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. 2012 Aug 10;7(8):e42921. doi: 10.1371/journal.pone.0042921

Figure 3. Doxorubicin treatment induces FILIP1L expression.

Figure 3

(A) U2OS cells were treated with 200 ng/ml doxorubicin and mRNA isolated 24 hours later for qPCR analysis. The twelve genes identified in the shRNA screen were tested for induction by doxorubicin. Expression of most genes was unaffected by doxorubicin treatment. However, two genes, expression of FILIP1L and HORMAD2 were significantly induced by doxorubicin treatment, particularly FILIP1L which showed >200-fold induction. (B) FILIP1L induction by doxorubicin impaired following ATM/ATR inhibition in U2OS. Doxorubicin treatment induces DNA damage that activates the ATM and ATR kinases. Caffeine (4 mM) was used to inhibit ATM and ATR. FILIP1L induction by doxorubicin is reduced by over 90% by treatment with caffeine. SAOS-2 cells, which unlike U2OS do not contain wild-type p 53, fail to induce FILIP1L following doxorubicin treatment.