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Figure 3 | Molecular Cancer

Figure 3

From: ISG20L1 is a p53 family target gene that modulates genotoxic stress-induced autophagy

Figure 3

Knockdown of ISG20L1 does not affect 5-FU-induced apoptotic cell death. (a) RKO colon cancer cells were reverse transfected with control (siCon) or ISG20L1 targeting siRNA. Three days after transfection the cells were treated with 5-FU for 48 h and harvested for Western analysis. The Western blot (left and middle panel) is representative of three independent experiments and shows no change in caspase-3 or PARP cleavage after knockdown of ISG20L1 at either baseline 0 h, where there is low level, basal ISG20L1 expression or after 5-FU treatment. The right panel shows DNA laddering as observed on an ethidium bromide-stained gel, a characteristic marker of apoptosis, from the same samples as above treated with 5-FU. No difference in DNA laddering was evident (lanes are loaded evenly for DNA content). Positive control (+) was obtained from the Roche DNA laddering kit and represent U937 cells treated with camptothecin. (b) Flow cytometric analysis was performed on the samples described above at 48 h after 5-FU treatment and sub-G1 percentage was calculated.

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