Figure 7.

Flowchart of the study design. EV depleted FBS was prepared by ultracentrifugation, ultrafiltration or purchased from commercial supplier. Detailed protocol for the preparation of EV depleted FBS is described in materials and methods section. EVs extracted by ultracentrifugation from 10 ml of FBS, EV depleted FBS and commercial defined media were characterized by NTA, TEM and WB. For small ncRNA sequencing, EVs and RNA were extracted using exoRNeasy Serum/Plasma Maxi Kit (Qiagen) from 3.5 ml of media containing 10% depleted FBS. Reads were mapped to both bovine and human genome and media derived small ncRNA were identified.