(A-B) DN-POLG-GFP/BFP and DN-POLG-AOX/NDI1 cells were grown in the presence or absence of doxycycline (10 ng/ml) for 12 days and cell number was assessed. Cells were supplemented with pyruvate and uridine in the presence (B) or absence (A) of aspartate (20 mM). Mean ± SEM (n=3).
(C) Western blot analysis of p-AMPK, AMPK and Tubulin in DN-POLG-AOX/NDI1 untreated or treated with doxycycline (10 ng/ml) for 3, 6 and 9 days.
(D) Quantification of the western blots assessing the phosphorylation state of AMPK in DN-POLG-AOX/NDI1. Data is represented relative to total levels of AMPK and the mean value in untreated cells. Mean ± SEM (n=3).
(E-G) Western blot analysis and quantification of the acetylation of H3K9, H3K18 and H3K27 and the total expression of H3 in DN-POLG-GFP/BFP and DN-POLG-AOX/NDI1 cells untreated or treated with doxycycline (10 ng/ml) for 9 days. Data was normalized to the mean value in untreated cells. Mean ± SEM (n=3).
* indicates significance p < .05. ** indicates significance p <0.01 throughout figure 5. See also Figure S5.