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Effect of depletion of Hand1 and Foxf1 on gene expression during CM differentiation of E14 embryonic stem cells II

ABSTRACT: To investigate the function of Hand1 and Foxf1 in the regulation of Cardiac differentiation, we established E14 cell lines in which each target gene has been knocked out by sgRNA. We then performed gene expression profiling analysis using data obtained from RNA-seq of these cell lines at different time points during the CM differentiation process.

ORGANISM(S): Mus musculus

PROVIDER: GSE245711 | GEO | 2025/12/08

REPOSITORIES: GEO

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Effect of depletion of Hand1 and Foxf1 on gene expression during CM differentiation of E14 embryonic stem cells I

Project description:To investigate the function of Hand1 and Foxf1 in the regulation of Cardiac differentiation, we established E14 cell lines in which each target gene has been knocked out by sgRNA.

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Effect of depletion of Hand1 and Foxf1 on gene expression during CM differentiation of E14 embryonic stem cells II

Project description:Effect of depletion of Hand1 and Foxf1 on gene expression during CM differentiation of E14 embryonic stem cells II

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Effect of depletion of Hand1 and Foxf1 on gene expression during CM differentiation of E14 embryonic stem cells I

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HAND1 ChIP-seq at day 3 of cardiac differentiation from human embryonic stem cells

Project description:The aim was to identify HAND1 binding sites at low and high HAND1 levels at an early stage of hESC cardiac differentiation. We generated a HAND1-AM-Tag (Active Motif) knock-in to enable sensitive and specific ChIP-seq analysis. The hESCs were differentiated in 3D embryoid bodies with BMP4, Activin A and CHIR. To increase HAND1 levels, we treated one population with SB431542 from day 2. Samples were harvested on day 3 for ChIP.

2025-02-13 | E-MTAB-14280 | biostudies-arrayexpress

Functional CRISPR Screen Identifies AP1-associated Enhancer regulating FOXF1 to modulate Oncogene-Induced Senescence

Project description:We carried out CRISPR-Cas9 functional screen focused on AP-1 bound enhancers that are activated upon oncogenic stress. The screen discovered Enh.AP1.OIS1 - an enhancer bound by AP1 that is required for activation of oncogene-induced senescence (OIS) response. We applied RNA-seq analysis to RASG12V-activated BJ cells transduced with either sgRNAs targeting AP-1 bound enhancer (sgRNA-69 or sgRNA71), or sgRNA targeting the target gene FOXF1 or non-targeting control sgRNA (sgNT)

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Expression dynamics of HAND1/2 in in vitro human cardiomyocyte differentiation

Project description:Hand1 and Hand2 are transcriptional factors, and knockout mice of these genes show left and right ventricular hypoplasia, respectively. However, their function and expression in human cardiogenesis are not well studied. To delineate their expressions and assess their functions in human cardiomyocytes (CMs) in vitro, we established two triple reporter human induced pluripotent stem cell lines, HAND1mCherry, HAND2EGFP, with MYH6-driven iRFP670 or constitutive tagBFP expression and investigated their expression dynamics during cardiac differentiation. On day 5 of the differentiation, HAND1 expression marked cardiac progenitor cells. We profiled the CM subpopulations on day 20 with RNA-sequencing and found that mCherry+ CMs showed higher proliferative ability than mCherry- CMs and identified CD105 as a surface marker of highly proliferative CMs. Finally, we revealed that LEF1 is a key regulator of proliferation and is repressively regulated by HAND1 and HAND2.

2021-07-22 | GSE156394 | GEO

Single cell RNA-seq to resolve heterogeneity in a cardiac differentiation timecourse from wild-type and HAND1-null hESCs

Project description:The aim was to assess the role of HAND1 in cardiac differentiation. The differentiation protocol was designed to promote cell diversity. For wild-type and HAND1-null cell lines we combined cells differentiating under different conditions: 50% control, 25% SB-treated (day 2-3) and 25% DMH1-treated (day 2-3). The addition of SB at day 2, inhibited SMAD2/3 phosphorylation and created a high BMP signalling bias to restrict cardiac differentiation in favour of other mesodermal lineages, whereas the addition of DMH1 at day 2, inhibited SMAD1/5/8 phosphorylation to create a high Activin signaling bias to promote the co-differentiation of endoderm. Samples were collected at days 3, 4, 5, 6, 7, 8 and 10 for wild-type and HAND1-null populations. An additional day 7 and 8 sample was included for the wild-type, which was generated in the same condition.

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ChIP-seq analysis of Hand1 and Foxf1 at different stages of directed cardiac differentiation of mouse embryonic stem cells

Project description:ChIP-seq analysis of Hand1 and Foxf1 at different stages of directed cardiac differentiation of mouse embryonic stem cells

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HAND1 Ubiquitination and Phosphorylation sites

Project description:pcDNA3-HAND1-his was transfected into HEK293T cells and harvested after 48 hours. IMAC was carried out under denaturing conditions to enrich for HAND1,and samples were loaded onto SDS-PAGE gel with HAND1 gel band excised and submitted to LC-MS/MS.

2020-12-18 | PXD022407 | Pride

E11.5 Hand1 myocardial knockout RNA-Seq

Project description:Aims: To examine the role of the basic Helix-loop-Helix (bHLH) transcription factor HAND1 in embryonic and adult myocardium. Methods and Results: Hand1 is expressed within the cardiomyocytes of the left ventricle (LV) and myocardial cuff between embryonic days (E) 9.5-13.5. Hand gene dosage plays an important role in ventricular morphology and the contribution of Hand1 to congenital heart defects requires further interrogation. Conditional ablation of Hand1 was carried out using either Nkx2.5 knockin Cre (Nkx2.5Cre) or a-myosin heavy chain Cre (aMhc-Cre) driver. Interrogation of transcriptome data via Ingenuity Pathway Analysis (IPA) reveals several gene regulatory pathways disrupted including translation and cardiac hypertrophy-related pathways. Embryo and adult hearts were subjected to histological, functional and molecular analyses. Myocardial deletion of Hand1 results in morphological defects that include cardiac conduction system defects, survivable interventricular septal defects (VSDs), and abnormal LV papillary muscles (PM). Resulting Hand1 conditional mutants are born at Mendelian frequencies; but the morphological alterations acquired during cardiac development result in, the mice developing diastolic heart failure. Conclusions: Collectively, these data reveal that Hand1 contributes to the morphogenic patterning and maturation of cardiomyocytes during embryogenesis and although survivable, indicate a role for Hand1 conduction system and papillary morphogenesis.

2019-06-01 | GSE128571 | GEO

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