The effect of KitD814H on the proliferation and differentiation of ELM cells in response to SCF and/or Epo. (A) ELM-KitDH or ELM-Kit cells were cloned in methocel in the presence or absence of 10 ng/mL SCF and/or 5 U/mL Epo as described in Fig4B and the colonies were analyzed after 2 weeks. To analyze the numbers and sizes of colonies, the dishes were recorded using a scanner, and colonies were counted and sized using the NIH image program. The representative plotted data are shown as the average colony number from triplicate plates, with error bars showing maximum and minimum numbers. (B) To examine the extent of erythroid differentiation, the cells were incubated for 3 days in the presence or absence of 2 U/mL Epo or 10 ng/mL SCF, as indicated. Total RNA was then analyzed by Northern blotting using labeled cDNA probes encoding -globin or 7S ribosomal RNA.