Increased phosphorylated S6K is observed in fed buffy mutant larvae. (A) A representative immunoblot demonstrating that 4E-binding protein (4E-BP) protein is detected in protein-starved wild-type and buffy larvae. (B) More phosphorylated S6K is detected in fed buffyH37 mutant larval extracts relative to wild-type and target of rapamycin (Tor) signaling is downregulated by starvation in both genotypes. Data shown is from successive incubation with the indicated antibodies on the same western blot. (C) Graphical representation of fold increase of phosphorylated S6K in buffy mutants relative to wild-type (= 1, dashed line) for five separate biological experiments with the mean indicated by the horizontal bar.