The UPR ^mt enhances the transcription of mitochondrial import machinery. (a) To induce UPR^mt during development, glp-1(e2141ts) mutant animals were grown at 25°C on bacteria expressing cco-1 dsRNA from the time of hatching until the first day of adulthood (animals were treated with 1:1 mixture of bacteria containing the empty RNAi vector alone [EV] to match with the double RNAi treatment). To suppress UPR^mt, animals were treated with double RNAi (1:1 mixture of bacteria replacing EV with bacteria expressing dve-1 dsRNA). Control worms were grown on bacteria containing EV alone. RNA was isolated on day 1 of adulthood, and qPCR analysis was performed. Expression was normalized against three housekeeping genes and quantified with Student’s t test (two-tailed). The graph is presented as mean ± SD, n = 2. *, P < 0.05; **, P < 0.01. (b and c)glp-1 animals were grown at 25°C on bacteria expressing timm-23, cco-1, or daf-2 dsRNA (each was diluted to 1:1 ratio with bacteria containing the empty RNAi vector alone) from hatching until the first day of adulthood. Control worms were grown on bacteria containing the empty vector alone. Quantification is shown in c with unpaired Student’s t test (two-tailed). n = 3. **, P < 0.01. The signal intensity of TIMM-23 was normalized to that of NDUFS3. (d and e)hsp-6p::gfp animals were grown at 20°C and treated with RNAi or empty RNAi vector control from hatching until the L4 stage, then transferred to plates of the same RNAi treatment with the addition of TMRE and grown overnight. Bacteria expressing cco-1 dsRNA (right) or mrps-5 dsRNA (middle; both were diluted 20% with bacteria containing the empty RNAi vector alone) were used to induced UPR^mt. Control worms were grown on bacteria containing the RNAi vector alone (left). Scale bar = 50 μm. Fluorescent intensity was quantified with ImageJ and analyzed with unpaired Student’s t test (two-tailed). n = 3. *, P < 0.05. All graphs are presented as mean ± SD. Source data are available for this figure: SourceData F4.