Overexpression of human 14-3-3γ mRNA in Drosophila eye tissue using the GMR-Gal4 driver. (A) Third instar larvae imaginal discs were collected and total RNA isolated. Human 14-3-3γ specific primers were used for RT-PCR to amplify human 14-3-3γ. PCR products were elecrophoresed on agarose gels and bands detected by ethidium bromide staining. Drosophila rp49 (coding for Drosophila ribosomal protein 49) was used as a loading control for RT-PCR. (B) Imaginal discs were collected as in A and total protein extracted. Immunoblotting was used to detect 14-3-3γ protein using a pan-specific anti-14-3-3 antibody. Beta-actin was utilized as a loading control. GMRG refers to the control, GMR-Gal4 driver only (lanes 1) and A5.1 strain has the GMR-GAL4 driven human 14-3-3γ cDNA (Lanes 2).