Fig. 7

Fig. 7 (A) Western blot analysis of RPS6, p-RPS6 and Actin (loading control) in whole cell lysates of WT and tti^s450 larvae between 72–120 hpf. (B) Graphical representation of the data shown in A combined with two additional experiments (each bar represents the mean +/ SD, *p<0.05). tti^s450 larvae exhibit increased levels of p-RPS6 at 96–120 hpf and decreased levels of total RPS6 between 72–120 hpf compared to WT siblings. (C) Immunohistochemical analysis of transverse sections of tti^s450 and WT larvae at 96 hpf reveals robust p-RPS6 expression in the digestive organs. Scale bars = 50 μM. (D) The persistent expression of p-RPS6 expression in tti^s450 larvae at 96 hpf compared to WT is due entirely to up-regulated Torc1 activity as shown by the disappearance of the p-RPS6 signal when larvae are pre-treated with rapamycin. (E) Inhibiting the Tor pathway in tti^s450 larvae with rapamycin in the presence of chloroquine reduces p-RPS6 expression and at the same time increases autophagic flux as shown by the increase in LC3II level. In the graphical representation of the data, each bar represents the mean +/ SD (n = 3), *p<0.05.