Fig. 2.

Regulation of larval protein synthesis by nutrients and TOR signalling. (A) Fed or 6-h starved third instar larvae were inverted and incubated in either PBS+puromycin (5 µg/ml) or 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 either anti-puromycin, or anti-tubulin antibodies. Right, Ponceau S staining showing total protein levels. (B) Larvae were inverted and incubated in Schneider's media+puromycin (5 µg/ml) either with DMSO (control) or Rapamycin (20 nM), for 40 min. Equal amounts of whole larval protein extracts were then analyzed by western blotting. Left, western blot with either anti-puromycin, or anti-phospho-S6K antibodies. Right, Ponceau S staining showing total protein levels. All experiments were carried out using w¹¹¹⁸ larvae.