Mouse thrombopoietin tpo

Murine HSCs were collected and cultured following the methods described above in Cell culture section. Briefly, 1000/well LT-HSCs were sorted into 96-well U-bottomed plate, containing S-Clone SF-03 culture medium (EIDIA) supplemented with 10 ng/ml mouse stem cell factor (SCF) (R&D systems, Minneapolis, MN) and 100 ng/ml mouse thrombopoietin (TPO) (R&D systems) and 0.5% fatty acid -free bovine serum albumin (BSA) (Proliant Biologicals, Ankeny, IA). Subsequently, KIRA6 (Selleckchem; S8658), an ER stress inhibitor, were added into cultured LT-HSCs with final concentrations of 0.03 μM and 0.1 μM, respectively. After incubation at 37 °C for 20 h, cells were collected and stained with PI and Annexin V - APC probe (BD Pharmingen) following the protocol provided by the manufacturer. Apoptotic populations were detected and analyzed by flow cytometry Aria IIIu (BD Sciences).