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. Author manuscript; available in PMC: 2021 Jul 12.
Published in final edited form as: Mol Pharmacol. 2021 May 3;100(1):57–62. doi: 10.1124/molpharm.121.000262

Figure 2. siRNA knockdown of VCP/p97 and the effect on TOP2-DNA complex processing to DSBs.

Figure 2

(A) VCP/p97 siRNA knockdown from triplicate experiments was tested by western blot. Each replicate represents the cells used in yH2AX and TARDIS experiments shown in Figures B and C. (B) Cells treated with VCP/p97 siRNA or control siRNA (CON) were exposed continuously to 10μM etoposide for up to 4 hours, and levels of protein-free DSBs were measured using the yH2AX assay. (C) VCP/p97 siRNA or control (CON) siRNA cells were treated with 100μM etoposide for 2 hours followed by 2 hours incubation in etoposide-free media. The TARDIS assay was used to measure levels of TOP2A- and TOP2B- DNA complexes at 0, 0.5, 1 and 2 hours after etoposide removal. Statistical significance was determined by two-way ANOVA and Bonferroni post hoc test.