Osteoblasts isolated from sp7-GFP transgenics by FACS sorting of 4dpf zebrafish larvae

The transgenic zebrafish line Tg(sp7:sp7-GFP) (ulg071 Tg) was used to obtain fluorescent cells through FACS sorting. Two populations were identifed: P1 displaying low fluorescence, P2 displaying high fluorescence. Cells were collected and sumitted to RNA-Seq Overall design: A Tg(sp7:sp7-GFP) heterozygous transgenic parent was outcrossed with a wt parent to obtain a clutch of transgenic and non-transgenic offspring. Two populations with low (P1) and high (P2) fluorescence were separated. The collected cells from populations P1 and P2 (about 30-50000 cells) were immediately lysed in 0.5% Triton X-100 containing 2U/µl RNAsin and stored at -80°C. cDNAs were prepared from these lysed cells according to the SMART-Seq 2.0 protocol (Illumina, San Diego, CA, USA) for low input RNA sequencing, while libraries were prepared using the Nextera XT DNA library kit. Third-party re-analysis: Table_P2_Sanger_P1_Sanger_P1_P2.csv (our DESeq2 analysis of P1 and P2 4dpf larvae populations with Sanger dataset from Bioproject PRJEB7244 : ID;gene;baseMean P2_Sanger;log2FoldChange P2_Sanger;padj P2_Sanger;Sanger_baseMean;P2_baseMean;ID;gene;baseMean P1_Sanger;log2FoldChange P1_Sanger;padj P1_Sanger;Sanger_baseMean;P1_baseMean;ID;gene;baseMean;log2FoldChange P1_P2;padj P1_P2;P2_baseMean;P1_baseMean). Table_Counts_P1_P2_Sanger.csv (the counts table complete with the Sanger data).