Figure 2.

Generation of a genetically-engineered mouse model for global, inducible deletion of Ptpn6. (A) Shp1 protein relative to total Erk2 protein in peripheral blood cells analyzed over time, from Ptpn6^fl/fl and Ptpn6^fl/fl ERT2-Cre mice that had been treated with tamoxifen (200 mg/kg bid for 4 days, represented by shaded area). Data is representative of at least three independent experiments with 4–5 mice pre group. (B) Representative immunoblot from A of Shp1 and Erk2 protein levels in whole cell lysates of peripheral blood cells from indicated mice on day 18 following tamoxifen treatment. Lysates were analyzed by SDS-PAGE followed by immunoblotting with indicated antibodies. (C) Flow cytometric analysis of Ptpn6 mRNA levels in the indicated subsets of immune cells from Ptpn6^fl/fl and Ptpn6^fl/fl ERT2-Cre mouse spleens on day 12 after MC38 tumor implant (day 19 after initial tamoxifen dose). Data is from one experiment with n = 3 mice/group. (D) Analysis of Shp1 protein levels by immunoblot in mouse spleen, bone marrow, and liver tissue lysates. Organs were isolated from MC38 tumor-bearing Ptpn6^fl/fl and Ptpn6^fl/fl ERT2-Cre mice at study endpoint and tissue lysates were homogenized, then analyzed by SDS-PAGE followed by immunoblotting with indicated antibodies. Quantitated Shp1 protein was normalized to total Erk protein. Data is representative of three independent experiments with 4–10 mice/group. Error bars represent SEM, ***p < 0.001, ****p < 0.0001.