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. 2010 Jul;12(7):527–538. doi: 10.1593/neo.92048

Figure 6.

Figure 6

ATF3 expression mediates, in part, the cytotoxic effects of cisplatin. (A) A549 cells treated with cisplatin (0–10 µg/ml) in the presence or absence of 50 µM SP600125, 25 µM UO126, and 10 µM SB203580 for 48 hours were assessed for cell cytotoxicity as measured by MTT activity. (B) A549 cells stably expressing shRNA against two separate ATF3 mRNA regions (shATF3-1 and shATF3-2) and GFP (negative control) were treated with cisplatin for 48 hours and were analyzed for MTT activity. For A and B, data are presented as a percentage of MTT activity where untreated cells were taken to be 100%. Error bars are representative of six independently treated samples. (C) A549 cells without treatment (untreated) or treated with 10 µg/ml cisplatin for 24 hours in the presence of MAPK inhibitors of 50 µM SP600125, 25 µM UO126, and 10 µM SB203580 and analyzed by Western blot analysis for PARP and actin as a loading control. (D) GFP-, shATF3-1-, and shATF3-2-expressing cell lines treated with or without 10 µg/ml cisplatin for 24 hours and analyzed by Western blot analysis for PARP, ATF3, and actin expression. (E) ATF3-/- and ATF3+/+ MEFs treated with cisplatin (0–8 µg/ml) and analyzed for MTT activity. Western blot analysis for ATF3 and actin in ATF3-/- and ATF3+/+ MEFs treated with 10 µg/ml of cisplatin for 24 hours (inset). Actin was used as a loading control. (F) ATF2-/- and ATF2+/+ MEFs treated with cisplatin (0–8 µg/ml) and analyzed for MTT activity.