Fig 4.

Neuronal dCbl knockdown results in upregulated dilp gene expression in parallel with increased phosphorylation of dAkt and dERK. (A) RNA was extracted from the head of fed or starved (Stv; 24 h) male adult elavG4>+ or elavG4>dCbl-Ri flies (3 days of age, 90 flies/group). The abundance of dilp2, dilp3, or dilp5 mRNA was measured by qRT-PCR (n > 3 independent experiments). After normalization to the levels for RPL32, results are shown as means ± SEMs; *, P < 0.05 versus the value from control elavG4>+ flies by one-way ANOVA. (B) Western immunoblot analysis of dAkt and dERK phosphorylation using the indicated antibodies. Head or body protein extracts were analyzed for fed or starved (24 h) male adult flies (5 days of age, 90 flies/group). Representative results from three independent experiments are shown. Drosophila α-tubulin was used as an internal control. (C and D) Relative phosphorylation levels of dAkt (C) and dERK (D) were determined by densitometric quantification of the immunoblots, presented as the mean ± SEM (n = 3). *, P < 0.05 versus control elavG4>+ flies; #, P < 0.05 between fed and starved conditions by two-way ANOVA.