Figure 1: Loss of 53BP1 induces mitochondrial aggregation and increases mitochondrial mass.
From: 53BP1 contributes to regulation of autophagic clearance of mitochondria
(A) Western blot analysis of 53BP1 from U2OS and MEF cells. Wild type (53BP1+/+) and 53BP1 knockout (53BP1−/−) MEF cells were prepared from embryos. U2OS cells were stably transfected with either non-targeted shRNA or two different 53BP1-targeted shRNAs. β-actin was included as an internal control. (B) Representative cells were stained with MitoTracker Red CMXRos to visualize mitochondria, and fluorescence images were obtained using confocal microscopy. Nuclei were visualized by DAPI staining. (C) Cells were incubated with MitoTracker Red CMXRos and the fluorescence was analyzed using flow cytometry. The fluorescence intensity was quantified and the percent change in mitochondrial mass was determined relative to control cells. Histograms from a representative replicate experiment are shown in the right panels. Results are shown as mean ± SD (n = 3), **P < 0.01. (D) Cells were incubated with TMRM (Tetramethyl Rhodamine Methyl Ester) for 15 min and intracellular fluorescence intensity was measured using flow cytometry. Fluorescence intensity was then quantified and the percent change in ΔΨm was determined. Results are shown as mean ± SD (n = 3), **P < 0.01.
