Age of onset and time course of mortality in ND23⁶⁰¹¹⁴ flies. (A) Percent mortality was determined for mixed sex ND23⁶⁰¹¹⁴ flies not exposed (control) or exposed to 2 h of 2% isoflurane and 21% O₂ at the indicated ages. Significance between control and experimental data was determined using the unpaired equal-variance two-tail Student’s t test. ****P < 0.0001. (B) The time at which 10- to 13-day-old, mixed-sex ND23⁶⁰¹¹⁴ flies died during the 24 h after exposure to 2 h of 2% isoflurane and 21% O₂. At 2-h intervals the percentage of dead flies was normalized to the percentage of flies that were dead at 24 h. (C) 10- to 15-day-old, mixed-sex ND23⁶⁰¹¹⁴, ND23⁶⁰¹¹⁴/ND23^G14097, and ND23⁶⁰¹¹⁴/ND23^Del flies were exposed to 2 h of 2% isoflurane and 21% O₂, and the percent mortality at 24 h was determined. N = number of biologic replicates. Symbols indicate the following: box, second and third quartiles of data; +, mean; horizontal bar, median; and whiskers, (minimum and maximum.