(a) Comparison of body weight between different
DmPOLγA compound-heterozygous larvae. All
DmPOLγA compound heterozygous larvae had wild
type-like body size. Body weight of twenty 5-day-old larvae was measured and
average body weight is shown. Tukey’s Multiple Comparison Test.
***p<0.001, ** p<0.01, *p<0.05. Error bars represent S.D.
n=20.
(b) Quantification of unique mtDNA mutations in compound
heterozygous larvae. The homozygous exo- larvae had significantly more unique
mtDNA mutations in comparison with compound heterozygous larvae. Both genotypes
inherited mtDNA mutations maternally for one generation. Tukey’s Multiple
Comparison test. ***p<0.001, ** p<0.01, *p<0.05. Error bars
represent S.D. n=3.
(c) Quantification of unique mtDNA mutations in adult compound
heterozygous flies without and with maternally transmitted mutations. Compound
heterozygous flies with maternally transmitted D263A exo- allele (black bar)
showed increase in the number of unique mtDNA mutations as compared to compound
heterozygous flies that inherited the exo- allele paternally (grey bar). One-way
ANOVA with Dunnett’s post hoc test. ***p<0.001, ** p<0.01,
*p<0.05. Error bars represent S.D. n=3-6.
(d/e) An increase in the mtDNA mutation load affects fly
development. Developmental time of different complementation groups is shown.
(d) All genomically engineered flies with a clean background
(lacking maternally transmitted mtDNA mutations, left panel) had the same
developmental time as wild-type (WT) flies. In contrast, all flies inheriting
mtDNA mutations maternally for 4 generations (D263A/Rescue, D263A/H1038A and
D263A/Q1009A) showed developmental delay (right panel). Crossing schemes are
shown on Supplementary Fig.
7a. Data represent two independent experiments. (e)
Wild-type (WT) flies carrying mtDNA mutations showed a severe developmental
delay. Compound heterozygous flies used in (d) were outcrossed
twice in order to replace tamas mutant alleles with wild type
tamas allele and clean the nuclear background. All flies
had a WT nuclear background with or without maternally inherited mtDNA
mutations.