title: GSM4725123: SPF_0h_12; Mus musculus; RNA-Seq
library_construction_protocol: Mice were anesthetized. A 22-gauge catheter was inserted into the peritoneal cavity and the peritoneal cavity was flushed three times with 5mL warm PBS containing 2mM EDTA. The wash buffer was aspirated completely for each wash and discarded. Then, 5ml of digestion buffer (0.5% Trypsin, 24mM EDTA, Sigma) were injected and incubated for 10 Minutes while the mouse was kept warm under an infrared light. The digestion buffer was aspirated and the abdominal cavity was flushed three times with 5mL ice cold PBS containing 2mM EDTA and 3 % FCS to collect mesothelial cells. Cells were purified as previously described (22). In brief, cells were washed two times and then incubated with anti-mouse GPM6A (Clone Nr. 321, MBL, #D055-3) for 30 Minutes at 4°C. After centrifugation the cells were incubated with anti-rat IgG MicroBeads (Miltenyi, #130-048-502) and were purified by MACS Separation columns LS (Milteny, #130-042-401) according to their instructions.