Fig. 1. The hsp70:^gfpsema3a1^myc transgene is heat inducible in transgenic embryos. (A) On the right, transgenic embryos (18 hpf) after heat induction at 15 hpf exhibit ubiquitous GFP fluorescence, indicating that the transgene was induced. On the left, transgenic embryos not heat induced exhibit no GFP fluorescence. (B) After heat induction of transgenic embryos, the ^GFPSema3a1^myc fusion protein was detected by western blotting using an anti-Myc antibody. No band corresponding to the fusion protein is detectable in wild-type embryos (lane 1), nor non-heat induced transgenic embryos (lane 2). An approximately 150 kDa band corresponding to the fusion protein is labeled immediately after the heat treatment (lane 3), 1 hour (lane 4), 3 hours (lane 5), 7 hours (lane 6) and 15 hours (lane 7) after heat treatment of transgenic embryos. In lanes 3-7, heat treatment consisted of raising the water temperature from 28.5°C to 38°C over a period of 15 minutes, then holding at 38°C for 30 minutes and then decreasing to 28.5°C over the next 15 minutes in a programmable water bath.