Research Paper Volume 10, Issue 11 pp 3574—3589

Suppression of mTORC1 activity in senescent Ras-transformed cells neither restores autophagy nor abrogates apoptotic death caused by inhibition of MEK/ERK kinases

Figure 3. mTORC1 inhibition induces an autophagy flux, which then terminates 24 h thereafter. (A) Western-blot analysis of autophagy markers and its modulators: LC3 I and II forms, phospho-Ulk1 (Ser757, Ser555), phospho-AMPK (Thr172) as well as ERK1,2 phosphorylation. (B) Expression of GFP-mRFP-pft-LC3 vector in senescent cells exposed to pp242 for 4, 24, 72 h visualizing formation of autophagosomes and fusion of autophagosomes with lysosomes. The nuclei stained with DAPI. Histogram presents the average number mRFP foci (autophagosomes fused with lysosomes) per cell after 4, 24, 72 h of mTORC1 inhibition.