RNA-seq of endothelial cells in wild-type and Shb iECKO mice

E0771.lmb tumors were grown after tamoxifen injection as previously described in wild-type and Shb iECKO (Shb flox/flox; Cdh5-CreERt2) mice, alternatively Shb ICKO (Shb flox/flox; Vav1-cre) mice. Mice were sacrificed on day 14-16 after tumor cell injection. The tumors were digested, CD45+-cells isolated by MACS microbeads (Miltenyi, Bergich Gladbach, Germany) and subsequently CD31+-cells isolated by fluorescence-activated cell sorting (FACS) after staining for CD31. RNA from EC CD31+-cells were subjected to bulk RNAseq. Overall design: Bulk EC RNAseq was performed on 11 wild type (from nine tumors) and 10 Shb iECKO (from eight tumors) samples in four separate experiments.