Fig. 4.

Regulation of larval protein synthesis by hypoxia and heat stress. (A) Third instar larvae were either maintained in room air (normoxia) or exposed to 5% O₂ (hypoxia) for 4 h. Larvae were then inverted and incubated in Schneider's media+puromycin (5 µg/ml) for 40 min. Equal amounts of whole larval protein extracts were then analyzed by western blotting. Left, western blot with anti-puromycin antibody. Right, Ponceau S staining showing total protein levels. (B) Third instar were either maintained at 25°C (control) or exposed to a 1-h 37°C heat shock. Larvae were then inverted and incubated in Schneider's media+puromycin (5 µg/ml) for 40 min. For the heat-shock samples the puromycin incubation was carried out either at room temperature (a) or at 37°C (b). Equal amounts of whole larval protein extracts were then analyzed by western blotting. Left, western blot with anti-puromycin antibody or anti-tubulin antibody. Right, Ponceau S staining showing total protein levels. All experiments were carried out using w¹¹¹⁸ larvae.