Figure 5

siRNA northern blot analysis. (a) TS6-dTx5 ssiRNA was transfected into RKO cells. After 6 h, the cell culture medium was replaced. RNA was extracted using Trizol immediately and after an additional 24 and 48 h followed by northern blot analysis as described in the Methods section. (b) Thymidine rescue of ZD1694 toxicity. RKO cells were incubated with 3 nM ZD1694 in the presence or absence of 10 uM thymidine. Thymidine (10 μM) was also added 8 and 24 h after ZD1694 addition. After an additional 72 h, cell proliferation was quantified by the WST-1 assay. Cell growth values represent the mean ± standard deviation (SD) from three experiments performed in duplicate.