Fig. 4

STAT3 and STAT5 inhibitors suppressed cytokine-independent TF-1(R140Q) cell proliferation. A TF-1(R140Q) cells were treated with C188-9 or NSC74859 for 24 h and subjected to western blotting for detection of the indicated proteins. β-Actin was used as a loading control. B The ratios of the mean gray value of phospho-STAT3(Tyr⁷⁰⁵) to the mean gray value of STAT3, (C) the ratios of the mean gray value of phospho-STAT5(Tyr⁶⁹⁴) to the mean gray value of STAT5 were calculated and the ratios of the treated groups to the control group were analyzed. The results represent three independent experiments with similar results, and the error bars indicate standard deviations. ***P < 0.001 compared with the control (0 µM C188-9). ^###P < 0.001 compared with the control (0 µM NSC74859). TF-1(R140Q) cells were treated with C188-9 (D), NSC74859 (E) or STAT5-IN-1 (F) for 3 days in the absence of GM-CSF, and cell viability was evaluated using MTT assays. The inhibition rate of three independent experiments with similar results, and error bars mark indicate standard deviations. **P < 0.01, ***P < 0.001 compared with the vehicle control. G TF-1(R140Q) cells were induced with 50 ng/mL EPO to differentiate for 7 days in the presence of AGI-6780 (0.2, 1µM), C188-9 (2.5, 5, 7.5 µM) or NSC74859 (50, 100, 200 µM), respectively. Afterwards, the cells were collected, and color change was photographed. The dose of “0” and/or “-” group represents the vehicle control