Fig. S3

Time course of Nmu overexpression (Related to Figure 3). Larvae from a hs:Nmu_Dr /+ to WT mating were heat shocked for 1 hour at 37oC at 5 dpf and fixed at the indicated times after heat shock. ISH was then performed using an nmu-specific riboprobe on dissected larval brains. Transgenic and WT larvae were subsequently identified by PCR. Ubiquitous nmu overexpression was observed at 1 hour post-heat shock, with reduced levels at 2 and 4 hours post-heat shock. No ectopic transcript was observed at 16 and 24 hours post-heat shock. No ectopic nmu expression was observed in non-heat shocked transgenic brains (No HS, insert). Chromogenic development was stopped at the same time for all samples prior to the visualization of endogenous nmu transcript. While this data shows that ectopic nmu transcript levels are ubiquitously elevated for a few hours after heat shock, the perdurance of Nmu peptide is not known.