SRRM4 WT in HCT116, 24h induction
Cell extracts were harvested by direct addition of QIAzol lysis reagent (79306, Qiagen [WERFEN ESPAA, S.A.U. L'Hospitalet de Llobregat, Barcelona]) to the culture plate and incubation for 5 min at RT followed by collection in 1.5 mL eppendorf tubes. RNA was isolated using the miRNeasy mini kit (217004, Qiagen [WERFEN ESPAA, S.A.U. L'Hospitalet de Llobregat, Barcelona])) according to the manufacturers instructions. For isolation of RNA from mouse tumor tissue, tumors frozen in RNAlater stabilization solution (AM7020, ThermoFisher Scientific, Madrid, Spain) were thawed in QIAzol and disrupted using a Polytron PT1200E homogenizer (KINEMATICA INC, VWR), before continuing with the miRNeasy protocol. Purified RNA was stored at -80C before sequencing.