased from Tocris Bioscience (Bristol, UK).

    Techniques: Incubation, MTT Assay

    Aspirin enhances ERK but inhibits RhoA activities . (A) Western blot analysis of MBP, phospho-ERK1/2, and total-ERK1/2 protein levels from cultured OPCs treated with ethanol, 10 μM aspirin, or 10 μM aspirin plus ERK1/2 inhibitor PD98059. Note that aspirin can enhance ERK activity as revealed by phospho-ERK1/2 protein level. GAPDH was used as an internal control. (B) Quantitation of MBP, and phospho-ERK1/2 levels in each group, * p

    Journal: Frontiers in Aging Neuroscience

    Article Title: Aspirin Promotes Oligodendrocyte Precursor Cell Proliferation and Differentiation after White Matter Lesion

    doi: 10.3389/fnagi.2014.00007

    Figure Lengend Snippet: Aspirin enhances ERK but inhibits RhoA activities . (A) Western blot analysis of MBP, phospho-ERK1/2, and total-ERK1/2 protein levels from cultured OPCs treated with ethanol, 10 μM aspirin, or 10 μM aspirin plus ERK1/2 inhibitor PD98059. Note that aspirin can enhance ERK activity as revealed by phospho-ERK1/2 protein level. GAPDH was used as an internal control. (B) Quantitation of MBP, and phospho-ERK1/2 levels in each group, * p

    Article Snippet: However, addition of ERK inhibitor PD98059 (specific ERK kinase inhibitor; Tocris Cookson) either reversed the upregulated MBP expression and the enhanced ERK1/2 activity (Figures A,B).

    Techniques: Western Blot, Cell Culture, Activity Assay, Quantitation Assay

    The effects of ERK and JNK inhibitors on SARS-CoV-induced activation of MAPK. A549 cells were pretreated with MEK inhibitor PD98059 (20 μM; PD) or JNK inhibitor SP600125 (30 μM; SP) for 1 h before incubation with SARS-CoV VLPs (A) or with the viral spike protein (B) for 2 h. Cell lysates were prepared and subjected to Western blot analysis with antibodies against phospho-ERK1/2 (p-ERK), ERK1/2 (ERK), phospho-JNK (p-JNK), and JNK as indicated.

    Journal: Journal of Virology

    Article Title: Upregulation of the Chemokine (C-C Motif) Ligand 2 via a Severe Acute Respiratory Syndrome Coronavirus Spike-ACE2 Signaling Pathway ▿

    doi: 10.1128/JVI.02560-09

    Figure Lengend Snippet: The effects of ERK and JNK inhibitors on SARS-CoV-induced activation of MAPK. A549 cells were pretreated with MEK inhibitor PD98059 (20 μM; PD) or JNK inhibitor SP600125 (30 μM; SP) for 1 h before incubation with SARS-CoV VLPs (A) or with the viral spike protein (B) for 2 h. Cell lysates were prepared and subjected to Western blot analysis with antibodies against phospho-ERK1/2 (p-ERK), ERK1/2 (ERK), phospho-JNK (p-JNK), and JNK as indicated.

    Article Snippet: MEK inhibitor PD98059 and Raf inhibitor GW5074 were purchased from Tocris.

    Techniques: Activation Assay, Incubation, Western Blot

    Phosphorylated and total protein expression levels of p38 MAPK and ERK1/2 in MDA-MB-231 cell line treated with specific inhibitors for these MAPKs and stimulated with TGF-β1 for different periods of time . (A) MDA-MB-231 cells were pre-treated with 20 μM of PD98059 (specific ERK1/2 pharmacological inhibitor) for 1 h and then stimulated with 10 ng/mL TGF-β1 for different periods of time (0, 10 min and 3 h). Total protein lysates from these samples were used to analyze the protein expression levels of total and phosphorylated forms of p38 MAPK by Western blotting. (B) The phosphorylated and total protein expression levels of ERK 1/2 were measured by Western blotting in MDA-MB-231 cells pre-treated with 20 μM of SB203680 (p38 MAPK pharmacological specific inhibitor) for 1 h and stimulated with 10 ng/mL TGF-β1 for 0, 30 min and 1 h. These results were analyzed and used to calculate the p-p38 MAPK/total p38 MAPK and p-ERK1/2/total ERK1/2 ratios, respectively. Results are presented in graphics as mean ± standard errors from three independent experiments. The Western blotting figures are representative of one experiment. *, p

    Journal: BMC Cancer

    Article Title: TGF-?1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

    doi: 10.1186/1471-2407-12-26

    Figure Lengend Snippet: Phosphorylated and total protein expression levels of p38 MAPK and ERK1/2 in MDA-MB-231 cell line treated with specific inhibitors for these MAPKs and stimulated with TGF-β1 for different periods of time . (A) MDA-MB-231 cells were pre-treated with 20 μM of PD98059 (specific ERK1/2 pharmacological inhibitor) for 1 h and then stimulated with 10 ng/mL TGF-β1 for different periods of time (0, 10 min and 3 h). Total protein lysates from these samples were used to analyze the protein expression levels of total and phosphorylated forms of p38 MAPK by Western blotting. (B) The phosphorylated and total protein expression levels of ERK 1/2 were measured by Western blotting in MDA-MB-231 cells pre-treated with 20 μM of SB203680 (p38 MAPK pharmacological specific inhibitor) for 1 h and stimulated with 10 ng/mL TGF-β1 for 0, 30 min and 1 h. These results were analyzed and used to calculate the p-p38 MAPK/total p38 MAPK and p-ERK1/2/total ERK1/2 ratios, respectively. Results are presented in graphics as mean ± standard errors from three independent experiments. The Western blotting figures are representative of one experiment. *, p

    Article Snippet: The pharmacological inhibitors against p38 MAPK (SB203680) and ERK1/2 (PD98059) were obtained from Tocris Bioscience (Bristol, UK).

    Techniques: Expressing, Multiple Displacement Amplification, Western Blot

    In vitro migration (A) and invasion (B) capacities of the MDA-MB-231 cell line upon treatment with TGF-β1 and inhibitors of ERK1/2, p38 MAPK and MMPs . MDA-MB-231 cells were pre-treated for 1 h with 20 μM of either PD98059 or SB203680 (ERK1/2 and p38 MAPK pharmacological inhibitor, respectively) or with 40 μM of GM6001 (a broad-spectrum MMPs inhibitor). After the inhibition treatment these cells were stimulated with 10 ng/mL TGF-β1 and allowed to migrate through uncoated transwells for 8 h (A) or invade through matrigel-coated transwells for 24 h (B). The number of cells at the bottom of the transwell filters was counted at the end of each assay. Results are presented as means ± standard errors from three independent experiments, performed in duplicate, all versus control (cells treated with DMSO, vehicle).

    Journal: BMC Cancer

    Article Title: TGF-?1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

    doi: 10.1186/1471-2407-12-26

    Figure Lengend Snippet: In vitro migration (A) and invasion (B) capacities of the MDA-MB-231 cell line upon treatment with TGF-β1 and inhibitors of ERK1/2, p38 MAPK and MMPs . MDA-MB-231 cells were pre-treated for 1 h with 20 μM of either PD98059 or SB203680 (ERK1/2 and p38 MAPK pharmacological inhibitor, respectively) or with 40 μM of GM6001 (a broad-spectrum MMPs inhibitor). After the inhibition treatment these cells were stimulated with 10 ng/mL TGF-β1 and allowed to migrate through uncoated transwells for 8 h (A) or invade through matrigel-coated transwells for 24 h (B). The number of cells at the bottom of the transwell filters was counted at the end of each assay. Results are presented as means ± standard errors from three independent experiments, performed in duplicate, all versus control (cells treated with DMSO, vehicle).

    Article Snippet: The pharmacological inhibitors against p38 MAPK (SB203680) and ERK1/2 (PD98059) were obtained from Tocris Bioscience (Bristol, UK).

    Techniques: In Vitro, Migration, Multiple Displacement Amplification, Inhibition

    Relative mRNA and protein expression of MMPs and MMPs inhibitors in MDA-MB-231 cells treated with TGF-β1 and an ERK1/2 inhibitor . MDA-MB-231 cells were pre-treated for 1 h with different concentrations (0, 5, 10 or 20 μM) of PD98059 (ERK1/2 pharmacological inhibitor) and then stimulated with 10 ng/mL TGF-β1 by 20 h. Total RNA from these samples was used to analyze the mRNA expression levels of MMPs (MMP-2 and MMP-9) and MMPs inhibitors (TIMP-2 and RECK) by qRT-PCR. The levels of pro-enzyme and active MMP-2 and MMP-9 proteins were evaluated by zymography. Total protein lysates were used to measure the protein expression levels of RECK by Western blotting. The GAPDH protein expression was used as the loading control in Western blotting assays. Conditioned medium from these cultures were also utilized to analyze the TIMP-2 protein levels by Western blotting. Results are presented in graphics as means ± standard errors from three independent experiments. The Western blotting figures are representative of one experiment. For mRNA expression levels fold change: MMP-2 (a versus c: p

    Journal: BMC Cancer

    Article Title: TGF-?1 modulates the homeostasis between MMPs and MMP inhibitors through p38 MAPK and ERK1/2 in highly invasive breast cancer cells

    doi: 10.1186/1471-2407-12-26

    Figure Lengend Snippet: Relative mRNA and protein expression of MMPs and MMPs inhibitors in MDA-MB-231 cells treated with TGF-β1 and an ERK1/2 inhibitor . MDA-MB-231 cells were pre-treated for 1 h with different concentrations (0, 5, 10 or 20 μM) of PD98059 (ERK1/2 pharmacological inhibitor) and then stimulated with 10 ng/mL TGF-β1 by 20 h. Total RNA from these samples was used to analyze the mRNA expression levels of MMPs (MMP-2 and MMP-9) and MMPs inhibitors (TIMP-2 and RECK) by qRT-PCR. The levels of pro-enzyme and active MMP-2 and MMP-9 proteins were evaluated by zymography. Total protein lysates were used to measure the protein expression levels of RECK by Western blotting. The GAPDH protein expression was used as the loading control in Western blotting assays. Conditioned medium from these cultures were also utilized to analyze the TIMP-2 protein levels by Western blotting. Results are presented in graphics as means ± standard errors from three independent experiments. The Western blotting figures are representative of one experiment. For mRNA expression levels fold change: MMP-2 (a versus c: p

    Article Snippet: The pharmacological inhibitors against p38 MAPK (SB203680) and ERK1/2 (PD98059) were obtained from Tocris Bioscience (Bristol, UK).

    Techniques: Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Zymography, Western Blot

    Inhibition of p38 weakens heat stress-induced phosphorylation of MK5 and MK2. F98 cells underwent pretreatment with or without the indicated inhibitors for 30 min prior to exposure to heat stress or control heat treatment. After the cells received further incubation for 12 h at 37°C, antibodies specific for p-MK2, p-MK5, MK2, MK5 were used to assess the protein levels in the cell lysates. HS, heat stress; p-, phosphorylated; t-, total; MK, mitogen-activated protein kinase-activated protein kinase; SB203580, p38 inhibitor; PD98059, ERK inhibitor; SP600125, JNK inhibitor.

    Journal: Oncology Letters

    Article Title: p38 MAPK-MK2 pathway regulates the heat-stress-induced accumulation of reactive oxygen species that mediates apoptotic cell death in glial cells

    doi: 10.3892/ol.2017.7360

    Figure Lengend Snippet: Inhibition of p38 weakens heat stress-induced phosphorylation of MK5 and MK2. F98 cells underwent pretreatment with or without the indicated inhibitors for 30 min prior to exposure to heat stress or control heat treatment. After the cells received further incubation for 12 h at 37°C, antibodies specific for p-MK2, p-MK5, MK2, MK5 were used to assess the protein levels in the cell lysates. HS, heat stress; p-, phosphorylated; t-, total; MK, mitogen-activated protein kinase-activated protein kinase; SB203580, p38 inhibitor; PD98059, ERK inhibitor; SP600125, JNK inhibitor.

    Article Snippet: Cells were pretreated with CMPD-1 (10 µM; cat. no. 263847-55-8; Tocris Bioscience, Bristol, UK), MnTBAP (10 µM; cat. no. ab141496; Abcam, Cambridge, MA, USA), SP600125 (10 µM; cat. no. ab120065; Abcam), SB203580 (5 µM; cat. no. ab120162; Abcam), PD98059 (10 µM; cat. no. ab120234; Abcam), SB202474 (10 µM; cat. no. IMA1013; Jingke Science and Technology Co., Ltd., Shanghai, China), the specific inhibitors of MK2, ROS, JNK, p38, ERK and the negative control, respectively, at 37°C for 30 min.

    Techniques: Inhibition, Incubation

    U0126 and PD98059 inhibit IL-12 production by macrophages infected with N. caninum. (A) PMϕ were infected with N. caninum -1 tachyzoites at a MOI of 10 in the presence or absence (Ctr) of UO126 and PD98059 (0–20ìM, 0ìM was treated with same amount of DMSO), respectively, in DMEM with 1% FBS at 37°C for 24 h. (B) PMϕ were incubated with U0126 and PD98059 with 20 μM for 1 h, respectively. Ctr was incubated with same amount of DMSO. Then after the inhibitor or DMSO was washed off (pre-treatment) or not (treatment), PMϕ were infected with tachyzoites at a MOI of 10 for 24 h at 37°C. Samples were applied in triplicates for ELISA assay and data are expressed as the mean of three independent experiments ± SEM. ∗ P

    Journal: Frontiers in Microbiology

    Article Title: Activation of ERK Signaling via TLR11 Induces IL-12p40 Production in Peritoneal Macrophages Challenged by Neospora caninum

    doi: 10.3389/fmicb.2017.01393

    Figure Lengend Snippet: U0126 and PD98059 inhibit IL-12 production by macrophages infected with N. caninum. (A) PMϕ were infected with N. caninum -1 tachyzoites at a MOI of 10 in the presence or absence (Ctr) of UO126 and PD98059 (0–20ìM, 0ìM was treated with same amount of DMSO), respectively, in DMEM with 1% FBS at 37°C for 24 h. (B) PMϕ were incubated with U0126 and PD98059 with 20 μM for 1 h, respectively. Ctr was incubated with same amount of DMSO. Then after the inhibitor or DMSO was washed off (pre-treatment) or not (treatment), PMϕ were infected with tachyzoites at a MOI of 10 for 24 h at 37°C. Samples were applied in triplicates for ELISA assay and data are expressed as the mean of three independent experiments ± SEM. ∗ P

    Article Snippet: For treatment with UO126 or PD98059 (Selleck Chemicals, United States), which are specific kinase inhibitors against ERK and ERK signaling upstream MEK, respectively, PMϕ were treated with tachyzoites (MOI = 10) or LPS (50 ng/ml) in the presence or absent of UO126/PD98059 at different concentrations (5–20 μM) under conditions discussed in the results section; for pre-treatment with UO126/PD98059, PMϕ were incubated with the inhibitor (20 μM) or DMSO for 1 h and after washing with sterile PBS, tachyzoites or LPS were added; for treatment with monoclonal antibodies against TLR11 (ab47097, Abcam, United States) , PMϕ were incubated with the antibodies at the concentration of 3 μg/ml or same amount of homologous negative serum for 2 h at 37°C before tachyzoites (MOI = 10) were added to the cells for 1, 24, and 48 h, respectively; for treatment with siRNAs, PMϕ were inoculated with tachyzoites (MOI = 10) for 1 and 24 h, respectively, after transfection with Myd88 siRNA or control siRNA for 36 h. After different treatments, supernatants and cells were collected for ELISA and Western blotting.

    Techniques: Infection, Incubation, Enzyme-linked Immunosorbent Assay

    Nrf2 signaling pathway regulates MAPK cascade. ( a ) THP-1-derived macrophages were pretreated with MAPK-specific inhibitors PD98059, SB203580, and SP600125 of ERK, p38, and JNK, respectively, 2 h before SFN stimulation and/or Mabs infection. Cells were lysed 24 h after infection and protein lysates were analyzed by western blot. ( b ) THP-1-derived macrophages were transfected with scrambled or Nrf2 siRNA 24 h before SFN stimulation and/or M. abscessus infection. Cells were incubated for an additional 24 h after infection before lysis and western blots were performed using Nrf2 antibody and GAPDH antibody, as an internal control. ( c ) Immunoblots were performed on total protein lysates using specific phosphorylated antibodies against ERK, JNK, and p38.

    Journal: Cell Death Discovery

    Article Title: Caspase-independent apoptosis in infected macrophages triggered by sulforaphane via Nrf2/p38 signaling pathways

    doi: 10.1038/cddiscovery.2015.22

    Figure Lengend Snippet: Nrf2 signaling pathway regulates MAPK cascade. ( a ) THP-1-derived macrophages were pretreated with MAPK-specific inhibitors PD98059, SB203580, and SP600125 of ERK, p38, and JNK, respectively, 2 h before SFN stimulation and/or Mabs infection. Cells were lysed 24 h after infection and protein lysates were analyzed by western blot. ( b ) THP-1-derived macrophages were transfected with scrambled or Nrf2 siRNA 24 h before SFN stimulation and/or M. abscessus infection. Cells were incubated for an additional 24 h after infection before lysis and western blots were performed using Nrf2 antibody and GAPDH antibody, as an internal control. ( c ) Immunoblots were performed on total protein lysates using specific phosphorylated antibodies against ERK, JNK, and p38.

    Article Snippet: MAPK inhibitors PD98059, SB203580, SP600125, annexin-V FITC apoptosis detection kit, anti-lamin A/C, anti-β actin, and anti-HO-1 antibodies were purchased from Abcam (Cambridge, UK).

    Techniques: Derivative Assay, Infection, Western Blot, Transfection, Incubation, Lysis

    ( a ) Annexin V-FITC assay in THP-1-derived macrophages pretreated with MAPK inhibitors SB203580, SP600125, and PD98059. Infected cells were collected 24 h post infection. Data were from two independent experiments of n =10 000 events that were imaged and analyzed using MARKII imaging cytometer. * P

    Journal: Cell Death Discovery

    Article Title: Caspase-independent apoptosis in infected macrophages triggered by sulforaphane via Nrf2/p38 signaling pathways

    doi: 10.1038/cddiscovery.2015.22

    Figure Lengend Snippet: ( a ) Annexin V-FITC assay in THP-1-derived macrophages pretreated with MAPK inhibitors SB203580, SP600125, and PD98059. Infected cells were collected 24 h post infection. Data were from two independent experiments of n =10 000 events that were imaged and analyzed using MARKII imaging cytometer. * P

    Article Snippet: MAPK inhibitors PD98059, SB203580, SP600125, annexin-V FITC apoptosis detection kit, anti-lamin A/C, anti-β actin, and anti-HO-1 antibodies were purchased from Abcam (Cambridge, UK).

    Techniques: Derivative Assay, Infection, Imaging, Cytometry

    ERK1/2 inhibitor PD98059 prevents the increases in surface levels of both GluA1 and GluA2 subunits induced by LY37 treatment in cultured PFC neurons. (A) Representative images of drug treatment. Primary cultured PFC neurons at 17–18 DIV were exposed to DMSO (0.02%), LY37 (1 µM), ERK inhibitor PD98059 (50 µM), or PD98059 (50 µM) + LY37 (1 µM) for 1 h at 37°C, respectively. After treatments, the cells were incubated in BS 3 cross-linker and the protein levels on the membrane surface and intracellular components were then separated by Western blot. (B and C) Summary bar graphs show the protein levels of surface and intracellular GluA1 and GluA2 subunits in different treatments groups compared to vehicle controls. Neither ERK1/2 inhibitor PD98059 nor PD98059+LY37 treatment induced changes in GluA1 and GluA2 surface expression (p > 0.05 for all comparisons).

    Journal: PLoS ONE

    Article Title: Group II Metabotropic Glutamate Receptor Agonist LY379268 Regulates AMPA Receptor Trafficking in Prefrontal Cortical Neurons

    doi: 10.1371/journal.pone.0061787

    Figure Lengend Snippet: ERK1/2 inhibitor PD98059 prevents the increases in surface levels of both GluA1 and GluA2 subunits induced by LY37 treatment in cultured PFC neurons. (A) Representative images of drug treatment. Primary cultured PFC neurons at 17–18 DIV were exposed to DMSO (0.02%), LY37 (1 µM), ERK inhibitor PD98059 (50 µM), or PD98059 (50 µM) + LY37 (1 µM) for 1 h at 37°C, respectively. After treatments, the cells were incubated in BS 3 cross-linker and the protein levels on the membrane surface and intracellular components were then separated by Western blot. (B and C) Summary bar graphs show the protein levels of surface and intracellular GluA1 and GluA2 subunits in different treatments groups compared to vehicle controls. Neither ERK1/2 inhibitor PD98059 nor PD98059+LY37 treatment induced changes in GluA1 and GluA2 surface expression (p > 0.05 for all comparisons).

    Article Snippet: To detect surface and intracellular protein, culture neurons at DIV17–18 were treated with LY37 (1 µM), or ERK1/2 inhibitor PD98059 (50 µM, Abcam, Cambridge, MA) for 1 h prior to treatment with LY37 (1 µM).

    Techniques: Cell Culture, Incubation, Western Blot, Expressing

    LY37 treatment significantly increased the phosphorylation of ERK1/2 activity in the cultured PFC neurons, and this was prevented by the ERK inhibitor PD98059 (PD). (A) Representative images of ERK1/2 and p-ERK1/2 expression in response to LY37, PD (50 µM for 1 h) or PD followed by LY37 treatment (1 µM for 1 h), DMSO as PD's vehicle. (B) LY37 administration did not induce clear changes in either ERK1 or ERK2 total protein level (p > 0.05). However, it significantly increased the expression of both p-ERK1 and p-ERK2 ratios (**p

    Journal: PLoS ONE

    Article Title: Group II Metabotropic Glutamate Receptor Agonist LY379268 Regulates AMPA Receptor Trafficking in Prefrontal Cortical Neurons

    doi: 10.1371/journal.pone.0061787

    Figure Lengend Snippet: LY37 treatment significantly increased the phosphorylation of ERK1/2 activity in the cultured PFC neurons, and this was prevented by the ERK inhibitor PD98059 (PD). (A) Representative images of ERK1/2 and p-ERK1/2 expression in response to LY37, PD (50 µM for 1 h) or PD followed by LY37 treatment (1 µM for 1 h), DMSO as PD's vehicle. (B) LY37 administration did not induce clear changes in either ERK1 or ERK2 total protein level (p > 0.05). However, it significantly increased the expression of both p-ERK1 and p-ERK2 ratios (**p

    Article Snippet: To detect surface and intracellular protein, culture neurons at DIV17–18 were treated with LY37 (1 µM), or ERK1/2 inhibitor PD98059 (50 µM, Abcam, Cambridge, MA) for 1 h prior to treatment with LY37 (1 µM).

    Techniques: Activity Assay, Cell Culture, Expressing

    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

    sCLU regulates the cisplatin (DDP)-induced cytotoxicity in the osteosarcoma (OS) cells. (A) The KH OS or sCLU stably transfected KH OS cells (KH OS/sCLU) were treated with 0.01 to 10 μM/ mL DPP for 72 hours in the presence or absence of 10 mM PD98059. Cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results were obtained from three independent experiments. Versus KHOS/sCLU, * P

    Journal: World Journal of Surgical Oncology

    Article Title: Secreted clusterin (sCLU) regulates cell proliferation and chemosensitivity to cisplatin by modulating ERK1/2 signals in human osteosarcoma cells

    doi: 10.1186/1477-7819-12-255

    Figure Lengend Snippet: sCLU regulates the cisplatin (DDP)-induced cytotoxicity in the osteosarcoma (OS) cells. (A) The KH OS or sCLU stably transfected KH OS cells (KH OS/sCLU) were treated with 0.01 to 10 μM/ mL DPP for 72 hours in the presence or absence of 10 mM PD98059. Cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The results were obtained from three independent experiments. Versus KHOS/sCLU, * P

    Article Snippet: PD98059, a specific inhibitor of ERK kinase was purchased from Biovision, Beijing, China.

    Techniques: Stable Transfection, Transfection, MTT Assay