Overexpression of human 14-3-3γ in Drosophila using the Hsp70 promoter. (A) Third instar larvae harboring the human 14-3-3γ driven by the heat shock inducible Hsp70 promoter were collected after incubating at 37°C for one hour and the heat shock applied either once or multiple times as indicated in the figure. Total RNA was extracted and RT-PCR conducted using primers specific for the human 14-3-3γ RNA and the PCR products electrophoresed on agarose gels. Yw, yw67C 2, was used as a control. The experiment was repeated three times. Third instar larvae with the Hsp70 promoter driven human 14-3-3γ were collected after heat shocking at 37°C and allowed to recover as indicated in the figure. Total proteins were extracted and immunoblotted for presence of the human 14-3-3γ protein using a pan-specific anti-14-3-3 antibody (B and C). β-tubulin was used as loading controls in B) and C). The abbreviations depict the following: A) HS, heat-shock treatment; yw, yw67C 2(control); h1433GA/GC, human 14-3-3 transgenic animals (h14-3-3A+C); 1× (heat-shocked once); 2× (heat-shocked twice); 3× (heat-shocked third time); h1433γ, human 14-3-3γ primers; rp49, a loading control (Drosophila ribosomal protein encoding gene); RT, recovery time.