CD71high population represents primitive erythroblasts derived from mouse embryonic stem cells

Chao, Robert; Gong, Xueping; Wang, Libo; Wang, Pengxiang; Wang, Yuan

doi:10.1016/j.scr.2014.11.002

Cited by 12 publications

(12 citation statements)

References 28 publications

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“…2H). We further noticed a slight but significant increase in CD71 high cells upon AP-1 inhibition, which was reported to represent a population of primitive erythroblasts (Chao et al, 2015). To test whether floating cells derived from dnFOS-induced cultures have the potential to form normal colony when compared with untreated cultures, we performed colony-forming assays in the absence of DOX.…”

Section: Resultsmentioning

confidence: 55%

Cooperative binding of AP-1 and TEAD4 modulates the balance between vascular smooth muscle and hemogenic cell fate

et al. 2016

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The transmission of extracellular signals into the nucleus involves inducible transcription factors, but how different signalling pathways act in a cell type-specific fashion is poorly understood. Here, we studied the regulatory role of the AP-1 transcription factor family in blood development using embryonic stem cell differentiation coupled with genome-wide transcription factor binding and gene expression analyses. AP-1 factors respond to MAP kinase signalling and comprise dimers of FOS, ATF and JUN proteins. To examine genes regulated by AP-1 and to examine how it interacts with other inducible transcription factors, we abrogated its global DNA-binding activity using a dominant-negative FOS peptide. We show that FOS and JUN bind to and activate a specific set of vascular genes and that AP-1 inhibition shifts the balance between smooth muscle and hematopoietic differentiation towards blood. Furthermore, AP-1 is required for de novo binding of TEAD4, a transcription factor connected to Hippo signalling. Our bottom-up approach demonstrates that AP-1- and TEAD4-associated cis-regulatory elements form hubs for multiple signalling-responsive transcription factors and define the cistrome that regulates vascular and hematopoietic development by extrinsic signals.

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Section: Resultsmentioning

confidence: 55%

Cooperative binding of AP-1 and TEAD4 modulates the balance between vascular smooth muscle and hemogenic cell fate

et al. 2016

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“…This problem is commonly encountered in live imaging studies of dynamic cell processes and so, to overcome this, we stratified erythroblasts obtained from these cultures using directly-conjugated antibodies (Eilken et al, 2009) that recognize CD71 and Ter119, known surface markers of erythropoiesis whose levels change in a predictable manner during erythroid differentiation (Socolovsky et al, 2001; Zhang et al, 2003, Fraser et al, 2006, Chao et al, 2015). Although erythroid cells can be separated in various stages of differentiation using fluorescently activated cell sorting (FACS), we observed reduced viability of sorted cells in prolonged imaging experiments, potentially caused by an extended period of sorting-associated stress.…”

Section: Resultsmentioning

confidence: 99%

On-microscope staging of live cells reveals changes in the dynamics of transcriptional bursting during differentiation

Jeziorska

Tunnacliffe

Brown

et al. 2021

Preprint

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Determining the mechanisms by which genes are switched on and off during development and differentiation is a key aim of current biomedical research. Gene transcription has been widely observed to occur in a discontinuous fashion, with short bursts of activity interspersed with longer periods of inactivity. It is currently not known if or how this dynamic behaviour changes as mammalian cells differentiate. To investigate this, using a newly developed on-microscope analysis, we monitored mouse α-globin transcription in live cells throughout sequential stages of erythropoiesis. We find that changes in the overall levels of α-globin transcription are most closely associated with changes in the fraction of time a gene spends in the active transcriptional state. We identify differences in the patterns of transcriptional bursting throughout differentiation, with maximal transcriptional activity occurring in the mid-phase of differentiation. Early in differentiation, we observe increased fluctuation in the patterns of transcriptional activity whereas at the peak of gene expression, in early and intermediate erythroblasts, transcription appears to be relatively stable and efficient. Later during differentiation as α-globin expression declines, we again observed more variability in transcription within individual cells. We propose that the observed changes in transcriptional behaviour may reflect changes in the stability of enhancer-promoter interactions and the formation of active transcriptional compartments as gene expression is turned on and subsequently declines at sequential stages of differentiation.

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“…Flow cytometry analyses of mouse foetal erythroblasts with CD71 and Ter119 were performed as previously described 11,42 . For enucleation analysis, cells were stained with 10 µg/mL Hoechst 33342 (Sigma–Aldrich), together with an APC‐Ter119 antibody, for 30 minutes at 4°C.…”

Section: Methodsmentioning

confidence: 99%

CAP1, a target of miR‐144/451, negatively regulates erythroid differentiation and enucleation

Huang

Chao

Zhang

et al. 2021

J Cellular Molecular Medi

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The exact molecular mechanism underlying erythroblast enucleation has been a fundamental biological question for decades. In this study, we found that miR‐144/451 critically regulated erythroid differentiation and enucleation. We further identified CAP1, a G‐actin‐binding protein, as a direct target of miR‐144/451 in these processes. During terminal erythropoiesis, CAP1 expression declines along with gradually increased miR‐144/451 levels. Enforced CAP1 up‐regulation inhibits the formation of contractile actin rings in erythroblasts and prevents their terminal differentiation and enucleation. Our findings reveal a negative regulatory role of CAP1 in miR‐144/451‐mediated erythropoiesis and thus shed light on how microRNAs fine‐tune terminal erythroid development through regulating actin dynamics.

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CD71high population represents primitive erythroblasts derived from mouse embryonic stem cells

Cited by 12 publications

References 28 publications

Cooperative binding of AP-1 and TEAD4 modulates the balance between vascular smooth muscle and hemogenic cell fate

Cooperative binding of AP-1 and TEAD4 modulates the balance between vascular smooth muscle and hemogenic cell fate

On-microscope staging of live cells reveals changes in the dynamics of transcriptional bursting during differentiation

CAP1, a target of miR‐144/451, negatively regulates erythroid differentiation and enucleation

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