Extended Data Figure 6. Cytokines/chemokines are aberrantly produced by Ptpn11^E76K/+ MSPCs.

a, Spleen tissues freshly dissected from Ptpn11^E76K/+Mx1-Cre⁺ and Ptpn11^+/+Mx1-Cre⁺ mice (n = 3 mice per group) 12 weeks after pI–pC administration were gently smashed in PBS (0.1 g tissue per 1.0 ml). Supernatant collected was processed for cytokine–chemokine array analyses with the Mouse Cytokine Antibody Array Kit following the instructions provided by the manufacturer. Representative results from one pair of the mice are shown. b, MSPCs (CD45⁻Ter-119⁻CD31⁻CD140α⁺Sca-1⁺) were freshly isolated from paired Ptpn11^E76K/+Nestin-Cre⁺ and Ptpn11^+/+Nestin-Cre⁺ mice at 7–8 months old by FACS. Total RNA was extracted and processed for RNA-sequencing analyses as described in Methods. The correlation coefficient between the two groups was 0.954, verifying that the method was accurate (left). Genes with more than 2.0 fold increased (in red) or decreased (in green) mRNA levels are shown on the right. Secreted protein factors are indicated. c, HSCs sorted from wild-type C57BL/6 mice were cultured in the presence of IL-1β (10 ng ml⁻¹), CCL3 (20 ng ml⁻¹), CCL4 (20 ng ml⁻¹), or CCL12 (20 ng ml⁻¹). Six days later, cells were collected and analysed for Mac-1⁺ myeloid cells, F4/80⁺ macrophages, and CD115⁺ monocytes by FACS. Data presented are mean ± s.d. of four independent experiments; ***P < 0.001. Source Data for this figure are available online.