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. 2010 Jan 20;101(5):1177–1185. doi: 10.1111/j.1349-7006.2010.01503.x

Figure 1.

Figure 1

 Cisplatin‐resistant cells are more sensitive to mithramycin than their parental cells and show decreased expression of Foxo3a transcription factor. (a) T24, T24/DDP10, KK47, and KK47/DDP20 cells were seeded into 96‐well plates. The following day, various concentrations of mithramycin were applied. After incubation for 48 h, cell survival was analyzed by a cytotoxicity assay. Cell survival in the absence of mithramycin corresponds to 1. All values are representative of at least three independent experiments. Boxes, mean; bars, ±SD. (b) After extraction of total RNA from T24, T24/DDP10, KK47, and KK47/DDP20 cells and synthesis of cDNA, quantitative real‐time PCR was performed using the primers and probes for Foxo3a and GAPDH. The Foxo3a transcript level was corrected for the corresponding GAPDH transcript level. All values represent at least three independent experiments. The level of Foxo3a transcript from T24 cells was defined as 1. Boxes, mean; bars, ±SD. Whole‐cell extracts and nuclear extracts of T24, T24/DDP10, KK47, and KK47/DDP20 cells were subjected to SDS‐PAGE, and western blotting was performed using the indicated antibodies. Immunoblots for β‐actin and Lamin B1 are shown as loading controls.