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. 2021 Nov 2;11:21533. doi: 10.1038/s41598-021-00958-4

Figure 1.

Figure 1

ATP depletion induces translocation of EGFP-TOP2A and EGFP-TOP2B to the nucleoli. (a) Intracellular ATP levels after 2DG/CCCP treatment. HeLa cells were treated with 25 mM 2-deoxyglucose and 10 µM CCCP, and intracellular ATP levels were measured at the indicated time points. Average values with SD from 6 independent experiments were plotted. (b) Translocation of EGFP-TOP2A to the nucleoli by ATP depletion. Cells expressing EGFP-TOP2A were treated with 25 mM 2DG and 10 µM CCCP, and images were captured at the indicated time points. (c) Translocation of EGFP-TOP2B to the nucleoli by ATP depletion. An experiment was performed on EGFP-TOP2B as described in (b). (d, e) Nucleolus-to-nucleoplasm ratios of EGFP fluorescence. Cells expressing EGFP-TOP2A (d) or EGFP-TOP2B (e) were incubated in the presence and absence of 25 mM 2DG and 10 µM CCCP, and images were captured at 20 min. Green fluorescence in the nucleolus and nucleoplasm was quantified, and average values of the nucleolus-to-nucleoplasm ratio and SD were calculated (n = 30; *: statistically significant, d: p = 7.97E−14, e: p = 1.23E−14).