Fig. 3: Lon knockdown flies exhibit increased mitochondrial transcript abundance and decreased mitochondrial translation. | Cell Death Discovery

Fig. 3: Lon knockdown flies exhibit increased mitochondrial transcript abundance and decreased mitochondrial translation.

From: Lon protease inactivation in Drosophila causes unfolded protein stress and inhibition of mitochondrial translation

Fig. 3

a Mitochondrial DNA abundance was compared in LonKD and control flies using qPCR to compare the ratio of mtDNA-encoded mt:Cyt-b to that of nuclear-encoded Act79b (n = 3 independent groups of 40–45 fly heads). Statistical significance was determined using Student’s t-test. b qRT-PCR was used to quantify steady-state abundance of the indicated mitochondrial RNAs in 21-day-old adult fly heads. Mitochondrial RNA abundance was normalized to the abundance of the nuclear-encoded Act79b transcript (n = 3 independent groups of 40–45 fly heads). Error bars indicate mean ± SEM. Student’s t-test was applied, *p < 0.05, **p < 0.005, ***p < 0.0005. c Immunoblot analysis of third instar larvae to confirm knockdown of Lon using actin as a loading control. n = 3 independent groups of five third instar larvae. Significance was determined using Student’s t-test, **p < 0.005. d In organello translation was performed using mitochondria isolated from third instar larvae. Mitochondria were labeled by incubating with 35S-methionine for 1 h. Positions of individual mitochondrially encoded proteins are indicated (left panel). Coomassie-stained gel (right panel) was used as a loading control. Control = UAS-mCherry-RNAi driven by elav-GAL4 and da-GAL4. LonKD = UAS-Lon-RNAi-1 driven by elav-GAL4 and da-GAL4

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