Apoptotic morphology and DNA fragmentation of ECFCs incubated with nRCAS1.

Day 7 ECFCs were incubated for 24 hours at 37°C in liquid media with 15% FCS, 15% human AB serum, rhSCF (20 ng/mL) and rhEPO (2 U/mL) with, or without nRCAS1. Then, cytospin specimens of the cells were prepared and stained with May-Giemsa. (A) Immature erythroid cells after liquid culture without nRCAS1. (B) Apoptotic cells after liquid culture with nRCAS1. (C) DNA samples were isolated from day 7 ECFCs incubated with, or without nRCAS1 for 24 hours. DNA fragmentation of ECFCs exposed to RCAS1 was shown by neutral pH and 1.6% agarose gel electrophoresis, and stained with 0.5% ethidium bromide. Lane 1 displays size markers; lane 2 displays control cultures with rhEPO (2 U/mL), but without nRCAS1; lane 3 displays cultures with rhEPO plus nRCAS1.