After a 10 minute baseline in complete media WT, scramble, Tsc2 KO, and Depdc5 KO cells transfected with TORCAR were incubated in complete media containing torin1 (A-D, 100 nM, mins 10–30, solid bars in E) followed by incubation in AAF media containing torin1 (100 nM, mins 30–60, striped bars in E). During incubation in complete media containing torin1, an average decrease in C:Y was observed across each experimental group: WT = 3.8%, Scram = 3.0%, Tsc2 = 3.7%, Depdc5 = 2.2% (p <0.05). After incubation in AAF media containing torin1, similar percent decreases in C:Y ratio were observed in WT (A; 4.9%), scramble (B; 3.2%), Tsc2 KO (C; 3.4%) and in Depdc5 KO N2aC (D); 1.7%) (p <0.05); Results were statistically significant from baseline but not between groups (e.g., torin1 in complete media vs. torin1 in AAF media).