Fig. 1.

MT-ND5 editing window, editing efficiency, and cellular tolerance to the induced mutations. A) DdCBE editing window demonstration. Only C4 and C7 were edited within the editing window. B) Peak heteroplasmy across cell lines. C) Temporal changes in heteroplasmy in HEK293. HEK293T cells display a pattern similar to that of HEK293. D) Sanger sequencing trace of editing window on the template strand in HEK293 cells, illustrating a C:T transition in the sense strand. This corresponds to a G:A transition in the displayed template strand. Arrows indicate positions C4 and C7. E) Temporal changes in heteroplasmy in MCF12A. F) Sanger sequencing trace for MCF12A. G) Temporal heteroplasmy change post retransfection for C4 (left panel) and C7 (right panel), for the c2 and n groups. H) Supplements improve heteroplasmy. ‘no sup’, no supplements control group; ‘sup-U’: supplemented with sodium pyruvate, glutamine, and non-essential amino acids; ‘sup + U’: sodium pyruvate, glutamine, nonessential amino acids, and uridine. This assay was conducted in MCF12A cells at early stage heteroplasmy (6 days post-transfection). The Jonckheere-Terpstra test was used to assess trends in heteroplasmy under different treatments. Boxplots represent the median and interquartile ranges. Heteroplasmy level data in B), C), E), and G) represent the mean±sem of technical triplicates.