Figure 2: p53 is not required for 53BP1-mediated mitochondrial function.
From: 53BP1 contributes to regulation of autophagic clearance of mitochondria
(A) A diagram of the strategy for knocking out 53BP1 using the transcription activator-like effector (TALE) endonuclease. Exon 4 of 53BP1 was targeted in the region shown, and the two TALE nuclease binding sequences are underlined. (B) Sequencing of the 53BP1 gene from the selected 53BP1 knockout H1299 clone. Dashes indicate deleted bases. The predicted amino acid sequence is indicated based on the recommended description (see Materials and Methods). (C) Immunoblot analysis of wild-type cells (WT) and 53BP1 deletion mutant clone (KO). β-actin expression was used as an internal control. (D) 53BP1 WT and KO H1299 clones were stained with MitoTracker RedCMXRos and fluorescence images were obtained using confocal microscopy. Nuclei were visualized by DAPI staining. (E) 53BP1 WT and KO H1299 clones were incubated with MitoTracker Red CMXRos and mitochondrial mass determination was performed as described in Fig. 1C. Results are shown as mean ± SD (n = 3), **P < 0.01. (F) 53BP1 WT and KO H1299 clones were stained with TMRM for 15 min to determine ΔΨm (Mitochondrial membrane potential) as described in Fig. 1D. Results are shown as mean ± SD (n = 3), **P < 0.01.
