title: GSM3477131: beclo treated_non amputated_rep2; Danio rerio; RNA-Seq
library_construction_protocol: Macrophages were sorted from Tg(mpeg1.4:mCherry-F) embryos as previously described (Rougeot et al., 2014; Zakrzewska et al., 2010). Dissociation was performed with 100-150 embryos for each sample at 4 hours post amputation (hpa) using Liberase TL (Roche) and stopped by adding Fetal Calf Serum (FCS) to a final concentration of 10%. Isolated cells were resuspended in Dulbeccoâ??s PBS (DPBS), and filtered through a 40 Î¼m cell strainer. Actinomycin D (Sigma Aldrich) was added  (final concentration of 1 Âµg/ml) to each step to inhibit transcription. Macrophages were sorted based on their red fluorescent signal using a FACSAria III cell sorter (BD Biosciences). The sorted cells were collected in QIAzol lysis reagent (Qiagen) for RNA isolation. Extracted total RNA was amplified using the SMART-seq V4 kit (Clontech) for sequencing. The RNA seq libraries generated with the SMART-seq V4 kit were sequenced using an Illumina HiSeq 2500 instrument according to the manufacturerâ??s instructions with a read length of 50 nucleotides
attributes.tag: GEO Accession
attributes.value: GSM3477131