Fig. 2.

Nuclear DNA leakage activates nucleophagy. (a and b) MDA-MB-231 cells were exposed to various concentrations of DOX for 24 h, or treated with DOX (10 μM) for different times. The expressions of autophagy-related proteins, LC3B-I/LC3B-II and SQSTM1, in whole cellular (WCL) and nuclear extracts (nucleus) were detected by western blot. (c) MDA-MB-231 cells transfected with EGFP-LC3 were treated with DOX (10 μM) for 24 h. Cells were stained with DAPI and the colocalization of EGFP-LC3 (green) and leaked DNA (blue) was examined by confocal microscopy. Scale bars: 10 μm. (d) Confocal microscopy images of MDA-MB-231 cells treated without or with DOX (10 μM) for 24 h after co-expression of mRFP-LC3 (red) and mGFP-LAMP1 (green) and staining with DAPI (blue). Scale bars: 10 μm