RNA-Seq of mus musculus: adult female Kupffer Cells
For sEVs-educating mice experiments, each mouse was injected with 20g of CRC-sEVs (SW620, LoVo) or normal colon cell-sEVs via tail vein every other day for 21 days. After 2 weeks, all mice were given gaseous anesthesia and were sacrificed and their livers excised and minced. The tissues were dispersed by 200U/ml type IV collagenase and vibrated in PBS at 37C for 30 min. Following digestion, each liver homogenate was filtered through a 70-m stainless steel wire mesh to remove undigested tissue, and the cell suspension was centrifuged at 300g for 5 min at 4C. The cells were harvested and washed, and the cell suspension was adjusted to 1 x 107 cells/mL in ice-cold PBS containing 10% FBS and monoclonal anti-F4/80 antibody [BM8] conjugated to FITC for 45 min (1:50, ab60343, Abcam). Flow cytometer (Influx, Becton Dickinson, USA) was used to detect two groups of F4/80-positive cells (FITC channel) with or without PKH26 (PE channel) positivity, respectively. The data were collected and analyzed flow cytometer (Aria/Influx/Canto 2, Becton Dickinson, USA).