Fig. 2: Mettl3 knockdown animals are resilient to heat stress.

a Schematic of HS stress assay. Flies are 6-day-old. Flies are put onto RU486 food if needed to turn on the expression of the RNAi transgene for 6 days. Flies are HS at 38.5 °C and allowed to recover overnight in normal food or RU486 food at 25 °C. Percent survival in each vial of 20 animals are scored after 24 h. b Ubiquitous knockdown of Mettl3 (DaGal4, DaGS), Neuron-specific knockdown (ElavGS), and corresponding controls (mCherry RNAi) were HS for 1.5 h at 38.5 °C and scored for survival after 24 h recovery. n = 7 biological replicates, each data point represents percent survival in a vial of 20 flies per replicate. Data presented as mean ± SD, **p < 0.01, ****p < 0.0001, Student’s two-tailed t-test. p = 0.0011, p < 0.0001, p < 0.0001. c Neuron-specific (ElavGS) upregulation of Mettl3 and control were HS for 1 h at 38.5 °C and scored for survival after 24 h recovery. UAS-Mettl3 flies were severely stress-sensitive, and HS time was reduced to 1 h to have at least 5–10% of the flies surviving. Each data point represents percent survival in vials of 20 flies per replicate. Data presented as mean ± SD, ****p < 0.0001, Student’s two-tailed t-test, p < 0.0001. d Catalytic Mettl3 Mutant (ΔCat /Df) vs Mettl3 deficiency (Df/+) vs w¹¹¹⁸ (BL5905) (+/+) were HS for 1.5 h at 38.5 °C and scored for survival after 24 h recovery. Each data point represents percent survival in vials of 20 flies per replicate. Data presented as mean ± SD, **p < 0.01, ****p < 0.0001, One-way ANOVA with Tukey’s test. p < 0.0001, p < 0.0001, p = 0.0014. Source data and statistical analysis are provided as a Source Data file.