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. 2017 May 26;45(13):7760–7773. doi: 10.1093/nar/gkx443

Figure 5.

Figure 5.

Mitochondrial-targeted bKu proteins do not induce mtDNA depletion in human MCF7 cells. (A) MCF7 cell lines were grown for 4 days prior to treatment with Dox (pretreatment) and then harvested. Total DNA was isolated from a portion of the cells, and the remaining cells were propagated in the presence or absence of Dox for 2 weeks. During the 2-week period, each cell line was passaged after reaching ∼90% confluency. At the end of 2 weeks, cells were harvested to obtain total DNA and total cell extract (TCE). (B) Western analysis of TCEs was performed and the membranes were probed with anti-Flag or anti-actin antibody. An immunoblot representative of two independent experiments is shown. TetOn: MCF7 Tet-On; TreTight: MCF7 TreTight; cMtKu: MCF7 cMtKu; cMmKu: MCF7 cMmKu. (C) From total cell DNA, the relative mtDNA copy number was determined for each cell line in each group. The relative mtDNA copy number of each cell line in the treatment groups (+ and −Dox) was normalized to the respective cell line in the pretreatment group. Data are represented graphically as mean ± SD from three independent experiments.