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. 2011 Mar;4(2):228–239. doi: 10.1242/dmm.006130

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© 2011. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms

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Fig. 7. — Blocked M-CSF signal transduction in induced SHP-2-deficient mice. (A) Bone marrow cells from a ptpn11^fl/fl ert2-cre mouse and a littermate ptpn11^fl/fl mouse (both treated with tamoxifen 10 days beforehand at 7 weeks of age) were cultured in wells of a 24-well plate (1×10⁶ cells/well) in the presence of M-CSF. After 5 days, macrophages in wells were harvested and counted. Depicted is the mean number of macrophages ± 1 s.e.m. (n=4). Results are representative of four repeat experiments. Statistical significance was determined by two sample Student’s t-test. **P<0.005. (B) Western blots showing expression of SHP-2 in bone marrow cells (freshly isolated or after culture in M-CSF for 5 days) from a ptpn11^fl/fl ert2-cre mouse and a littermate ptpn11^fl/fl mouse (both treated with tamoxifen 3 weeks beforehand at 6 weeks of age). NS, non-specific band. Blots were stripped and reprobed with an anti-GAPDH antibody to verify equivalent protein loading. Similar results were obtained in five independent experiments. (C) Lineage-negative bone marrow cells from a ptpn11^fl/fl ert2-cre mouse and a littermate ptpn11^fl/fl mouse (both treated with tamoxifen 10 days beforehand at 6 weeks of age) were stimulated with M-CSF for the indicated times (in minutes). Activation of AKT was determined by western blotting of whole-cell lysates using a phospho-specific anti-AKT antibody. Blots were reprobed with an anti-AKT antibody to verify equal loading. Similar results were obtained in three repeat experiments.