Figure 4. Mutant MEFs and myeloid progenitors with the PTPN11^E76K/+ mutation were more sensitive to #220–324 than WT cells.

(A) WT and PTPN11^E76K/+ MEFs were cultured in 10% FBS-containing DMEM supplemented with #220–324 at the indicated concentrations. DMSO-treated cells were included as negative controls. Cell numbers were determined using the One Solution Cell Proliferation Assay kit 72 hrs later. Experiments were repeated three times. Similar results were obtained in each. Data shown are mean±S.E.M. from one experiment. (B) Bone marrow cells (2×10⁴ cells/mL) harvested from PTPN11^E76K/+/Mx1-Cre⁺ and PTPN11^+/+/Mx1-Cre⁺ mice 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 7 days later and normalized against the number of colonies derived from WT control cells without #220–324 treatment. Representative results from two independent experiments are shown. Data are presented as mean±S.E.M. (C) Bone marrow cells (5×10⁴ cells/mL) harvested from PTPN11^E76K/+/Mx1-Cre⁺ mice were plated in methylcellulose medium (without GM-CSF) supplemented with #220–324 at the indicated concentrations or control DMSO. Colonies derived in the cytokine-free medium were enumerated 7 days later and normalized against the number of colonies derived from the cells without #220–324 treatment. Representative results from two independent experiments are shown. Data are presented as mean±S.E.M.