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. 2019 Sep 9;17(9):526. doi: 10.3390/md17090526

Figure 2.

Figure 2

Ech A enhances PB-CD34+ cell expansion by suppressing the activation of p38-MAPK and JNK. 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 protein was subjected to western blot analysis with the indicated antibodies. β-actin served as the loading control. (B, C) Cells were pretreated with SB203580 (10 μM) or SP600125 (10 μM) for 4 hr. (B) Total cell number was measured 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. (C) Cells were treated with 10 μM 2′7′-dichlorodihydro-fluorescein diacetate for 30 min. The values in the 2′7′-dichlorofluorescein histograms indicate MFI.