Figure 4. NAC treatment decreases cytokine signaling in Ptpn11 GOF mutant cells.

Four-week-old Ptpn11^+/+/Mx1-Cre+ and Ptpn11^{E76K neo/+}/Mx1-Cre+ mice were treated with pI-pC as described in Figure 1B. Twelve weeks after pI-pC treatment, bone marrow-derived macrophages were generated from bone marrow cells isolated from these animals. The macrophages were starved in serum and cytokine-free medium for 5 hours. NAC (1 mM) was added to the culture medium during the last two hours. These cells were then stimulated with GM-CSF (10 ng/ml) for the indicated periods of time. Whole cell lysates were prepared and examined for Erk and Akt activation by immunoblotting with anti-phospho-Erk and anti-phospho-Akt antibodies. Blots were stripped and reprobed with anti-Erk and anti-Akt antibodies to check for protein loading.