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Identify the downstream targets of Stat3 by using an engineered mouse ES cell line treated with GCSF and LIF plus PD0325901

ABSTRACT: To identify downstream targets of Jak/Stat3 pathways without being distracted by differentiation signalings from MEK/ERK pathway, we exploited a engineered B6 cells, which stably stably expressing a chimeric receptor (GRgp-Y118F). The chimeric receptor can induce the phosphorylation of Stat3 by GCSF without activating the MEK/ERK pathway. To mimic the effect of GCSF, the chimeric B6 cells were also treated with LIF plus a selective MEK chemical inhibitor, PD0325901, to induce LIF/Jak/Stat3 but MEK/ERK pathways.

ORGANISM(S): Mus musculus

PROVIDER: GSE38719 | GEO | 2012/06/14

SECONDARY ACCESSION(S): PRJNA168497

REPOSITORIES: GEO

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Identify the downstream targets of Stat3 by using an engineered mouse ES cell line treated with GCSF and LIF plus PD0325901

Project description:To identify downstream targets of Jak/Stat3 pathways without being distracted by differentiation signalings from MEK/ERK pathway, we exploited a engineered B6 cells, which stably stably expressing a chimeric receptor (GRgp-Y118F). The chimeric receptor can induce the phosphorylation of Stat3 by GCSF without activating the MEK/ERK pathway. To mimic the effect of GCSF, the chimeric B6 cells were also treated with LIF plus a selective MEK chemical inhibitor, PD0325901, to induce LIF/Jak/Stat3 but MEK/ERK pathways. mESCs starved in serum free growth medium for 6hrs were treated with GCSF or with LIF plus PD0325901 for 1hr, after which total RNA was extracted for analysis.

2012-06-13 | E-GEOD-38719 | biostudies-arrayexpress

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Project description:FGF-MEK-ERK pathway has been implicated in diverse contexts in development. The pathway involves many autoinhibitory loops, resulting in pulse-like activity patterns. Here, we modulated the ERK activity during cardiac differentiation at the lateral plate mesoderm stage by PD0325901, a potent inhibitor of MEK, which is an upstream effector of ERK. We collected the resulting cell population, both in 2D and 3D, after cardiac specification to analyse how the ERK inhibition during induction affects the cell fate choice.

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Project description:The role of FGF-MEK-ERK signalling pathway during embryonic heart development has not been fully elucidated. Here, we inhibited the pathway for 1 day using PD0325901, a MEK inhibitor, at the lateral plate mesoderm stage during cardiac differentiation of human embryonic stem cells. Cells were collected on day 2 (before PD0325901 administration), day 3 and day 8 to determine the effect of a transient FGF-MEK-ERK pathway modulation on the cardiac cell fate choice.

2023-06-28 | E-MTAB-12619 | biostudies-arrayexpress

Genome-wide analysis of gene expression in LIF and LIF plus 2I in C57BL/6 mouse embryonic stem cells (B6 mESCs)

Project description:Analysis of genes induced by 2I condition 2i contains glycogen synthase kinase 3 (GSK3) and mitogen-activated protein kinase kinase (MEK) inhibitors: 3uM Chir99021 and 1uM PD0325901 Total RNA obtained from B6 mESCs treated with LIF or LIF/2I for 12 hours.

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Combined MEK and JAK/STAT3 pathway inhibition effectively decreases medulloblastoma tumor progression.

Project description:RNA sequencing was carried out to identify the signaling pathways and hallmarks that change following MEK and/or JAK/STAT3 pathway inhibition.

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12h pathway stimulation / inhibition experiment in epiblast stem cells

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Transcription profiling of human cell lines in culture (n=12) and murine xenografts (n=2) were treated with the MEK inhibitor PD0325901 or vehicle alone as controlPaired MEK inhibited and control

Project description:This study used microarray expression analysis to identify global changes in transcript alteration in response to MEK inhibition. Genes under ERK control were identified in a panel of V600E BRAF and RTK-activated tumor cells and xenografts, using short-term inhibition of ERK activity using the MEK inhibitor PD0325901 (Pfizer). Experiment Overall Design: Cell lines growing in culture (n=12) and murine xenografts (n=2) were treated with the MEK inhibitor PD0325901 or vehicle alone as control. Paired analysis of MEK inhibited to control samples was performed for two groups of tumor cells, V600E BRAF and RTK.

2009-02-21 | E-GEOD-10087 | biostudies-arrayexpress

DNMTs Play an Important Role in Maintaining the Pluripotency of Leukemia Inhibitory Factor Dependent Embryonic Stem Cells

Project description:Naïve pluripotency can be maintained in 2i/LIF supplements, which primarily affect canonical WNT, FGF/ERK, and JAK/STAT3 signaling. However, whether one of these tripartite supplements alone is sufficient to maintain naïve self-renewal remain unclear. Here we show that LIF alone medium is sufficient for adaptation of 2i/L-ESCs to ESCs with hypermethylated state (L-ESCs). Global transcriptomic analysis show that L-ESCs are close to 2i/L-ESCs and in a stable state between naïve and primed pluripotency. Notably, our results demonstrate that DNA methyltransferases (DNMTs) play an important role for LIF-dependent mouse ESCs adaptation and self-renew. LIF-dependent ESCs adaptation efficiency is significantly increased in serum treatment and reduced in Dnmt3a or Dnmt3l knockout ESCs. Importantly, unlike epiblast stem cells, L-ESCs contribute to somatic tissues and germ cells in chimaeras. L-ESCs cultured in such a simple conditions in this study would provide a more conducive platform to clarify the molecular mechanism of ESCs in vitro culture.

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