Impaired Autophagy in Adult Bone Marrow CD34+ Cells of Patients with Aplastic Anemia: Possible Pathogenic Significance
Fig 6
Autophagy effects on the survival of CD34+ cells.
(A) and (B): CD34+ cells were treated with 3-MA or CQ, and the number of cells was counted at days 4, 7 and 10. 3-MA and CQ were able to inhibit proliferation of CD34+ cells in a concentration-dependent manner. The results shown are the mean±SEM of three independent experiments. (C) and (D): CFC assay. CD34+ cells were treated with 3-MA or CQ, and colonies were counted at day 10 and normalized to control conditions. 3-MA and CQ inhibited differentiation of CD34+ cells in a concentration-dependent manner. The results shown are the mean±SEM of three independent experiments. (E) and (F): Trypan blue assays revealing the percentage of dead cells. CD34+ cells were treated with 3-MA or CQ for 48 h prior to application of trypan blue. 3-MA and CQ induced the death of CD34+ cells in a concentration-dependent manner. The data represent the mean±SEM of three independent experiments. (G-I): Apoptosis detection by AnnexinV-FITC/PI assays. CD34+ cells were treated with 3-MA or CQ for 24 h, followed by staining with Annexin-V/PI. 3-MA and CQ induced the apoptosis of CD34+ cells in a concentration-dependent manner. G and H represent the mean±SEM of three independent experiments, with the Q2 quadrant (Annexin V+/PI+) and Q3 quadrant (Annexin V+/PI-) in I representing the percentages of early and late apoptotic cells, respectively. * P<0.05, ** P<0.01, *** P<0.001.