Transcriptomic profiling of KPM cell lines through RNA-Seq

Conditional mutant KRAS-expressing (LSL.KRASG12D; K) and TRP53-deleted (Trp53f/f; P) mice were intercrossed in all possible combinations, followed by intrapleural adenovirus-Cre injection. KRAS G12D; TRP53 f/f mice (KP mice) developed multifocal, lethal Malignant Pleural Mesothelioma that invaded the lungs, the chest wall and the mediastinal structures with a median overall survival of 41 days. Multiple cell lines were derived from primary MPM from KP mice. This was done by isolating, in sterile conditions, cells from the tumors of the KP mice and culturing them in minimum supplemented medium (DMEM+10% FBS + 1% PEN/STREP) for over 6 months, for over 30 passages.The WT mesothelial cells were isolated through a lavage procedure at the pleural space. Wt C57BL/6 mice were sacrificed. In order to wash the pleural space 1ml of sterile saline was injected and immediately withdrawn through the diaphragm. Subsequently 1ml of trypsin 1% was injected to the pleural space. We incubated for 8 minutes and withdrew the trypsin which contained the mesothelial cells. The cells were pelleted by centrifugation at 500g for 5 minutes. Compare differential gene expression between naïve mouse mesothelial cells and mouse malignant pleural mesothelioma cells.Alignment of sequencing traces for visual exploration and visualizzation of trascripts alteration.