RNA seq from peritoneum, peripheral blood, spleen and tumors from TC-1 grafted mice

Splenocytes, peripheral blood cells, peritoneal cells and tumor cells were washed with PBS and were counted. Cells were centrifuged at 400 g for 5 min and supernatant was removed. Cells pellet (< 3 x 106 cells) was resuspend in 350 µl of RLT buffer (Qiagen). RNA was extracted with RNeasy Mini kit (Qiagen) according to manufacturer's protocol. mRNA library preparation was realized following manufacturer's recommendations (KAPA mRNA HyperPrep ROCHE). Library purity/integrity were assessed using an Agilent 2200 Tapestation (Agilent Technologies, Waldbrunn, Germany). Final 7 samples pooled library prep were sequenced on Nextseq 500 ILLUMINA with MidOutPut cartridge (2x130Millions of 75 bases reads), corresponding to 2x18Millions of reads per sample after demultiplexing.