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. Author manuscript; available in PMC: 2021 Jan 27.
Published in final edited form as: Cell Rep Phys Sci. 2020 Dec 9;1(12):100270. doi: 10.1016/j.xcrp.2020.100270

Figure 4. Perimitochondrial ENS for Mitochondrial Transgene Expression to Induce Mitophagy or Apoptosis.

Figure 4.

(A) Western blot analysis of FUNDC1 and LC3B levels in the mitochondria fraction and cytosolic fraction, respectively, of HeLa cells incubated with conditions of interest. VDAC1 serves as mitochondria protein loading control. FUNDC1 plasmid, 5 μg/mL; Mito-FLAG, 200 μM. Time: 2 days.

(B) Mito-FLAG perimitochondrial ENS delivers the FUNDC1 gene into mitochondria to induce mitochondrial morphology changes within 2 days.

(C) Detection of mitophagy via immunofluorescent staining of autophagy marker LC3 (second day).

(D) Fluorescent images of HeLa cells incubated with free GFP-TP53 plasmid and the plasmid mixed with Mito-FLAG (200 μM, 24 h).

(E) Cell viability of HeLa, Saos-2, and HS-5 cells incubated with free GFP-TP53 plasmid (5 μg/mL), Mito-FLAG (200 μM), and the plasmid (5 μg/mL) mixed with Mito-FLAG (200 μM, third day). Data are presented as mean ± standard deviation.

(F and G) Apoptosis analysis of HeLa (F) and HS-5 (G) cells incubated with GFP-TP53 plasmid (5 μg/mL) mixed by Mito-FLAG (200 μM, third day) via PI staining.