Apoptosis is not detected in young flies but is in older <i>spag</i> RNAi and wild type flies.

<p>(A) Fly heads were harvested from 7-day old or (B) 30-day old wild type flies Canton S flies (C.S.) or flies expressing the indicated transgene with the <i>tim</i>GAL4 driver at various times indicated and immunoblotted for Diap1, active Dronc or actin. Only 30-day old flies with reduced <i>spag</i> or <i>dbt</i> activity produced cleaved Diap1 (indicated by *), and <i>spag</i> RNAi flies produced active Dronc at all times at 30 days. (C) Fly heads from C.S. flies were collected at 7 and 60 days and immunoblotted for Dbt, active Dronc, Diap1 and actin. 60 day old C.S. flies showed Dbt mobility shift, active Dronc expression and Diap1 reduction at ZT7 and 19. Note that there were no <i>spag</i> RNAi flies still alive at this time. (D) S2 cells treated with <i>spag</i>, <i>dbt</i>, <i>diap1</i> dsRNA or stably expressing Dbt^K/R were immunoblotted for Diap1, demonstrating reduced Diap1 only with dsRNAi for <i>diap1</i>.