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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
53148 SRR9692022 SRX6451056 SRS5104327 SRP214773 PRJNA554788 Gene expression analysis in the zebrafish fractured scale GSE134330 Transcriptome Analysis Bone is a connective tissue which undergoes continuous remodeling including bone resorption and formation by osteoclasts OCs and osteoblasts OBs respectively. Recently extracellular vesicles EVs are increasingly appreciated as a regulator of intercellular communication between OCs and OBs. The zebrafish scale is a thin membranous bone embedded in the skin and consists of OBs OCs and bone matrix providing an elegant model to visualize OC OB communication. Here we developed a double transgenic zebrafish line trap:GFP; osterix:mCherry which expresses GFP and mCherry under the control of the OC specific trap tartrate resistant acid phosphatase and OB specific osterix enhancer respectively. Utilizing this double transgenic line in combination with Hoechst 33342 Hoe staining we isolated four distinct fractions from the scale at 1 day post fracture by flow cytometry GFP– mCh+ Hoehigh OB fraction GFPlow mCh+ Hoehigh OC precursor pOC fraction GFPhigh Hoehigh mature OC mOC fraction and mCh+ Hoe– OB derived extracellular vesicle EV fraction. In this study we have performed RNA seq analysis using these four fractions to examine the gene expression profile of each fraction. Overall design: mRNA profiles of isolated OBs pOCs mOCs and OB derived EVs in the zebrafish fractured scales were generated by Quartz seq using Illumina NextSeq500.   pubmed:32327701   EV 2 GSM3942348   source name:osteoblast derived extracellular vesicles|age:adult|genotype:wild type|tissue:Sorted from the fractured sclae|cell type:osteoblast derived extracellular vesicles|molecule type:Amplified RNA EV 2 Illumina RTA software was used for base calling. Sequence reads were mapped to the zebrafish reference genome GRCz11 using HiSAT2 version 2.1.0. Reads per million RPM were calculated using the Subread ver. 1.6.4. Genome build: GRCz11 Supplementary files format and content: Reads per million RPM values for each sample. osteoblast derived extracellular vesicles   Total RNAs were extracted from sorted cells using RNeasy Mini Kit QIAGEN. Cells were directly sorted in a lysis buffer containing 1 μg/mL of polyinosinic polycytidylic acid and total RNA was extracted using RNeasy Mini Kit. Reverse transcription RT was performed using Super Script III Thermo Fisher Scientific and an RT primer which contains oligo dT T7 promoter and PCR target region sequences. post digestion of remaining RT primers by exonuclease I Takara a poly A tail was added to the 3’ ends of the first strand cDNAs using terminal transferase Sigma. The second strand DNA was then synthesized using MightyAmp DNA polymerase Thermo Fisher Scientific and a tagging primer which contains oligo dT and PCR target region sequences. PCR amplification was performed using a suppression primer which allow to amplify small size DNA that contains complementary sequences at both ends of the template DNA. The amplified double strand cDNA was purified using QIAquick PCR Purification Kit Qiagen. Library preparation was performed using Nextera XT DNA Library Preparation Kit illumina.   age:adult|genotype:wild type|tissue:Sorted from the fractured sclae|cell type:osteoblast derived extracellular vesicles|molecule type:Amplified RNA GSM3942348 GSM3942348: EV 2; Danio rerio; RNA Seq GSM3942348   1 Total RNAs were extracted from sorted cells using RNeasy Mini Kit QIAGEN. Cells were directly sorted in a lysis buffer containing 1 μg/mL of polyinosinic polycytidylic acid and total RNA was extracted using RNeasy Mini Kit. Reverse transcription RT was performed using Super Script III Thermo Fisher Scientific and an RT primer which contains oligo dT T7 promoter and PCR target region sequences. post digestion of remaining RT primers by exonuclease I Takara a poly A tail was added to the three prime ends of the first strand cDNAs using terminal transferase Sigma. The second strand DNA was then synthesized using MightyAmp DNA polymerase Thermo Fisher Scientific and a tagging primer which contains oligo dT and PCR target region sequences. PCR amplification was performed using a suppression primer which allow to amplify small size DNA that contains complementary sequences at both ends of the template DNA. The amplified double strand cDNA was purified using QIAquick PCR Purification Kit Qiagen. Library preparation was performed using Nextera XT DNA Library Preparation Kit illumina.   RNA-Seq TRANSCRIPTOMIC cDNA PAIRED ILLUMINA NextSeq 500   SRP214773     EV_2_R1.fastq.gz EV_2_R2.fastq.gz fastq fastq 6759957000.0 22533190.0 GSM3942348 r1 0:150 1:150 A:1706593126;C:1699056754;G:1646763192;T:1707067337;N:476591 150 150     1706593126 1699056754 1646763192 1707067337 476591 SRX6451056 SRS5104327 SRA921382 GEO Kanazawa University 2 0.94501 0.94549 0.02885 0.03033 0.80458 0.80957 0.48799 0.50474 150 150 B B biological fallback assumption illumina nextseq 3prime poly_a nextera sc single_cell_plate quartzseq   Japan 2019-07-16 Adult Adult Bone or Cartilage Skeletal Element