Figure 6. Inhibition of autophagosome formation alone does not interfere with mTORC1 signalling in A549 cells.
(A) Cell extracts from A549 cells transfected with the indicated concentration of either a scrambled siRNA (SCR) or one directed towards beclin-1 (BECN1); after 72 h, cells were lysed and samples were analysed via immunoblotting with antibodies to LC3-II, p62 (a protein that is targeted to and recycled in the autophagosome), total or phospho-rpS6 and β-actin. (B) BECN1, LC3-II, and p62 were quantified using densitometric analysis and normalised against β-actin, while the signal for rpS6 P-S240/244 was normalised against total rpS6, represented as the mean of the three biological replicates. Error bars represent ± standard error of the mean. Statistical significance was determined using unpaired Student’s t-test. Note: *P≤0.05, ****P≤0.0001.
