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. 2013 Apr 4;2013:170261. doi: 10.1155/2013/170261

Figure 6.

Figure 6

P53-dependent pathway mediates the downregulation of NANOG by ugonin J treatment in MCF-7 cells. (a) MCF-7 cells were treated with 10 μM ugonin J for 6, 24, and 72 hours before protein extraction. Western blot probed for anti-ABCG2, NANOG, p53, phospho-p53 Ser15 and 392, and cleaved PARP and caspase 9 antibodies. Equal amounts of protein were used (40 µg per lane). (b) Relative percentage of NANOG+ population in MCF-7, p53-overexpressing MCF-7, and pifithrin-α (p53 inhibitor)-treated MCF-7 (0.9 × 105 cells in 12-well plates) were treated with 10 μM ugonin J and counted after 48 hours of culture (n = 3). **P < 0.01 versus J-treated control. (c) Pifithrin-α treatment rescued the reductive effect of Nanog. MCF-7 cells were treated with ALA and ugonin J for 72 hours. Nanog (red) and phospho-p53ser15 (blue) expression was analyzed by immunofluorescent staining.