mek inhibitor Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore mitogen activated protein kinase extracellular signal regulated kinase kinase mek inhibitor pd98059
    Effect of <t>MEK-ERK</t> inhibition on the density profile of secreted apoB100 containing lipoproteins. HepG2 cells and Huh-7 cells were pretreated with 5 μmol/l <t>PD98059</t> and metabolically labeled to steady-state with [ 35 S]methionine/cysteine in the presence
    Mitogen Activated Protein Kinase Extracellular Signal Regulated Kinase Kinase Mek Inhibitor Pd98059, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein kinase extracellular signal regulated kinase kinase mek inhibitor pd98059/product/Millipore
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein kinase extracellular signal regulated kinase kinase mek inhibitor pd98059 - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    86
    Millipore mitogen activated protein extracellular signal regulated kinase mek inhibitors u0126
    Effect of <t>MEK-ERK</t> inhibition on the density profile of secreted apoB100 containing lipoproteins. HepG2 cells and Huh-7 cells were pretreated with 5 μmol/l <t>PD98059</t> and metabolically labeled to steady-state with [ 35 S]methionine/cysteine in the presence
    Mitogen Activated Protein Extracellular Signal Regulated Kinase Mek Inhibitors U0126, supplied by Millipore, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein extracellular signal regulated kinase mek inhibitors u0126/product/Millipore
    Average 86 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein extracellular signal regulated kinase mek inhibitors u0126 - by Bioz Stars, 2020-05
    86/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc mitogen activated protein extracellular signal regulated kinase kinase mek 1 2 inhibitor
    Effect of <t>MEK-ERK</t> inhibition on the density profile of secreted apoB100 containing lipoproteins. HepG2 cells and Huh-7 cells were pretreated with 5 μmol/l <t>PD98059</t> and metabolically labeled to steady-state with [ 35 S]methionine/cysteine in the presence
    Mitogen Activated Protein Extracellular Signal Regulated Kinase Kinase Mek 1 2 Inhibitor, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein extracellular signal regulated kinase kinase mek 1 2 inhibitor/product/Cell Signaling Technology Inc
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein extracellular signal regulated kinase kinase mek 1 2 inhibitor - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    99
    Tocris mitogen activated protein kinase extracellular signal regulated kinase mek erk inhibitor
    Potential signaling pathways implicated in the spontaneous motility of neurospheres. (A~D) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were taken at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Neurospheres were treat with control (CTL) PBS (A), a PI3K inhibitor LY294002 (10 µM) (B), an <t>MEK/ERK</t> inhibitor U0126 (10 µM) (C), and a ROCK inhibitor Y27632 (10 µg/ml) (D). Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Scale bars indicate 100 µm. (E, F) Quantification graphs of the moving distance (E) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (F) growing from the tracked neurospheres. Total cumulative distance or length traced for the entire culture duration was plotted. N=15 neurospheres tracked from three independent cultures. * and *** represent p
    Mitogen Activated Protein Kinase Extracellular Signal Regulated Kinase Mek Erk Inhibitor, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein kinase extracellular signal regulated kinase mek erk inhibitor/product/Tocris
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein kinase extracellular signal regulated kinase mek erk inhibitor - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    90
    Millipore mitogen activated protein kinase extracellular signal regulated kinase inhibitor
    Potential signaling pathways implicated in the spontaneous motility of neurospheres. (A~D) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were taken at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Neurospheres were treat with control (CTL) PBS (A), a PI3K inhibitor LY294002 (10 µM) (B), an <t>MEK/ERK</t> inhibitor U0126 (10 µM) (C), and a ROCK inhibitor Y27632 (10 µg/ml) (D). Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Scale bars indicate 100 µm. (E, F) Quantification graphs of the moving distance (E) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (F) growing from the tracked neurospheres. Total cumulative distance or length traced for the entire culture duration was plotted. N=15 neurospheres tracked from three independent cultures. * and *** represent p
    Mitogen Activated Protein Kinase Extracellular Signal Regulated Kinase Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein kinase extracellular signal regulated kinase inhibitor/product/Millipore
    Average 90 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein kinase extracellular signal regulated kinase inhibitor - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    85
    Cayman Chemical mitogen activated protein extracellular signal regulated kinase mek inhibitor pd0325901
    Potential signaling pathways implicated in the spontaneous motility of neurospheres. (A~D) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were taken at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Neurospheres were treat with control (CTL) PBS (A), a PI3K inhibitor LY294002 (10 µM) (B), an <t>MEK/ERK</t> inhibitor U0126 (10 µM) (C), and a ROCK inhibitor Y27632 (10 µg/ml) (D). Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Scale bars indicate 100 µm. (E, F) Quantification graphs of the moving distance (E) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (F) growing from the tracked neurospheres. Total cumulative distance or length traced for the entire culture duration was plotted. N=15 neurospheres tracked from three independent cultures. * and *** represent p
    Mitogen Activated Protein Extracellular Signal Regulated Kinase Mek Inhibitor Pd0325901, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein extracellular signal regulated kinase mek inhibitor pd0325901/product/Cayman Chemical
    Average 85 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein extracellular signal regulated kinase mek inhibitor pd0325901 - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    85
    Promega mitogen activated protein extracellular signal regulated kinase mek inhibitor u0126
    Nuclear factor-κB (NF-κB) inhibitor dimethylfumarate (DMF) abrogated TNF-α- and IL-1β-induced nuclear translocation of NF-κB p65 subunit and IL-33 expression and mitogen-activated protein/extracellular signal-regulated kinase <t>(MEK)</t> or janus-activated kinase (JAK) 1 and 2 inhibitors reduced TNF-α, IFN-γ, or IL-1β-induced IL-33 expression in human adult cardiac fibroblasts (HACF), cardiac myocytes (HACM), or coronary artery smooth muscle cells (HCASMC). HACF, HACM, and HCASMC were pre-incubated for 30 min with DMF at 100 μM. Subsequently, these cells were treated with TNF-α- or IL-1β (each at 2000 U/mL) for 60 min. Preparation of nuclear extracts and quantification of p65 NF-κB subunit (A) were performed as described in “ Materials and methods ”. Values are given as OD492 nm and represent mean ± SD. Experiments were performed 2 times with cells obtained from 2 different donors for each cell types. HACF, HACM, and HCASMC were first pre-incubated with DMF at 100 μM for 30 min and then treated with TNF-α- or IL-1β for 24 h (B). HACF, HACM, and HCASMC were first pre-incubated with <t>U0126</t> at 10 μM for 30 min and then treated with TNF-α, IFN-γ or IL-1β for 24 h (C). HACM were first pre-incubated with JAK inhibitor I at 10 μM for 30 min and then treated with IFN-γ at 2000 U/mL for 24 h (D). IL-33 protein in the cell lysates was measured by a specific ELISA as described in “ Materials and methods ”. Each experiment was performed in triplicates. Values are given in IL-33 (units) as % of control, which was set as 100% and represent mean ± SD. Experiments were performed 3 times with cells obtained from 3 different donors. *p ≤ 0.05 as compared to untreated control in HACF; **p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACF; §p ≤ 0.05 as compare to untreated control in HACM; §§p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACM; $p ≤ 0.05 as compared to untreated control in HCASMC; $$p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HCASMC.
    Mitogen Activated Protein Extracellular Signal Regulated Kinase Mek Inhibitor U0126, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein extracellular signal regulated kinase mek inhibitor u0126/product/Promega
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein extracellular signal regulated kinase mek inhibitor u0126 - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    98
    Millipore mitogen activated protein extracellular signal regulated kinase 1 2 mek1 2 inhibitors u0126
    Nuclear factor-κB (NF-κB) inhibitor dimethylfumarate (DMF) abrogated TNF-α- and IL-1β-induced nuclear translocation of NF-κB p65 subunit and IL-33 expression and mitogen-activated protein/extracellular signal-regulated kinase <t>(MEK)</t> or janus-activated kinase (JAK) 1 and 2 inhibitors reduced TNF-α, IFN-γ, or IL-1β-induced IL-33 expression in human adult cardiac fibroblasts (HACF), cardiac myocytes (HACM), or coronary artery smooth muscle cells (HCASMC). HACF, HACM, and HCASMC were pre-incubated for 30 min with DMF at 100 μM. Subsequently, these cells were treated with TNF-α- or IL-1β (each at 2000 U/mL) for 60 min. Preparation of nuclear extracts and quantification of p65 NF-κB subunit (A) were performed as described in “ Materials and methods ”. Values are given as OD492 nm and represent mean ± SD. Experiments were performed 2 times with cells obtained from 2 different donors for each cell types. HACF, HACM, and HCASMC were first pre-incubated with DMF at 100 μM for 30 min and then treated with TNF-α- or IL-1β for 24 h (B). HACF, HACM, and HCASMC were first pre-incubated with <t>U0126</t> at 10 μM for 30 min and then treated with TNF-α, IFN-γ or IL-1β for 24 h (C). HACM were first pre-incubated with JAK inhibitor I at 10 μM for 30 min and then treated with IFN-γ at 2000 U/mL for 24 h (D). IL-33 protein in the cell lysates was measured by a specific ELISA as described in “ Materials and methods ”. Each experiment was performed in triplicates. Values are given in IL-33 (units) as % of control, which was set as 100% and represent mean ± SD. Experiments were performed 3 times with cells obtained from 3 different donors. *p ≤ 0.05 as compared to untreated control in HACF; **p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACF; §p ≤ 0.05 as compare to untreated control in HACM; §§p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACM; $p ≤ 0.05 as compared to untreated control in HCASMC; $$p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HCASMC.
    Mitogen Activated Protein Extracellular Signal Regulated Kinase 1 2 Mek1 2 Inhibitors U0126, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitogen activated protein extracellular signal regulated kinase 1 2 mek1 2 inhibitors u0126/product/Millipore
    Average 98 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    mitogen activated protein extracellular signal regulated kinase 1 2 mek1 2 inhibitors u0126 - by Bioz Stars, 2020-05
    98/100 stars
      Buy from Supplier

    92
    Biomol GmbH mek inhibitor
    MIP-1β interfere with monocyte differentiation into DC. Monocytes isolated from an healthy individual were differentiated into DC in presence or not of 200 ng/ml MIP-1β. When indicated, monocytes were treated with the <t>MEK</t> inhibitor (PD) at day 4 and maturation was induced by LPS at day 5. Cells harvested at day 6, were cocultured for 5 days with allogeneic T cells at a ratio of 1/10 DC/T cell in triplicates. <t>IFNγ</t> was measured by CBA in supernatant of cocultures. Mean IFNγ secretion±SD of triplicate wells is shown. * The mean IFNγ secretion stimulated by Mo-DC treated with MIP-1β and LPS is significantly reduced compared to that of MoDC treated with LPS with p
    Mek Inhibitor, supplied by Biomol GmbH, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Biomol GmbH
    Average 92 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Cell Signaling Technology Inc mek inhibitor
    Effects of <t>MEK</t> or PI3K inhibitors on the production of MMP-3, VEGF and PGE2 by MH7A cells. A . MH7A cells (2 x 10 4 cells/well) were cultured with the MEK inhibitor, <t>PD98059</t> (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. B . MH7A cells (2 x 10 4 cells/well) were cultured with the PI3K inhibitor, LY294002 (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. Data represent the mean ± SD of three independent experiments. *P
    Mek Inhibitor, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 92/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Cell Signaling Technology Inc
    Average 92 stars, based on 127 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    FUJIFILM mek inhibitor
    Detection of pERK signaling of vascular endothelial cells in Vegfr3-Gap43-Venus Tg embryos at <t>E9.5.</t> (A) Flow-cytometry analysis of Vegfr3-Gap43-Venus Tg embryos at E9.5. (B) Western blot analysis of Venus-positive and -negative cells from Tg embryos. (C) Phosphorylated ERK signaling in Venus-positive cells cultured in the presence or absence of the <t>MEK</t> inhibitor. (D) Percentage of pERK-positive cells in Venus-positive cells.
    Mek Inhibitor, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/FUJIFILM
    Average 92 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Millipore mek inhibitor
    DAZAP1 regulates cell proliferation (a) Stable over-expression of DAZAP1 in non-small cell lung carcinoma NCI-H157 cells as detected by western blots. Control cells are stably transfected with vector only. (b) The expression of DAZAP1 inhibited cell proliferation as judged by colony formation assay with H157 cells. A representative picture from the triplicate experiments, the means and s.d. were plotted. (c) DAZAP1 inhibited the anchorage independent cell growth of H157 as judged by soft agar assays. A representative picture from the triplicates and the quantification were shown. The box plot was drawn using 25 and 75 quartiles in <t>Sigma</t> Plot with whiskers indicating max and min value and dots indicating outliers. P values were determined using paired t-test, *=p ≤ 5.0×10 −5 . (d) DAZAP1 inhibited cell migration as judged by wound healing assay. The picture of a representative plate (left panel) and the quantification of percent of gap closure were shown (right panel). Data from three independent assays were performed error bar indicate s.d. (e) The quantification of cell cycle stages (G1, S and G2/M phases) in DAZAP1 over-expressed or control cells. The cell cycle stages were analyzed by flow-cytometry measurement of DNA content. Data from two independent flow analysis, error bar indicate s.d. (f) Expression of endogenous DAZAP1 protein during cell cycle as detected by western blots. Tubulin and canonical cell cycle markers (cyclin A1 and B1) were also probed as controls. (g) An integrated model for DAZAP1 function and the regulation of its activity through the <t>MEK/Erk</t> pathway.
    Mek Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 324 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Millipore
    Average 92 stars, based on 324 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Pfizer Inc mek inhibitor
    Rationale for Targeting Both the <t>Ras/Raf/MEK/ERK</t> and Ras/PI3K/PTEN/Akt/mTOR Pathways for Suppressing Cancer Growth A: The Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways are both activated by upstream receptor ligation and frequently co-regulate many downstream targets in parallel. Thus for effective elimination of many cancers or prevention of aging, it may be necessary to target both signaling pathways. Activation of these pathways could also result in increased transcription of many genes that promote cellular growth and malignant transformation. B. Inhibition of mTOR can result in the induction of autophagy, which is a very important mechanism of cell death, especially in solid tumors. C. As described previously, both the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways regulate the activity of apoptotic proteins by post-translational mechanisms. Targeting this pathway may also contribute to the induction of <t>apoptosis.</t> Signaling molecules promoting phosphorylation events are indicated in green. Stimulatory signaling events are indicted in green lines with a green arrow before the target of the phophorylation. Small molecule inhibitors are indicated in red. Inhibitory phosphorylation events are indicated in red lines with a block on the end before the target of the inhibition. Inhibitory signaling or proapoptotic molecules or inactivated molecules are indicated in yellow. A growth factor and a growth factor receptor are indicated in purple. Active transcription factors are indicated in purple diamonds. Inactivated transcription factors are indicated in yellow diamonds.
    Mek Inhibitor, supplied by Pfizer Inc, used in various techniques. Bioz Stars score: 92/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Pfizer Inc
    Average 92 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Stemgent mek inhibitor
    Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 <t>(MEK</t> inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories
    Mek Inhibitor, supplied by Stemgent, used in various techniques. Bioz Stars score: 92/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Stemgent
    Average 92 stars, based on 59 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    DuPont de Nemours mek inhibitor
    Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 <t>(MEK</t> inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories
    Mek Inhibitor, supplied by DuPont de Nemours, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/DuPont de Nemours
    Average 92 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    92
    Promega mek inhibitor
    <t>MEK</t> inhibitor inhibits CVB3-mediated RasGAP cleavage. HeLa cells were pretreated for 30 min with various concentrations of <t>U0126</t> and then were infected by CVB3 for 1 h. At 9 h after addition of CVB3, HeLa cells were harvested and Western blot analysis was performed using an antibody that recognizes RasGAP. Results were quantitated by densitometric analysis using National Institutes of Health Image, version 1.61, and normalized to the control level (sham-infected cells without U0126), which was arbitrarily set to 1.0. Values are means ± standard errors ( n = 3).
    Mek Inhibitor, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 146 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Promega
    Average 92 stars, based on 146 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    91
    Cayman Chemical mek inhibitor
    The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and <t>U0126</t> <t>MEK</t> inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P
    Mek Inhibitor, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 91/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Cayman Chemical
    Average 91 stars, based on 36 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    92
    Fisher Scientific mek inhibitor
    The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and <t>U0126</t> <t>MEK</t> inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P
    Mek Inhibitor, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Fisher Scientific
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    91
    Genentech mek inhibitor
    The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and <t>U0126</t> <t>MEK</t> inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P
    Mek Inhibitor, supplied by Genentech, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Genentech
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    91
    InvivoGen mek inhibitor
    The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and <t>U0126</t> <t>MEK</t> inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P
    Mek Inhibitor, supplied by InvivoGen, used in various techniques. Bioz Stars score: 91/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/InvivoGen
    Average 91 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    93
    Selleck Chemicals mek inhibitor
    <t>MEK</t> inhibitor suppressed spheroid formation in <t>ID8-KRAS</t> cells. ID8-KRAS-GFP cells (2 × 10 5 ) were not treated (0 nM) or treated with trametinib (1, 10, 100 nM) for 48 h in 2D or 3D culture. a Representative microscopic image of cancer cells under 2D and 3D conditions at 100× magnification. b-c After the treatment, cells were exposed to EdU (10 μM) for 2 h and fixed. EdU was labeled with Alexa Fluor 647. EdU uptake was analyzed ( b ) and the ratio of EdU-positive to EdU-negative cells was measured by flow cytometry ( c ). Experiments were repeated at least thrice. The values shown represent mean ± SEM (* p
    Mek Inhibitor, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 93/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Selleck Chemicals
    Average 93 stars, based on 170 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    92
    Pharmingen mek inhibitor
    <t>MEK</t> inhibitor suppressed spheroid formation in <t>ID8-KRAS</t> cells. ID8-KRAS-GFP cells (2 × 10 5 ) were not treated (0 nM) or treated with trametinib (1, 10, 100 nM) for 48 h in 2D or 3D culture. a Representative microscopic image of cancer cells under 2D and 3D conditions at 100× magnification. b-c After the treatment, cells were exposed to EdU (10 μM) for 2 h and fixed. EdU was labeled with Alexa Fluor 647. EdU uptake was analyzed ( b ) and the ratio of EdU-positive to EdU-negative cells was measured by flow cytometry ( c ). Experiments were repeated at least thrice. The values shown represent mean ± SEM (* p
    Mek Inhibitor, supplied by Pharmingen, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor/product/Pharmingen
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    90
    Stemgent mek inhibitor pd0325901
    Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM <t>PD0325901</t> <t>(MEK</t> inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories
    Mek Inhibitor Pd0325901, supplied by Stemgent, used in various techniques. Bioz Stars score: 90/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mek inhibitor pd0325901/product/Stemgent
    Average 90 stars, based on 98 article reviews
    Price from $9.99 to $1999.99
    mek inhibitor pd0325901 - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    Image Search Results


    Effect of MEK-ERK inhibition on the density profile of secreted apoB100 containing lipoproteins. HepG2 cells and Huh-7 cells were pretreated with 5 μmol/l PD98059 and metabolically labeled to steady-state with [ 35 S]methionine/cysteine in the presence

    Journal: Journal of Lipid Research

    Article Title: Huh-7 or HepG2 cells: which is the better model for studying human apolipoprotein-B100 assembly and secretion? [S]

    doi: 10.1194/jlr.D008888

    Figure Lengend Snippet: Effect of MEK-ERK inhibition on the density profile of secreted apoB100 containing lipoproteins. HepG2 cells and Huh-7 cells were pretreated with 5 μmol/l PD98059 and metabolically labeled to steady-state with [ 35 S]methionine/cysteine in the presence

    Article Snippet: The mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK-ERK) inhibitor PD98059 was purchased from Calbiochem (San Diego, CA).

    Techniques: Inhibition, Metabolic Labelling, Labeling

    IGF-1 activates the PI3K/Akt and MEK/ERK signaling pathways. A, IGF-1 increases the phosphorylation of Akt at 30 min and (B) ERK at 10 min. IGF-1 treatment preferentially induced phosphorylation of ERK2 (p42), while phospho-ERK1 (p44) signal was barely

    Journal: The Journal of Neuroscience

    Article Title: IGF-1 promotes G1/S cell cycle progression through bidirectional regulation of cyclins and CDK inhibitors via the PI3K/Akt pathway in developing rat cerebral cortex

    doi: 10.1523/JNEUROSCI.1700-08.2009

    Figure Lengend Snippet: IGF-1 activates the PI3K/Akt and MEK/ERK signaling pathways. A, IGF-1 increases the phosphorylation of Akt at 30 min and (B) ERK at 10 min. IGF-1 treatment preferentially induced phosphorylation of ERK2 (p42), while phospho-ERK1 (p44) signal was barely

    Article Snippet: Transferrin, PI3K inhibitor (LY294002; 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) and MEK inhibitors (PD98059, 2-Amino-3-methoxyflavone; U0126, 1,4-Diamino-2,3-dicyano-1,4- bis (2-aminophenylthio)butadiene) were obtained from Calbiochem (La Jolla, CA).

    Techniques:

    Effects of IGF-1 ICV injections on the PI3K/Akt and MEK/ERK pathways. A, IGF-1 injections into the lateral ventricles of E16.5 rat embryos (30 ng per embryo) resulted in a 4 fold increase in the phosphorylation of Akt by 30 min, revealed by Western blotting.

    Journal: The Journal of Neuroscience

    Article Title: IGF-1 promotes G1/S cell cycle progression through bidirectional regulation of cyclins and CDK inhibitors via the PI3K/Akt pathway in developing rat cerebral cortex

    doi: 10.1523/JNEUROSCI.1700-08.2009

    Figure Lengend Snippet: Effects of IGF-1 ICV injections on the PI3K/Akt and MEK/ERK pathways. A, IGF-1 injections into the lateral ventricles of E16.5 rat embryos (30 ng per embryo) resulted in a 4 fold increase in the phosphorylation of Akt by 30 min, revealed by Western blotting.

    Article Snippet: Transferrin, PI3K inhibitor (LY294002; 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) and MEK inhibitors (PD98059, 2-Amino-3-methoxyflavone; U0126, 1,4-Diamino-2,3-dicyano-1,4- bis (2-aminophenylthio)butadiene) were obtained from Calbiochem (La Jolla, CA).

    Techniques: Western Blot

    Blockade of PI3K/Akt signaling, but not the MEK/ERK pathway, inhibits IGF-1 stimulatory effects on DNA synthesis of cortical precursors. A, Cells were pre-incubated with PI3K inhibitor LY294002 (LY) for 30 min before IGF-1 addition, and DNA synthesis

    Journal: The Journal of Neuroscience

    Article Title: IGF-1 promotes G1/S cell cycle progression through bidirectional regulation of cyclins and CDK inhibitors via the PI3K/Akt pathway in developing rat cerebral cortex

    doi: 10.1523/JNEUROSCI.1700-08.2009

    Figure Lengend Snippet: Blockade of PI3K/Akt signaling, but not the MEK/ERK pathway, inhibits IGF-1 stimulatory effects on DNA synthesis of cortical precursors. A, Cells were pre-incubated with PI3K inhibitor LY294002 (LY) for 30 min before IGF-1 addition, and DNA synthesis

    Article Snippet: Transferrin, PI3K inhibitor (LY294002; 2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) and MEK inhibitors (PD98059, 2-Amino-3-methoxyflavone; U0126, 1,4-Diamino-2,3-dicyano-1,4- bis (2-aminophenylthio)butadiene) were obtained from Calbiochem (La Jolla, CA).

    Techniques: DNA Synthesis, Incubation

    Potential signaling pathways implicated in the spontaneous motility of neurospheres. (A~D) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were taken at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Neurospheres were treat with control (CTL) PBS (A), a PI3K inhibitor LY294002 (10 µM) (B), an MEK/ERK inhibitor U0126 (10 µM) (C), and a ROCK inhibitor Y27632 (10 µg/ml) (D). Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Scale bars indicate 100 µm. (E, F) Quantification graphs of the moving distance (E) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (F) growing from the tracked neurospheres. Total cumulative distance or length traced for the entire culture duration was plotted. N=15 neurospheres tracked from three independent cultures. * and *** represent p

    Journal: Experimental Neurobiology

    Article Title: Insulin-like Growth Factor-1 Receptor Dictates Beneficial Effects of Treadmill Training by Regulating Survival and Migration of Neural Stem Cell Grafts in the Injured Spinal Cord

    doi: 10.5607/en.2018.27.6.489

    Figure Lengend Snippet: Potential signaling pathways implicated in the spontaneous motility of neurospheres. (A~D) Representative still images of cultured neurospheres obtained using a live cell imaging apparatus. Images were taken at the time points designated at the upper right corners. 0 h indicates a start of image tracking for the next 12 h, not the absolute start of the culture. Neurospheres were treat with control (CTL) PBS (A), a PI3K inhibitor LY294002 (10 µM) (B), an MEK/ERK inhibitor U0126 (10 µM) (C), and a ROCK inhibitor Y27632 (10 µg/ml) (D). Grid lines were drawn to facilitate tracking positions of neurospheres. Colored dots indicate the marks of neurosphere centers that are traced to quantify distances of neurosphere movements. Neurospheres marked with dots of the same color in the images at different time points are the identical ones. Scale bars indicate 100 µm. (E, F) Quantification graphs of the moving distance (E) of the tracked neurospheres and the length of changes in cytoplasmic protrusions (F) growing from the tracked neurospheres. Total cumulative distance or length traced for the entire culture duration was plotted. N=15 neurospheres tracked from three independent cultures. * and *** represent p

    Article Snippet: To examine the influence of IGF-1 and related signaling pathways on the neurosphere motility, IGF-1 (50 ng/ml, #791-MG, R & D Systems), phosphoinositide-3 kinase (PI3K) inhibitor (LY294002; 10 µM, #440202, CalbioChem), mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK/ERK) inhibitor (U0126; 10 µM, #662005, Calbio-Chem), Rho-associated protein kinase (ROCK) inhibitor (Y27632; 10 µg/ml, #129830-38-2, Tocris), IGF-1R inhibitor (PPP, picropodophyllin; 100 µM, #SC-204008, Santa Cruz Biotechnology) were added in culture medium.

    Techniques: Cell Culture, Live Cell Imaging, CTL Assay

    Nuclear factor-κB (NF-κB) inhibitor dimethylfumarate (DMF) abrogated TNF-α- and IL-1β-induced nuclear translocation of NF-κB p65 subunit and IL-33 expression and mitogen-activated protein/extracellular signal-regulated kinase (MEK) or janus-activated kinase (JAK) 1 and 2 inhibitors reduced TNF-α, IFN-γ, or IL-1β-induced IL-33 expression in human adult cardiac fibroblasts (HACF), cardiac myocytes (HACM), or coronary artery smooth muscle cells (HCASMC). HACF, HACM, and HCASMC were pre-incubated for 30 min with DMF at 100 μM. Subsequently, these cells were treated with TNF-α- or IL-1β (each at 2000 U/mL) for 60 min. Preparation of nuclear extracts and quantification of p65 NF-κB subunit (A) were performed as described in “ Materials and methods ”. Values are given as OD492 nm and represent mean ± SD. Experiments were performed 2 times with cells obtained from 2 different donors for each cell types. HACF, HACM, and HCASMC were first pre-incubated with DMF at 100 μM for 30 min and then treated with TNF-α- or IL-1β for 24 h (B). HACF, HACM, and HCASMC were first pre-incubated with U0126 at 10 μM for 30 min and then treated with TNF-α, IFN-γ or IL-1β for 24 h (C). HACM were first pre-incubated with JAK inhibitor I at 10 μM for 30 min and then treated with IFN-γ at 2000 U/mL for 24 h (D). IL-33 protein in the cell lysates was measured by a specific ELISA as described in “ Materials and methods ”. Each experiment was performed in triplicates. Values are given in IL-33 (units) as % of control, which was set as 100% and represent mean ± SD. Experiments were performed 3 times with cells obtained from 3 different donors. *p ≤ 0.05 as compared to untreated control in HACF; **p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACF; §p ≤ 0.05 as compare to untreated control in HACM; §§p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACM; $p ≤ 0.05 as compared to untreated control in HCASMC; $$p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HCASMC.

    Journal: Journal of Molecular and Cellular Cardiology

    Article Title: Components of the interleukin-33/ST2 system are differentially expressed and regulated in human cardiac cells and in cells of the cardiac vasculature

    doi: 10.1016/j.yjmcc.2013.03.020

    Figure Lengend Snippet: Nuclear factor-κB (NF-κB) inhibitor dimethylfumarate (DMF) abrogated TNF-α- and IL-1β-induced nuclear translocation of NF-κB p65 subunit and IL-33 expression and mitogen-activated protein/extracellular signal-regulated kinase (MEK) or janus-activated kinase (JAK) 1 and 2 inhibitors reduced TNF-α, IFN-γ, or IL-1β-induced IL-33 expression in human adult cardiac fibroblasts (HACF), cardiac myocytes (HACM), or coronary artery smooth muscle cells (HCASMC). HACF, HACM, and HCASMC were pre-incubated for 30 min with DMF at 100 μM. Subsequently, these cells were treated with TNF-α- or IL-1β (each at 2000 U/mL) for 60 min. Preparation of nuclear extracts and quantification of p65 NF-κB subunit (A) were performed as described in “ Materials and methods ”. Values are given as OD492 nm and represent mean ± SD. Experiments were performed 2 times with cells obtained from 2 different donors for each cell types. HACF, HACM, and HCASMC were first pre-incubated with DMF at 100 μM for 30 min and then treated with TNF-α- or IL-1β for 24 h (B). HACF, HACM, and HCASMC were first pre-incubated with U0126 at 10 μM for 30 min and then treated with TNF-α, IFN-γ or IL-1β for 24 h (C). HACM were first pre-incubated with JAK inhibitor I at 10 μM for 30 min and then treated with IFN-γ at 2000 U/mL for 24 h (D). IL-33 protein in the cell lysates was measured by a specific ELISA as described in “ Materials and methods ”. Each experiment was performed in triplicates. Values are given in IL-33 (units) as % of control, which was set as 100% and represent mean ± SD. Experiments were performed 3 times with cells obtained from 3 different donors. *p ≤ 0.05 as compared to untreated control in HACF; **p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACF; §p ≤ 0.05 as compare to untreated control in HACM; §§p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HACM; $p ≤ 0.05 as compared to untreated control in HCASMC; $$p ≤ 0.05 as compared to TNF-α, or IL-1β- or IFN-γ-treated cells in HCASMC.

    Article Snippet: Furthermore, cells were pre-incubated for 30 minutes (min) with the NF-κB inhibitor dimethylfumarate (DMF) (Sigma) at 100 μM before addition of TNF-α, IL-1β or IFN-γ at the indicated concentrations for further 24 h. Additionally, cells were pre-incubated for 30 min with the mitogen-activated protein/extracellular signal-regulated kinase (MEK) inhibitor U0126 (Promega, Madison, WI, USA) at 10 μM or the janus-activated kinase (JAK) 1 and 2 inhibitor JAK inhibitor I (Calbiochem, Merck, Darmstadt, Germany) at 10 μM before addition of TNF-α, IL-1β or IFN-γ for further 24 h. In another set of experiments, HACF, HACM and HCASMC were incubated with rh IL-33 (R & D Systems) at concentrations of 100, 10 or 1 ng/mL or with rh IL-1β at 10 ng/mL (R & D Systems) for time periods between 3 h and 24 h.

    Techniques: Translocation Assay, Expressing, Incubation, Enzyme-linked Immunosorbent Assay

    MIP-1β interfere with monocyte differentiation into DC. Monocytes isolated from an healthy individual were differentiated into DC in presence or not of 200 ng/ml MIP-1β. When indicated, monocytes were treated with the MEK inhibitor (PD) at day 4 and maturation was induced by LPS at day 5. Cells harvested at day 6, were cocultured for 5 days with allogeneic T cells at a ratio of 1/10 DC/T cell in triplicates. IFNγ was measured by CBA in supernatant of cocultures. Mean IFNγ secretion±SD of triplicate wells is shown. * The mean IFNγ secretion stimulated by Mo-DC treated with MIP-1β and LPS is significantly reduced compared to that of MoDC treated with LPS with p

    Journal: PLoS ONE

    Article Title: Th1 Disabled Function in Response to TLR4 Stimulation of Monocyte-Derived DC from Patients Chronically-Infected by Hepatitis C Virus

    doi: 10.1371/journal.pone.0002260

    Figure Lengend Snippet: MIP-1β interfere with monocyte differentiation into DC. Monocytes isolated from an healthy individual were differentiated into DC in presence or not of 200 ng/ml MIP-1β. When indicated, monocytes were treated with the MEK inhibitor (PD) at day 4 and maturation was induced by LPS at day 5. Cells harvested at day 6, were cocultured for 5 days with allogeneic T cells at a ratio of 1/10 DC/T cell in triplicates. IFNγ was measured by CBA in supernatant of cocultures. Mean IFNγ secretion±SD of triplicate wells is shown. * The mean IFNγ secretion stimulated by Mo-DC treated with MIP-1β and LPS is significantly reduced compared to that of MoDC treated with LPS with p

    Article Snippet: In MoDC from HCV patients, TLR4 signaling remains functional since the ability of DC to induce IFNγ secretion by T cells can be restored by treatment with the MEK inhibitor 24 h prior to LPS maturation.

    Techniques: Isolation, Crocin Bleaching Assay

    Effects of MEK or PI3K inhibitors on the production of MMP-3, VEGF and PGE2 by MH7A cells. A . MH7A cells (2 x 10 4 cells/well) were cultured with the MEK inhibitor, PD98059 (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. B . MH7A cells (2 x 10 4 cells/well) were cultured with the PI3K inhibitor, LY294002 (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. Data represent the mean ± SD of three independent experiments. *P

    Journal: SpringerPlus

    Article Title: Differential regulation of c-Met signaling pathways for synovial cell function

    doi: 10.1186/2193-1801-3-554

    Figure Lengend Snippet: Effects of MEK or PI3K inhibitors on the production of MMP-3, VEGF and PGE2 by MH7A cells. A . MH7A cells (2 x 10 4 cells/well) were cultured with the MEK inhibitor, PD98059 (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. B . MH7A cells (2 x 10 4 cells/well) were cultured with the PI3K inhibitor, LY294002 (10 - 60 nM), in the presence or absence of 100 ng/ml TNF-α. After 72 hours of incubation, MMP-3, VEGF and PGE2 production was determined by analysis of supernatants using ELISA. Data represent the mean ± SD of three independent experiments. *P

    Article Snippet: Both the MEK inhibitor, PD98059 and the PI3K inhibitor, LY294002, inhibited MMP-3, VEGF and PGE2 production (Figure ).

    Techniques: Cell Culture, Incubation, Enzyme-linked Immunosorbent Assay

    Detection of pERK signaling of vascular endothelial cells in Vegfr3-Gap43-Venus Tg embryos at E9.5. (A) Flow-cytometry analysis of Vegfr3-Gap43-Venus Tg embryos at E9.5. (B) Western blot analysis of Venus-positive and -negative cells from Tg embryos. (C) Phosphorylated ERK signaling in Venus-positive cells cultured in the presence or absence of the MEK inhibitor. (D) Percentage of pERK-positive cells in Venus-positive cells.

    Journal: PLoS ONE

    Article Title: Generating Vegfr3 reporter transgenic mouse expressing membrane-tagged Venus for visualization of VEGFR3 expression in vascular and lymphatic endothelial cells

    doi: 10.1371/journal.pone.0210060

    Figure Lengend Snippet: Detection of pERK signaling of vascular endothelial cells in Vegfr3-Gap43-Venus Tg embryos at E9.5. (A) Flow-cytometry analysis of Vegfr3-Gap43-Venus Tg embryos at E9.5. (B) Western blot analysis of Venus-positive and -negative cells from Tg embryos. (C) Phosphorylated ERK signaling in Venus-positive cells cultured in the presence or absence of the MEK inhibitor. (D) Percentage of pERK-positive cells in Venus-positive cells.

    Article Snippet: Flow-cytometry analysis with the pERK antibody Venus-positive and–negative cells from Vegfr3-Gap43-Venus BAC Tg embryos at E9.5 and were cultured in the presence of 5 μM MEK inhibitor (PD0325901; Wako, Osaka, Japan) for 30 min in a CO2 incubator.

    Techniques: Flow Cytometry, Cytometry, Western Blot, Cell Culture

    DAZAP1 regulates cell proliferation (a) Stable over-expression of DAZAP1 in non-small cell lung carcinoma NCI-H157 cells as detected by western blots. Control cells are stably transfected with vector only. (b) The expression of DAZAP1 inhibited cell proliferation as judged by colony formation assay with H157 cells. A representative picture from the triplicate experiments, the means and s.d. were plotted. (c) DAZAP1 inhibited the anchorage independent cell growth of H157 as judged by soft agar assays. A representative picture from the triplicates and the quantification were shown. The box plot was drawn using 25 and 75 quartiles in Sigma Plot with whiskers indicating max and min value and dots indicating outliers. P values were determined using paired t-test, *=p ≤ 5.0×10 −5 . (d) DAZAP1 inhibited cell migration as judged by wound healing assay. The picture of a representative plate (left panel) and the quantification of percent of gap closure were shown (right panel). Data from three independent assays were performed error bar indicate s.d. (e) The quantification of cell cycle stages (G1, S and G2/M phases) in DAZAP1 over-expressed or control cells. The cell cycle stages were analyzed by flow-cytometry measurement of DNA content. Data from two independent flow analysis, error bar indicate s.d. (f) Expression of endogenous DAZAP1 protein during cell cycle as detected by western blots. Tubulin and canonical cell cycle markers (cyclin A1 and B1) were also probed as controls. (g) An integrated model for DAZAP1 function and the regulation of its activity through the MEK/Erk pathway.

    Journal: Nature communications

    Article Title: The splicing activator DAZAP1 integrates splicing control into MEK/Erk regulated cell proliferation and migration

    doi: 10.1038/ncomms4078

    Figure Lengend Snippet: DAZAP1 regulates cell proliferation (a) Stable over-expression of DAZAP1 in non-small cell lung carcinoma NCI-H157 cells as detected by western blots. Control cells are stably transfected with vector only. (b) The expression of DAZAP1 inhibited cell proliferation as judged by colony formation assay with H157 cells. A representative picture from the triplicate experiments, the means and s.d. were plotted. (c) DAZAP1 inhibited the anchorage independent cell growth of H157 as judged by soft agar assays. A representative picture from the triplicates and the quantification were shown. The box plot was drawn using 25 and 75 quartiles in Sigma Plot with whiskers indicating max and min value and dots indicating outliers. P values were determined using paired t-test, *=p ≤ 5.0×10 −5 . (d) DAZAP1 inhibited cell migration as judged by wound healing assay. The picture of a representative plate (left panel) and the quantification of percent of gap closure were shown (right panel). Data from three independent assays were performed error bar indicate s.d. (e) The quantification of cell cycle stages (G1, S and G2/M phases) in DAZAP1 over-expressed or control cells. The cell cycle stages were analyzed by flow-cytometry measurement of DNA content. Data from two independent flow analysis, error bar indicate s.d. (f) Expression of endogenous DAZAP1 protein during cell cycle as detected by western blots. Tubulin and canonical cell cycle markers (cyclin A1 and B1) were also probed as controls. (g) An integrated model for DAZAP1 function and the regulation of its activity through the MEK/Erk pathway.

    Article Snippet: Addition of MEK inhibitor (U0126, Sigma) was followed 2 h post transfection and cells were collected after 16–18h-post treatment for protein and splicing analysis.

    Techniques: Over Expression, Western Blot, Stable Transfection, Transfection, Plasmid Preparation, Expressing, Colony Assay, Migration, Wound Healing Assay, Flow Cytometry, Cytometry, Activity Assay

    DAZAP1 is regulated by Erk1/2 catalyzed threonine phosphorylation in the CTD ( a ). A series of truncated CTDs of DAZAP1 were tethered to the exon of a splicing reporter to test which regions are critical for its activity. The full length DAZAP1 was used as positive control, and the two Erk1/2 phosphorylation sites (Thr269 and Thr315) are marked by red triangle. The experimental conditions were identical to Fig. 3b . The mean PSI values and s.d. are shown below a representative gel. P values were determined with paired t-test by comparing to vector-only control. In all panels, #: p ≤0.05; *= p ≤0.01; **= p ≤0.002 ; *** = p ≤0.0008; ns= not significant. ( b ) A schematic model showing that DAZAP1 activity is mediated by MEK/Erk pathway (left panel). The tethering experiments similar to panel a were conducted in cells treated with MEK/Erk activator (PMA) or inhibitor (U0126). ( c ). Phosphorylation of DAZAP1 was inhibited by two specific MEK inhibitors as judged by western blot with Phos-tag labeling. The phosphorylated forms of DAZAP1 were indicated with arrows. (d) Mutation of the two DAZAP1 threonine sites phosphorylated by Erk1/2 abolished DAZAP1 activity as judged by tethering experiments. The experiments were conducted in triplicates and the mean and s.d. of PSI values are plotted below a representative gel. The western blot was shown on the right to confirm equal expression between wild type and mutated DAZAP1. The band indicated by the asterisk is likely a degradation product of MS2-DAZAP1. (e) Inhibition of MEK/Erk by U0126 affected the splicing of endogenous DAZAP1 targets FIP1L1 and MELK. A representative gel from duplicated experiments and the relative changes of PSI were plotted with ranges as error bars. ( f ) Inhibition of MEK/Erk pathway affected nuclear/cytoplasmic translocation of DAZAP1. The cells were transfected with tagged DAZAP1 and hnRNP A1 and detected by corresponding antibodies. Bar, 5 μm. The percents of cells with DAZAP1 localized in nucleus (N), cytoplasm (C) or both compartments (N+C) were counted and plotted in the right panel (n=80 for each sample in two independent experiments). ( g ) Inhibition of MEK/Erk pathway affected nuclear/cytoplasmic translocation of endogenous DAZAP1. Scale bar, 2.5 μm.

    Journal: Nature communications

    Article Title: The splicing activator DAZAP1 integrates splicing control into MEK/Erk regulated cell proliferation and migration

    doi: 10.1038/ncomms4078

    Figure Lengend Snippet: DAZAP1 is regulated by Erk1/2 catalyzed threonine phosphorylation in the CTD ( a ). A series of truncated CTDs of DAZAP1 were tethered to the exon of a splicing reporter to test which regions are critical for its activity. The full length DAZAP1 was used as positive control, and the two Erk1/2 phosphorylation sites (Thr269 and Thr315) are marked by red triangle. The experimental conditions were identical to Fig. 3b . The mean PSI values and s.d. are shown below a representative gel. P values were determined with paired t-test by comparing to vector-only control. In all panels, #: p ≤0.05; *= p ≤0.01; **= p ≤0.002 ; *** = p ≤0.0008; ns= not significant. ( b ) A schematic model showing that DAZAP1 activity is mediated by MEK/Erk pathway (left panel). The tethering experiments similar to panel a were conducted in cells treated with MEK/Erk activator (PMA) or inhibitor (U0126). ( c ). Phosphorylation of DAZAP1 was inhibited by two specific MEK inhibitors as judged by western blot with Phos-tag labeling. The phosphorylated forms of DAZAP1 were indicated with arrows. (d) Mutation of the two DAZAP1 threonine sites phosphorylated by Erk1/2 abolished DAZAP1 activity as judged by tethering experiments. The experiments were conducted in triplicates and the mean and s.d. of PSI values are plotted below a representative gel. The western blot was shown on the right to confirm equal expression between wild type and mutated DAZAP1. The band indicated by the asterisk is likely a degradation product of MS2-DAZAP1. (e) Inhibition of MEK/Erk by U0126 affected the splicing of endogenous DAZAP1 targets FIP1L1 and MELK. A representative gel from duplicated experiments and the relative changes of PSI were plotted with ranges as error bars. ( f ) Inhibition of MEK/Erk pathway affected nuclear/cytoplasmic translocation of DAZAP1. The cells were transfected with tagged DAZAP1 and hnRNP A1 and detected by corresponding antibodies. Bar, 5 μm. The percents of cells with DAZAP1 localized in nucleus (N), cytoplasm (C) or both compartments (N+C) were counted and plotted in the right panel (n=80 for each sample in two independent experiments). ( g ) Inhibition of MEK/Erk pathway affected nuclear/cytoplasmic translocation of endogenous DAZAP1. Scale bar, 2.5 μm.

    Article Snippet: Addition of MEK inhibitor (U0126, Sigma) was followed 2 h post transfection and cells were collected after 16–18h-post treatment for protein and splicing analysis.

    Techniques: Activity Assay, Positive Control, Plasmid Preparation, Western Blot, Labeling, Mutagenesis, Expressing, Inhibition, Translocation Assay, Transfection

    Rationale for Targeting Both the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR Pathways for Suppressing Cancer Growth A: The Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways are both activated by upstream receptor ligation and frequently co-regulate many downstream targets in parallel. Thus for effective elimination of many cancers or prevention of aging, it may be necessary to target both signaling pathways. Activation of these pathways could also result in increased transcription of many genes that promote cellular growth and malignant transformation. B. Inhibition of mTOR can result in the induction of autophagy, which is a very important mechanism of cell death, especially in solid tumors. C. As described previously, both the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways regulate the activity of apoptotic proteins by post-translational mechanisms. Targeting this pathway may also contribute to the induction of apoptosis. Signaling molecules promoting phosphorylation events are indicated in green. Stimulatory signaling events are indicted in green lines with a green arrow before the target of the phophorylation. Small molecule inhibitors are indicated in red. Inhibitory phosphorylation events are indicated in red lines with a block on the end before the target of the inhibition. Inhibitory signaling or proapoptotic molecules or inactivated molecules are indicated in yellow. A growth factor and a growth factor receptor are indicated in purple. Active transcription factors are indicated in purple diamonds. Inactivated transcription factors are indicated in yellow diamonds.

    Journal: Oncotarget

    Article Title: Ras/Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR Inhibitors: Rationale and Importance to Inhibiting These Pathways in Human Health

    doi:

    Figure Lengend Snippet: Rationale for Targeting Both the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR Pathways for Suppressing Cancer Growth A: The Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways are both activated by upstream receptor ligation and frequently co-regulate many downstream targets in parallel. Thus for effective elimination of many cancers or prevention of aging, it may be necessary to target both signaling pathways. Activation of these pathways could also result in increased transcription of many genes that promote cellular growth and malignant transformation. B. Inhibition of mTOR can result in the induction of autophagy, which is a very important mechanism of cell death, especially in solid tumors. C. As described previously, both the Ras/Raf/MEK/ERK and Ras/PI3K/PTEN/Akt/mTOR pathways regulate the activity of apoptotic proteins by post-translational mechanisms. Targeting this pathway may also contribute to the induction of apoptosis. Signaling molecules promoting phosphorylation events are indicated in green. Stimulatory signaling events are indicted in green lines with a green arrow before the target of the phophorylation. Small molecule inhibitors are indicated in red. Inhibitory phosphorylation events are indicated in red lines with a block on the end before the target of the inhibition. Inhibitory signaling or proapoptotic molecules or inactivated molecules are indicated in yellow. A growth factor and a growth factor receptor are indicated in purple. Active transcription factors are indicated in purple diamonds. Inactivated transcription factors are indicated in yellow diamonds.

    Article Snippet: In a study with NSCLC cells which constitutively-expressed activated MEK/ERK, no increase in paclitaxel-induced apoptosis was observed when the cells were treated with a MEK inhibitor [ ].

    Techniques: Ligation, Activation Assay, Transformation Assay, Inhibition, Activity Assay, Blocking Assay

    Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 (MEK inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories

    Journal: Nature Communications

    Article Title: Dynamics of lineage commitment revealed by single-cell transcriptomics of differentiating embryonic stem cells

    doi: 10.1038/s41467-017-01076-4

    Figure Lengend Snippet: Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 (MEK inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories

    Article Snippet: Cell culture All cell lines were grown routinely in modified 2i medium plus LIF (2i/L): DMEM/F12 (Life technologies) supplemented with 0.5x N2 supplement, 0.5x B27 supplement, 0.5mM L -glutamine (Gibco), 20 µg/ml human insulin (Sigma-Aldrich), 1 × 100U/ml penicillin/streptomycin (Gibco), 0.5x MEM Non-Essential Amino Acids (Gibco), 0.1 mM 2-Mercaptoethanol (Sigma-Aldrich), 1 µM MEK inhibitor (PD0325901, Stemgent), 3 µM GSK3 inhibitor (CHIR99021, Stemgent), 1000 U/ml mouse LIF (ESGRO).

    Techniques: Expressing, Staining, Flow Cytometry, Cytometry, Incubation, Activation Assay

    MEK inhibitor inhibits CVB3-mediated RasGAP cleavage. HeLa cells were pretreated for 30 min with various concentrations of U0126 and then were infected by CVB3 for 1 h. At 9 h after addition of CVB3, HeLa cells were harvested and Western blot analysis was performed using an antibody that recognizes RasGAP. Results were quantitated by densitometric analysis using National Institutes of Health Image, version 1.61, and normalized to the control level (sham-infected cells without U0126), which was arbitrarily set to 1.0. Values are means ± standard errors ( n = 3).

    Journal: Journal of Virology

    Article Title: Coxsackievirus B3 Replication Is Reduced by Inhibition of the Extracellular Signal-Regulated Kinase (ERK) Signaling Pathway

    doi: 10.1128/JVI.76.7.3365-3373.2002

    Figure Lengend Snippet: MEK inhibitor inhibits CVB3-mediated RasGAP cleavage. HeLa cells were pretreated for 30 min with various concentrations of U0126 and then were infected by CVB3 for 1 h. At 9 h after addition of CVB3, HeLa cells were harvested and Western blot analysis was performed using an antibody that recognizes RasGAP. Results were quantitated by densitometric analysis using National Institutes of Health Image, version 1.61, and normalized to the control level (sham-infected cells without U0126), which was arbitrarily set to 1.0. Values are means ± standard errors ( n = 3).

    Article Snippet: U0126, the MEK inhibitor, was purchased from Promega.

    Techniques: Infection, Western Blot

    The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and U0126 MEK inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P

    Journal: Biology of Reproduction

    Article Title: Dose-dependent functions of fibroblast growth factor 9 regulate the fate of murine XY primordial germ cells

    doi: 10.1095/biolreprod.116.143941

    Figure Lengend Snippet: The effect of ERK1/2 signaling activity on proliferation of XY PGCs cultured with high FGF9. A ) Five thousand XY PGCs isolated from 12.5 dpc male gonads were incubated with high FGF9 for 2 hr to promote phosphorylation of ERK1/2. When the cells were incubated with both high FGF9 and U0126 MEK inhibitor (10 μM) for 30 min or 1 hr, ERK1/2 activity induced by high FGF9 was completely suppressed. The levels of protein bands were quantified by densitometry and normalized to the β-ACTIN levels. The relative optical densities of phosphorylated protein levels were calculated based on the level in the high FGF9 treatment group set as 1.00. B) To determine the relationship between the ERK1/2 signaling pathway and PGC proliferation when stimulated by high FGF9 treatment, XY PGCs isolated at 12.5 dpc were cultured with high FGF9 and U0126 inhibitor (1, 10 μM) for 2 days and then subjected to the EdU cell proliferation assay. Cells treated with high FGF9 alone significantly increased cell proliferation ( P

    Article Snippet: In some experiments, PGCs were cultured with p38 inhibitors (BIRB796; Cayman Chemical Company, Ann Arbor, MI, SB202190; Sigma-Aldrich) at 10 μM or MEK inhibitor (U0126; Cayman Chemical Company) at 1 or 10 μM.

    Techniques: Activity Assay, Cell Culture, Isolation, Incubation, Pyrolysis Gas Chromatography, Proliferation Assay

    MEK inhibitor suppressed spheroid formation in ID8-KRAS cells. ID8-KRAS-GFP cells (2 × 10 5 ) were not treated (0 nM) or treated with trametinib (1, 10, 100 nM) for 48 h in 2D or 3D culture. a Representative microscopic image of cancer cells under 2D and 3D conditions at 100× magnification. b-c After the treatment, cells were exposed to EdU (10 μM) for 2 h and fixed. EdU was labeled with Alexa Fluor 647. EdU uptake was analyzed ( b ) and the ratio of EdU-positive to EdU-negative cells was measured by flow cytometry ( c ). Experiments were repeated at least thrice. The values shown represent mean ± SEM (* p

    Journal: BMC Cancer

    Article Title: The oncogene KRAS promotes cancer cell dissemination by stabilizing spheroid formation via the MEK pathway

    doi: 10.1186/s12885-018-4922-4

    Figure Lengend Snippet: MEK inhibitor suppressed spheroid formation in ID8-KRAS cells. ID8-KRAS-GFP cells (2 × 10 5 ) were not treated (0 nM) or treated with trametinib (1, 10, 100 nM) for 48 h in 2D or 3D culture. a Representative microscopic image of cancer cells under 2D and 3D conditions at 100× magnification. b-c After the treatment, cells were exposed to EdU (10 μM) for 2 h and fixed. EdU was labeled with Alexa Fluor 647. EdU uptake was analyzed ( b ) and the ratio of EdU-positive to EdU-negative cells was measured by flow cytometry ( c ). Experiments were repeated at least thrice. The values shown represent mean ± SEM (* p

    Article Snippet: Our assessment of the therapeutic potential of a MEK-ERK inhibitor on spheroid formation revealed that the MEK inhibitor decreased spheroid formation by ID8-KRAS cells in 3D culture, but not in 2D culture.

    Techniques: Labeling, Flow Cytometry, Cytometry

    MEK inhibitor suppressed spheroid formation in ID8-KRAS mice in vivo . Mice were injected intraperitoneally with ID8-KRAS-GFP cells (1 × 10 6 ) and treated with the MEK inhibitor trametinib or the vehicle (control). Mice were sacrificed when their body weight exceeded 23 g after inoculation. a Amount of ascites was assessed at the time of sacrifice. A statistical analysis was performed with the Student’s t-test (* p

    Journal: BMC Cancer

    Article Title: The oncogene KRAS promotes cancer cell dissemination by stabilizing spheroid formation via the MEK pathway

    doi: 10.1186/s12885-018-4922-4

    Figure Lengend Snippet: MEK inhibitor suppressed spheroid formation in ID8-KRAS mice in vivo . Mice were injected intraperitoneally with ID8-KRAS-GFP cells (1 × 10 6 ) and treated with the MEK inhibitor trametinib or the vehicle (control). Mice were sacrificed when their body weight exceeded 23 g after inoculation. a Amount of ascites was assessed at the time of sacrifice. A statistical analysis was performed with the Student’s t-test (* p

    Article Snippet: Our assessment of the therapeutic potential of a MEK-ERK inhibitor on spheroid formation revealed that the MEK inhibitor decreased spheroid formation by ID8-KRAS cells in 3D culture, but not in 2D culture.

    Techniques: Mouse Assay, In Vivo, Injection

    Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 (MEK inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories

    Journal: Nature Communications

    Article Title: Dynamics of lineage commitment revealed by single-cell transcriptomics of differentiating embryonic stem cells

    doi: 10.1038/s41467-017-01076-4

    Figure Lengend Snippet: Susceptibility to signaling inputs was highly dynamic around the exit from pluripotency. a – c Fractions of cells classified as XEN-like, ectoderm-like, double positive and double negative after 96 h, based on CD24 and PDGFRA expression. Expression of the two markers was measured by antibody staining and flow cytometry. a Cells were pulsed with 0.25 µM RA for x h (pulse) and subsequently differentiated in basal medium (N2B27) complemented with an RA receptor antagonist. b Cells were incubated with 0.25 µM RA for x h (pulse) after which 0.5 µM PD0325901 (MEK inhibitor) was added for the remainder of the time course. c Cells were first incubated with basal medium (N2B27) for x h (delay) and then exposed to 0.25 µM RA for the remainder of the time course. d Schematic representation of a minimal gene regulatory network that can model a lineage decision 32 . Pointy arrows indicate (auto-)activation; blunted arrows indicate repression. E and X represent expression of ectoderm-like and XEN-like transcriptional programs, respectively. P stands for the pluripotency network. RA increases the auto-activation of the XEN program. e Results of the stochastic simulations of the network shown in d . The relative frequency of XEN-like cells after 96 h is shown vs. the length of an RA pulse (solid lines) or the length of the delay before RA exposure is started (dashed lines). In all cases the pluripotency network was turned off after 12 h. Simulations were run with different amounts of gene expression noise (D: noise power / time, see Methods). See Supplementary Fig. 8b for exemplary trajectories

    Article Snippet: For the differentiation under perturbation of various signaling pathways (Supplementary Fig. ) we used the MEK inhibitor PD0325901 (Stemgent, standard concentration 1 µM or dilutions thereof), GSK3 inhibitor CHIR99021 (Stemgent, standard concentration 3 µM or dilutions thereof) or mouse LIF (ESGRO, 1000 U/ml).

    Techniques: Expressing, Staining, Flow Cytometry, Cytometry, Incubation, Activation Assay