American Society of Hematology

Fig. 3.

$Fig. 3. p27 message and protein levels during myelopoiesis. (A) Persistence of p27 message in differentiating CD34+cells. Aliquots of cells grown in the presence of SCF, G-CSF, and IL-3 for 7 or 10 days were harvested for RNA and a Northern blot probed for p27 message as shown. (B) Freshly harvested UC CD34+cells were cultured in the presence of cytokines for 12 days. At days 0, 6, 9, and 12 aliquots of 1 million cells were directly lysed in laemmli buffer and fractionated on a 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel for Western blot analysis. Neutrophils (N) and lymphocytes (L) from the same cord sample were analyzed. p27 immunoblot is shown; p21 levels were undetectable in this assay. (C) p27 immunoblot of CD34+ blasts fractionated for nuclear (N) and cytoplasmic expression of p27.$

p27 message and protein levels during myelopoiesis. (A) Persistence of p27 message in differentiating CD34⁺cells. Aliquots of cells grown in the presence of SCF, G-CSF, and IL-3 for 7 or 10 days were harvested for RNA and a Northern blot probed for p27 message as shown. (B) Freshly harvested UC CD34⁺cells were cultured in the presence of cytokines for 12 days. At days 0, 6, 9, and 12 aliquots of 1 million cells were directly lysed in laemmli buffer and fractionated on a 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel for Western blot analysis. Neutrophils (N) and lymphocytes (L) from the same cord sample were analyzed. p27 immunoblot is shown; p21 levels were undetectable in this assay. (C) p27 immunoblot of CD34⁺ blasts fractionated for nuclear (N) and cytoplasmic expression of p27.

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