Figure 6
From: Colorectal cancer cells respond differentially to autophagy inhibition in vivo
Autophagy inhibition in SW480 increases mTORC1 and mTORC2 signaling. Rictor is strongly upregulated in ATG5 and RAB21 knockdown SW480 tumors. (A) Immunoblot analysis of phosphorylated mTOR at Serine 2448 and of total mTOR, Rictor, Raptor and Tubulin from multiple independent SW480 tumors extracted five days post-implantation in CAMs. (B) Ratio of various integrated densities of phosphorylated and total proteins to Tubulin for at least 7 tumors per cell lines; mean ± SEM. mTORC1 activity was increased in autophagy-inefficient SW480 tumors. (C) Immunoblot analysis of phosphorylated S6K1Thr389, 4E-BP1T37/46, PRAST246 AKT1/2/3 and GAPDH from multiple SW480 tumors extracted five days post-implantation in CAMs. (D) Ratio of integrated densities of phosphorylated proteins to GAPDH for all tumors; mean ± SEM (n ≥ 11). (E) Immunoblot analysis of phosphorylated NRDG1T346 and Tubulin expression from multiple SW480 tumors extracted five days post-implantation in CAMs. (F) Ratio of integrated densities of phosphorylated NRDG1 to Tubulin for all tumors; mean ± SEM (n ≥ 8). Statistical analysis was performed using Unpaired T Test followed by Holm Sidak multiple comparisons test. Full-length blots are presented in Supplementary Fig. 6.
