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. 2022 Sep 23;9:1007064. doi: 10.3389/fmolb.2022.1007064

FIGURE 2.

FIGURE 2

Knockdown of NBS1 by siRNA increases the level of etoposide-induced TOP2-DNA complexes. Experimental conditions and methods were the same as Figure 1. (A) Western blot assessing NBS1 levels using antibody against NBS1. Semi-quantitative image analysis of immunoblots using ImageJ indicated that treatment with siNbs1 resulted in 85% knockdown of endogenous NBS1 (quantitation not shown). β-actin was used as a protein loading control. (B) ICE assay illustrating TOP2αcc in NBS1 knockdown cells. The panel illustrates a representative assay. (C) Densitometric analysis of all experiments comparing relative integrated densities of TOP2αcc signal amongst NBS1 knockdown and control cells treated with 2 μM, 10 and 50 μM etoposide. Integrated density of TOP2αcc signal of each group was normalized to that of control cells treated with 2 μM etoposide. * denotes p-value <0.05. Samples not marked with a bar did not result in a statistically significant difference. (D) ICE assay illustrating TOP2βcc. The panel illustrates a representative assay. (E) Densitometric analysis of all experiments comparing relative integrated densities of TOP2βcc signal amongst MRE11 knockdown and control cells.