Schematic representation of the viral vectors and Western blot analysis of Akt expression. A. Diagram of RCASBP(A), RCASBP(A) KRas, RCASBP(A)Tet-off, RCANBP(A), RCANBP(A) TRE-Akt. These vectors are all Gateway compatible to allow for the easy insertion of experimental sequences. LTR, long terminal repeat; Y, packaging signal; SD, splice donor; SA, splice acceptor. B. Nestin-TVA-positive astrocytes were infected in culture with RCASBP(A)Tet-off alone (lanes 1 and 2), RCANBP(A)TRE-Akt alone (lanes 3 and 4) or RCANBP (A)TRE-Akt with RCASBP(A) Tet-off (lanes 5 and 6). The samples in lanes 2, 4, and 6 were treated with 2 μg/ml doxycycline for 48 h. Lys-ates were separated on a 10% Tris-glycine gel, transferred to nitrocellulose, and probed with an antibody against the HA epitope tag on Akt. The membrane was re-probed with an antibody against α-tubulin to ensure equal loading.