run_metadata: 63227

This data as json

rowid	run.accession	experiment.accession	sample.accession	study.accession	bioproject	study.title	study.alias	study.type	study.abstract	study.attributes	study.PMIDs	sample.description	sample.title	sample.alias	sample.centername	sample.attributes	GEOsample.title	GEOsample.dataprocessing	GEOsample.source	GEOsample.treatmentprotocol	GEOsample.extractprotocol	GEOsample.growthprotocol	GEOsample.characteristics	GEOsample.accession	experiment.title	experiment.alias	experiment.library_name	experiment.design_description	experiment.library_construction_protocol	experiment.attributes	experiment.library_strategy	experiment.library_source	experiment.library_selection	experiment.library_layout	experiment.platform	experiment.instrument_model	experiment.spot_descriptor	experiment.study_ref	run.title	run.attributes	run.filename	run.semantic_name	run.total_bases	run.total_spots	run.alias	run.read_lengths	run.base_counts	run.r1_length	run.r2_length	run.r3_length	run.r4_length	run.Acount	run.Ccount	run.Gcount	run.Tcount	run.Ncount	run.experiment	run.pool_member	submission.accession	submission.srasource	submission.bioprojectsource	seqdetective.n_mates	seqdetective.mapping_rate.mate1	seqdetective.mapping_rate.mate2	seqdetective.nofeature_rate.mate1	seqdetective.nofeature_rate.mate2	seqdetective.sparsity.mate1	seqdetective.sparsity.mate2	seqdetective.pos_strand_rate.mate1	seqdetective.pos_strand_rate.mate2	seqdetective.readlen.mate1	seqdetective.readlen.mate2	seqdetective.judgement.mate1	seqdetective.judgement.mate2	seqdetective.judgement.reason	platform_family	instrument_generation	read_bias	selection_class	prep_kit	sc_or_bulk	tech_class	technology	tech_variant	submission.bioprojectsource.country	earliest_date	devstage_curation	devstage_curation_coarse	tissue_curation	tissue_curation_coarse
63227	SRR13660515	SRX10052200	SRS8215328	SRP305389	PRJNA700672	Endothelial progenitor migration and differentiation is regulated by distinct transcriptional effectors of Cloche/Npas4l	GSE166396	Transcriptome Analysis	Embryonic cells including endothelial progenitors undergo extensive migration and differentiation events; however whether and how these processes are interrelated remains unclear. The transcription factor Npas4l is necessary for endothelial specification in zebrafish by inducing the expression of the transcription factor genes etsrp tal1 and lmo2. We generated a knock in reporter in the npas4l locus to visualize endothelial progenitors and their derivatives in wild type and mutant embryos. We find that in npas4l mutants npas4l reporter expressing cells do not migrate to the midline and instead contribute to skeletal muscle and pronephric tubules. Investigating the Npas4l transcriptional effectors we find that npas4l reporter expressing cells in tal1 mutants fail to migrate while those in etsrp mutants migrate but fail to differentiate. In lmo2 mutants npas4l reporter expressing cells migrate and differentiate but many express pronephric tubule markers. Altogether these data reveal the complex regulation of endothelial progenitor migration differentiation and fate restriction. Overall design: Comparison of npas4l reporter positive cells from heterozygous and homozygous npas4l mutant embryos using single cell RNAseq on sorted cells.				npas4l het	GSM5070575		tissue:npas4l reporter positive cells\|cell type:sorted npas4l reporter positive cells\|Stage:20 hpf	npas4l het	Reads were aligned against the zebrafish genome DanRer11 and counted by StarSolo followed by secondary analysis in Annotated Data Format. Preprocessed counts were further analysed using Scanpy. Basic cell quality control was conducted by taking the number of detected genes and mitochondrial content into consideration. We removed only 16 cells in total that did not express between 1000 and 7000 genes or had an mitochondrial content less than 6%. Further we filtered genes if they were detected in less than 30 cells <0.3%. Raw counts per cell were normalised to the median count over all cells and transformed into log space to stabilise variance. We initially reduced dimensionality of the dataset using PCA retaining 50 principal components. Subsequent steps like low dimensional UMAP embedding and cell clustering via community detection were based on the initial PCA. Final data visualization was done by scVelo and cellxgene packages. Genome build: DanRer11 Supplementary files format and content: Mattonet adata obs.csv: basic anndata observations Sample n genes by counts log1p total counts pct counts is mito n genes detected leiden clustering Supplementary files format and content: Mattonet udata obs.csv: velocity anmdata observations initial size unspliced initial size spliced initial size n counts velocity self transition root cells end points velocity pseudotime	npas4l reporter positive cells		Tails from 150 npas4l heterozygous npas4lbns423/+ and trans heterozygous npas4lbns423/bns297 mutant embryos were cut at the anterior end of the yolk extension at 20 hpf in DMEM/F10+5%FBS+0.01% tricaine on agarose coated plates. The cells were dissociated using the Pierce Cardiomyocyte dissociation kit according to manufacturer instructions and FACS sorted for GFP fluorescence using an AriaIII sorter and DAPI as indicator for dead cells. The cell suspensions were counted with Moxi cell counter and diluted according to manufacturer’s protocol to obtain 5000 single cell data points per sample. Each sample was run separately on a lane in Chromium controller with Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 10xGenomics. Single cell RNAseq library preparation was done using standard protocol.		cell type:sorted npas4l reporter positive cells\|Stage:20 hpf	GSM5070575	GSM5070575: npas4l het; Danio rerio; RNA Seq	GSM5070575		1	Tails from 150 npas4l heterozygous npas4lbns423/+ and trans heterozygous npas4lbns423/bns297 mutant embryos were cut at the anterior end of the yolk extension at 20 hpf in DMEM/F10+5%FBS+0.01% tricaine on agarose coated plates. The cells were dissociated using the Pierce Cardiomyocyte dissociation kit according to manufacturer instructions and FACS sorted for GFP fluorescence using an AriaIII sorter and DAPI as indicator for dead cells. The cell suspensions were counted with Moxi cell counter and diluted according to manufacturer's protocol to obtain 5000 single cell data points per sample. Each sample was run separately on a lane in Chromium controller with Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 10xGenomics. Single cell RNAseq library preparation was done using standard protocol.	GEO Accession:GSM5070575	RNA-Seq	TRANSCRIPTOMIC	cDNA	PAIRED	ILLUMINA	NextSeq 500		SRP305389			Kenny_scRNA_Het_BC.fastq.gz Kenny_scRNA_Het_cDNA.fastq.gz	fastq fastq	21384880020.0	254581905.0	GSM5070575 r1	0:28 1:56	A:5828925615;C:4816199698;G:4885826464;T:5847660976;N:6267267	28	56			5828925615	4816199698	4885826464	5847660976	6267267	SRX10052200	SRS8215328	SRA1193720	GEO	MPI for heart and lung research	2	0.00213	0.9288	0.00071	0.11864	0.99559	0.8312	0.43162	0.47914	28	56	T	B	sc-like readlen	illumina	nextseq	unknown	cdna_unspecified	unknown	sc	single_cell_droplet	10x		Germany	2021-02-08	Segmentation	Embryo	Multi-tissue	Multi-system