Project description:ChIP-seq assay was performed with anti-p45 antibody using in vitro cultured primary megakaryocytes to identify direct p45 target genes in megakaryocytes. This analysis revealed genes that regulate platelet function, which were not known previously to be p45-regulated. ChIP was performed using a day-3 primary culture of megakaryocytes from E13.5 fetal livers of wild-type mice with an anti-p45 antibody.

2013-11-01 | E-GEOD-42249 | biostudies-arrayexpress

Comparison of gene expression profiles of p45-/- primary megakaryocytes and WT primary megakaryocytes

Project description:Whole livers were collected from mouse fetuses at embryonic day 13.5 (E13.5), and single-cell suspensions were prepared by successive passage through 25-gauge needles. Fetal liver cells were maintained in RPMI1640 (Wako, Osaka, Japan) supplemented with 20% charcoal-stripped fetal bovine serum (FBS), 100 U/ml penicillin, 100µg/ml streptomycin, and 50ng/ml recombinant human thrombopoietin (TPO) (generously provided by Kyowa Hakko Kirin Co. Ltd.). Megakaryocytes were harvested for RNA purification from a day-3 culture. CD41+ cells were enriched using biotinylated anti-CD41 antibody (Serotec; clone MWReg30) and streptavidin-coupled Dynabeads (Dynal Biotech ASA) from a culture of E13.5 fetal liver cells of WT and p45–/– mice. Total RNA was purified from the sorted CD41+ cells. Gene expression was measured in primary megakaryocytes cultured from p45-/- fetal livers and wild type fetal livers at E13.5. Three independent experiments were performed.

2013-08-07 | E-GEOD-35645 | biostudies-arrayexpress

Mouse primary megakaryocytes, wild type & NF-E2p45-deficient

2010-03-22 | GSE15581 | GEO

Mus musculus

Project description:NF-E2, FLI1 and RUNX1 collaborate at areas of dynamic chromatin to activate transcription in mature mouse megakaryocytes

| PRJNA179147 | ENA

Mouse primary megakaryocytes, wild type & NF-E2p45-deficient

Project description:To identify p45 target genes, we conducted gene expression profiling with p45-null megakaryocytes cultured from E14.5 fetal liver. Many genes encoding membrane proteins and enzymes related to platelet function, including Txas, Glycoprotein 6 (Gp6) and Selectin P (Selp), were repressed in the absence of p45. Considering the similar DNA binding specificity of p45 and Nrf2 in vitro, we expected p45 to activate cytoprotective genes that are established Nrf2 targets. However, the expression of numerous detoxifying enzymes and stress-responsive genes, including NAD(P)H:quinone oxidoreductase (Nqo1), were increased in the absence of p45. To obtain wild type and p45-null fetal livers, p45+/- mice were crossed. Whole livers were recovered from mouse fetuses at E14.5 and single cell suspensions were prepared by successive passage through 25-gauge needles. Fetal liver cells were maintained in RPMI1640 (Wako) supplemented with 20% charcoal-stripped fetal bovine serum, 50 units/ml penicillin, 50 Âµg/ml streptomycin, and 50 ng/ml of recombinant human thrombopoietin. CD41+ cells were selected from a 4 day primary culture of E14.5 fetal liver cells using a MACS magnetic system (Miltenyi Biotec). The fetal liver culture with TPO described above was incubated with FITC-conjugated anti-CD41 antibody (BD Pharmingen, clone MWReg30) followed by incubation with anti-FITC microbeads. Subsequently, the microbeads were selected magnetically through MACS large cell columns (Miltenyi Biotec). Total RNA was extracted from the CD41+ cells using Isogen (Nippon Gene).

2010-03-22 | E-GEOD-15581 | biostudies-arrayexpress

Mus musculus

Project description:Comparison of gene expression profiles of miR-142-/- primary megakaryocytes and WT primary megakaryocytes

| PRJNA227016 | ENA

Single cell RNA-seq of mouse megakaryocytes from wild type and Nf-e2 knock out mice

Project description:Single cell RNA-seq of mouse megakaryocytes from wild type and Nf-e2 knock out mice

| PRJNA516873 | ENA