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Fig. 4 | BMC Cancer

Fig. 4

From: Inhibition of STAT3 enhances sensitivity to tamoxifen in tamoxifen-resistant breast cancer cells

Fig. 4

Inhibition of STAT3 shows a potent inhibitory effect on TamR cell survival. Cells were pre-incubated with 7 μM stattic for 4 h and then treated with 9 μM tamoxifen for 72 h. a Cell viability was measured using a CCK-8 kit. b For colony formation, cells were kept for 18 days and then stained with crystal violet and photographed. c After 4 h pre-incubation with the indicated concentration of stattic, cells were treated with tamoxifen for 24 h. Flow cytometric assay using annexin V-FITC and PI showed that stattic increased the total number of apoptotic cells. Histogram reported a summary of the apoptotic index. Annexin V (+) / PI (+) and annexin V (+) / PI (−): early and late apoptotic cells, annexin V (−) / PI (+): necrotic fraction, annexin V (−) / PI (−): live cells. d Lysates were immunoblotted for phospho-Src (p-Src), Src, phospho-EGFR (p-EGFR), EGFR, phospho-STAT3 (p-STAT3), STAT3, cyclin D1, cyclin D3 and cleaved PARP. The cropped blots are used in the figure. The membranes were cut prior to exposure so that only the portion of gel containing the desired bands would be visualized. GAPDH was used as the loading control. Data are expressed as mean ± SD and all experiments were performed in triplicate, independently. P values calculated using an unpaired t-test. *p < 0.05, **p < 0.01, ***p < 0.001

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