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Role of AMPK in human erythropoiesis
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Figure 5. GSK621-mediated AMPK activation specifically and drastically decreases the survival of mature GPAhigh erythroblasts. (A) GSK621-dependent AMPK acti- vation reduces the percentage of hemoglobinized cells. The percentage of hemoglobinized cells was determined by benzidine staining after incubation in the absence (vehicle) or presence of 20 mM GSK621 from day 0 to the indicated days. (B) GSK621-mediated AMPK activation blocked erythroblast differentiation. Erythroblasts were incubated in the absence (vehicle) or presence of 20 mM GSK621 from day 0 to the indicated days. Representative cytometry profiles for GPA/annexin V and band 3/a4 integrin are shown. For the GPA/annexin V profiles, annexin V-positive cells are shown in the upper part of the quadrant and GPAhigh cells in the right part. The percentage of GPAhigh cells and percentage of annexin V-positive cells are represented. Results are expressed as the mean ± SD of three independent experi- ments; ns: non-significant, *P<0.05. (C) Morphology was analyzed by May-Grünwald-Giemsa staining. Representative experiments at day 4, day 9 and day 14 are shown. d: day; GPA: glycophorin A.
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