Figure 7: Tsc2 KO neurons exhibit morphological changes and decreased C:Y ratio in AAF media.
Differentiated N2aC were incubated in complete or AAF media for 1 hour, were fixed, and probed with MAP2 antibodies to visualize cell bodies (A). Incubation in AAF media led to a decrease in soma diameter in Tsc2 KO but not Depdc5 KO or WT neurons (N= 50 per group; B; p<0.01). TORCAR transfected Tsc2 KO, Depdc5 KO, and WT neurons were incubated in AAF media (C, striped bars in E) or in complete media with rapamycin (D; solid bars in E). A 7.4% decrease in C:Y was observed in WT neurons incubated in AAF media (C, E; p<0.05). A 14.0% decrease (p<0.05) in C:Y was observed in Tsc2 KO neurons but no change in C:Y was observed in Depdc5 KO neurons (C, E). No significant change in C:Y was observed in WT neurons after rapamycin in complete media (D, E). Data on torin1 treated WT, Tsc2 KO, and Depdc5 KO cells can be found in Figure S9. AAF= amino acid free, rapa = rapamycin, KO = knockout. Error bars in (D), standard error of the mean. *p<0.05, ** p<0.01.
