title: GSM4219519: LysMCre-Cripto-deficient macrophages Rep 1; Mus musculus; RNA-Seq
library_construction_protocol: To induce muscle damage, cardiotoxin was injected in the tibialis anterior, gastrocnemius and quadriceps muscles for 5 days. Muscles were removed and separated from the bones, and dissociated by enzymatic digestion using 3U/mL Dispase II, 0.5U/mL Collagenase A, 10μg/mL DNAse I , 400μM CaCl2 and 5μM MgCl2 in PBS, for 75 minutes in a shaking bath at 37 °C. Cells were then washed with DMEM and filtered with 100 µm and 70 µm cell strainers. Following filtration, red blood cells were removed, and the remaining cells were re-suspended in PBS 2% FBS (FACS buffer). Macrophages were blocked using purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™; 1:100) for 15 minutes at 4 °C and then incubated with the indicated primary antibodies for 30 minutes at 4 °C. After washing with ice-cold PBS, cells were either analysed or sorted with a FACSAria II (BD Biosciences.  RNA was harvested using the combination of QIAzol Lysis Reagent and RNAeasy minikit columns (Qiagen) following the manufacturer's instructions. cDNA library preparation and amplification were performed from 200 ng total RNA using NEBNext Ultra RNA Library Prep Kit for Illumina.
attributes.tag: GEO Accession
attributes.value: GSM4219519