![Fig. 2. Effect of IFN-γ on apoptosis of ECFCs. / (A) Day 7 ECFCs were incubated in serum-free medium without rhEPO and rhIFN-γ (upper left), with rhEPO (10 U/mL) (upper right), with rhIFN-γ (1000 U/mL) (lower left), and both (lower right). After incubation for 16 hours, apoptosis was measured with PI and annexin V, using a flow cytometer. Data shows typical results of 12 replicates from 4 experiments. In each panel, the right lower quadrant (annexin V–positive and PI negative) indicates early apoptosis, and the right upper quadrant (annexin V– and PI positive) indicates late apoptosis. Both annexin V–positive fractions were assessed as apoptotic cells. (B) The day 7 ECFCs were labeled with [3H]thymidine and cultured in serum-free medium without rhIFN-γ (left), or with rhIFN-γ (1000 U/mL) (right), for 16 hours. Cellular DNA was isolated and analyzed by alkaline pH, 0.6% agarose gel electrophoreses. The sum of the radioactivity of fractionations 1-4 is designated as uncleaved DNA and is expressed as a percentage of the total radioactivity.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/95/12/10.1182_blood.v95.12.3742/8/bloo01209002x.jpeg?Expires=1722831479&Signature=DVpV45hoVkFZfPGybJ6ROuLTVLrMrRtdY9dblrr--Pk5B~K7NTdABQwzyCuImkHoix6sSRE63KYx9zPt7b3koqxyTQ9XGXJ0H1z~HMI8~5hmffddKtI4qZ9kK3reSbDxKVbvUgQ-4T8Sk~RlVn87ZUhl0zGj2UNdYkIWDHvBtwbf3KgX6eJlaH~8R30z6Sgr4ZhoFcQ1UHgokX3Og3~bnZBdkJVYxfgDT6hwgHFagWu-M47hwf-GMZSnQvjGCJijjcifPephPhCdn~1QQvRLtdHMTXxBj77G8hvI5gFPEex4vyRCHg23MyEAQTQzrkts3CVqt1nFtWB7I4ot~fMucg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Effect of IFN-γ on apoptosis of ECFCs.
(A) Day 7 ECFCs were incubated in serum-free medium without rhEPO and rhIFN-γ (upper left), with rhEPO (10 U/mL) (upper right), with rhIFN-γ (1000 U/mL) (lower left), and both (lower right). After incubation for 16 hours, apoptosis was measured with PI and annexin V, using a flow cytometer. Data shows typical results of 12 replicates from 4 experiments. In each panel, the right lower quadrant (annexin V–positive and PI negative) indicates early apoptosis, and the right upper quadrant (annexin V– and PI positive) indicates late apoptosis. Both annexin V–positive fractions were assessed as apoptotic cells. (B) The day 7 ECFCs were labeled with [3H]thymidine and cultured in serum-free medium without rhIFN-γ (left), or with rhIFN-γ (1000 U/mL) (right), for 16 hours. Cellular DNA was isolated and analyzed by alkaline pH, 0.6% agarose gel electrophoreses. The sum of the radioactivity of fractionations 1-4 is designated as uncleaved DNA and is expressed as a percentage of the total radioactivity.
