Figure 2
Overexpression of P18 from D0 inhibits hematopoiesis in co-culture with AGM-S3 cells. (a) P18/hESCs were cultured with AGM-S3 cells, 1 × 104 CD34HighCD43− and CD34LowCD43− cells were sorted from non-induced co-cultures at D6, and then re–re-plated in FLHD medium in each well of a 48-well plate without DOX induction for 14 days. (b) P18/hESCs were treated with or without DOX from D0, D2, D4, or D6, and then subjected to flow cytometry at D8 using 7-AAD and anti-CD34/CD43 antibodies. The GFP+ fractions of non-induced co-cultures and of co-cultures treated with DOX were compared. Overexpression of P18 at the early stage, especially from D0, led to significantly reduced production of CD34highCD43−, CD34−CD43+, and CD34+CD43+ cells at D8. (c) When P18 was induced from D0 to D8 or only from D0 to D2, production of the CD34highCD43−, CD34−CD43+, and CD34+CD43+ populations were also reduced to a similar degree at D8. All results are expressed as means ± SD of the three repeated experiments, and p < 0.05 was considered significant (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).
