Cell lines used for culturing and transfection were COS-7 and PC12. Transgenic mice expressing N-terminal mutant huntingtin fragments (HD-N171-N82Q) were also used. The Drosophila melanogaster yw;gmr-Q120 line was obtained from G. Jackson (UCLA) and was crossed to a w stock isogenized for the X chromosome and two major autosomes.
Primary antibodies for Western Blot included antibodies to GFP (Clontech), mTOR, phosphorylated mTOR (Ser2448), S6K1, phosphorylated S6K1 (Thr389; 1A5), 4E-BP1 and phosphorylated 4E-BP1 (Thr37) from Cell Signaling Technology; antibody to huntingtin EM48 (Chemicon); and antibody to tubulin (Sigma).
For histochemical analysis, primary antibodies included antibodies to mTOR, phosphorylated mTOR (Ser2448), 4E-BP1, phosphorylated S6 (Ser235/236; Cell Signaling Technologies), LC3 (gift from T. Yoshimori, National Institute of Genetics, Japan), huntingtin and ubiquitin (both from Chemicon).