Figure 6. #220–324 abrogated the growth of JMML myeloid progenitors with PTPN11^E76K/+ mutation.

(A) Splenocytes (2×10⁴ cells/mL) from a JMML patient with the PTPN11^E76K/+ mutation were plated in methylcellulose medium containing GM-CSF (1.0 ng/mL) and #220–324 at the indicated concentrations or control DMSO. Colonies were enumerated 14 days later and normalized against the number of colonies derived from the cells without #220–324 treatment. Three patient samples were tested in three independent experiments. Similar results were obtained in each. Data are presented as mean±S.E.M. from one patient sample. (B) Splenocytes from a JMML patient with the PTPN11^E76K/+ mutation were cultured in RPMI 1640 medium containing GM-CSF (1.0 ng/mL) and #220–324 at the indicated concentrations or control DMSO. Total cell numbers were determined and normalized against the number of the cells in the untreated group. Three patient samples were tested in three independent experiments. Similar results were obtained in each. Data are presented as mean±S.E.M. from one patient sample. (C) Apheresis peripheral blood cells (2×10⁴ cells/mL) from normal donors were plated in methylcellulose medium containing GM-CSF (1.0 ng/mL) and #220–324 at the indicated concentrations. Colonies were enumerated 14 days later and normalized against the number of colonies derived from the cells treated with control DMSO. Three samples were tested in three independent experiments. Similar results were obtained in each. Data are presented as mean±S.E.M. from one sample.