Figure 6. Senescent cells exposed to high concentration of mTOR inhibitor pp242 (1500 nM) fail to rescue themselves by forming the LC3-negative vacuoles and undergo cell death. (A) IF pictures after staining with antibodies against TOM20 and LAMP1. The mitochondria and lysosomes are segregated in the vacuoles of cells exposed to 200 nM pp242, but not to 1500 nM pp242. Nuclei stained with DAPI. (B) IF with antibodies against LAMP1 and LC3. (C) Viability of senescent ERas cells treated with 1500 nM pp242 and assayed by MTT test. (D) IF with antibodies against LAMP1 and LC3 showing the absence of the mitochondria and lysosomes-containing vacuoles after mTORC1 suppression with 200 nM of rapamycin.