Figure 3.

Echinochrome A (Ech A) upregulates PB-CD34⁺ cell expansion by activating PI3K/Akt pathway. Cells were treated with 10 μM Ech A for 1 day (PBMCs) or 4 days (PB-CD34⁺ cells). For NAC treatment, cells were treated with 5 mM NAC for 4 h, washed, suspended in complete medium, and incubated for an additional 1 day (PBMCs) or 4 days (PB-CD34⁺ cells). (A) Total cell lysates for each condition were immunoblotted with the indicated antibodies. (B) Cell were pretreated with LY294002 (10 μM) for 4 h. Total cell number was determined using the ADAM-MC automated mammalian cell counter. For flow cytometric immunophenotypic analysis, cells were stained with CD34-PE, CD38-FITC, CD45-APC, and 7-AAD. Each value was expressed as the mean ± SEM of three independent experiments.