Fig. 7

Everolimus modifies autophagic cell markers. a Inhibition of mTOR phosphorylation, downregulation of HSP70 and HSP27 as well as induction of PARP cleavage by 20 nM everolimus are shown by western blot. b MCF-7, MCF-7:5C and MCF-7:2A cells were treated with vehicle, 20 nM everolimus, 50 μM chloroquine (CQ) or both for 24 h. LC3B is shown by immunofluorescent confocal microscopy. Punctate LC3B granules (green) are indicated with white arrows. Cell nuclei are stained with DAPI (blue). Mean fluorescent intensity was determined using Image J. c Cells were treated with vehicle, 20 nM everolimus, 50 μM chloroquine (CQ) or both for 48 h and LC3-I and -II assessed by western blot. The ratio of LC3-II/β-Tubulin normalized to control was determined by densitometry with Image J and is shown below the blots