Pancreatic cancer organoid
Pancreatic cancer organoid
(Mouse organoids from Bag6 WT, Bag6 KO and PBS control groups were          collected in Eppendorf tubes (n=3 per each group). Organoids were          dissociated in cold PBS to remove the Matrigel. Cells were washed          twice with cold PBS and centrifuged at 300 xg for 5 minutes at 4°C.     Pancreatic cancer organoids were derived from the KPC mouse model   according to published procedures (1). The KPC organoids (5,000 cells   in 20 µL matrigel per 96 well) were cultured in organoid growth medium   (1) plus the secretome (ratio 1:1) derived from mouse mast cells   pre-treated according to the following conditions: 1. PBS: Mast cells pre-treated with PBS (control). 2. KO-EVs: Mast cells pre-treated with Bag6 KO EVs. 3. WT-EVs: Mast cells pre-treated with Bag6 WT EVs.  After 5 days, phase contrast images were taken by confocal microscope   (Leica DMI3000 B) and cell percentage was assessed using the Cell   Titer 3D GLO kit (Promega, Walldorf, Germany), control PBS was set to   100%. RNA was extracted at the same timepoint.  References:  (1). Boj SF, Hwang CI, Baker LA, et al. Organoid models of human and   mouse ductal pancreatic cancer. Cell 2015;160:324-38. Subsequently, the cells were lysed, and total RNA was isolated using          the RNeasy Plus kit (Qiagen, Hilden, Germany, [Cat. No. / ID:  74034])          according to the manufacturer's instructions).  Lexogen QuantSeq according to Manufacture's Instructions