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. Author manuscript; available in PMC: 2020 Feb 14.

Published in final edited form as: J Med Chem. 2018 Dec 5;62(3):1125–1137. doi: 10.1021/acs.jmedchem.8b00513

Figure 4. — (A) Ba/F₃ cells were cultured in IL-3 (1.0 ng/mL) containing medium supplemented with 1 at the indicated concentrations or DMSO. Cell numbers were determined 48 hours later using a One Solution Cell Proliferation Assay kit. Experiments were performed three times with similar results obtained in each. Data shown are the mean ± SD of triplicates from one representative experiment. (B) Ba/F₃ cells were deprived of IL-3 overnight. Cells were treated with 1 (20 µM) for 4 hours and then stimulated with IL-3 (2.0 ng/mL) for the indicated times. (C) Ba/F₃ cells were deprived of IL-3 overnight. Cells were treated with 1 at the indicated concentrations for 4 hours and then stimulated with IL-3 (2.0 ng/mL) for 10 min. Whole cell lysates were prepared. Levels of p-Erk, p-Akt, p-Jak2, and p-Stat5 were determined by immunoblotting analyses. Blots were striped and reprobed with anti-Erk, anti-Akt, anti-Jak2, and anti-Stat5 antibodies to check for protein loading. Experiments were repeated independently three times. Representative results from one experiment are shown.

Figure 4. — (A) Ba/F₃ cells were cultured in IL-3 (1.0 ng/mL) containing medium supplemented with 1 at the indicated concentrations or DMSO. Cell numbers were determined 48 hours later using a One Solution Cell Proliferation Assay kit. Experiments were performed three times with similar results obtained in each. Data shown are the mean ± SD of triplicates from one representative experiment. (B) Ba/F₃ cells were deprived of IL-3 overnight. Cells were treated with 1 (20 µM) for 4 hours and then stimulated with IL-3 (2.0 ng/mL) for the indicated times. (C) Ba/F₃ cells were deprived of IL-3 overnight. Cells were treated with 1 at the indicated concentrations for 4 hours and then stimulated with IL-3 (2.0 ng/mL) for 10 min. Whole cell lysates were prepared. Levels of p-Erk, p-Akt, p-Jak2, and p-Stat5 were determined by immunoblotting analyses. Blots were striped and reprobed with anti-Erk, anti-Akt, anti-Jak2, and anti-Stat5 antibodies to check for protein loading. Experiments were repeated independently three times. Representative results from one experiment are shown.