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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Mol Cancer Ther. 2013 Jul 3;12(9):1738–1748. doi: 10.1158/1535-7163.MCT-13-0049-T

Figure 2. #220–324 inhibited SHP2-mediated cell signaling and cellular proliferation.

Figure 2

(A) Ba/F3 cells were cultured in IL-3 (1.0 ng/mL) containing medium supplemented with #220–324 at the indicated concentrations or control DMSO. Cell numbers were determined 72 hrs later using the One Solution Cell Proliferation Assay kit. Experiments were performed three times with similar results obtained. Data shown are mean±S.E.M. from one experiment. (B) The cell cycle of Ba/F3 cells was analyzed 24 hrs after the treatment of #220–324 at the indicated concentrations. Percentages of the cells at G1, S, and G2/M phases were quantified by flow cytometric analyses. Experiments were repeated three times. Results shown are mean±S.E.M. from one experiment. (C) Ba/F3 cells were deprived of IL-3 overnight. Cells were treated with #220–324 (30 μM) for 3 hrs and then stimulated with IL-3 (2.0 ng/mL) for the indicated times. Whole cell lysates were prepared. The levels of p-Erk, p-Akt, p-Stat5, p-Jak2 were determined by immunoblotting analysis. Blots were striped and reprobed with anti-Erk, anti-Akt, and anti-Jak2, and anti-Stat5 antibodies to check for protein loading. Experiments were repeated three times. Representative results from one experiment are shown.