Fig. 3. NOXA gets degraded at lysosomes.

Western blot analysis of HeLa cell lysates a transfected with HA-NOXA lysine mutants (KxxxR) as indicated for 36 h, b treated with 100 nM BAF, 10 µM CQ and 30 mM NH₄Cl for 16 h, as indicated. c Western blot analysis of cell lysate (Input) and Strep-PD of HEK293T cells transfected with Strep-Ub K63 only (CTRL) for 36 h and treated with 100 nM BAF or 5 µM MG132 for 16 h. a–c Actin served as a loading control. d Confocal microscopy of HeLa cells transfected with GFP-NOXA^wt or GFP-tBID for 16 h and mitochondria were stained with αTom20 antibody (568 nm) or co-transfected with mCh-Mito construct. Nuclei were counterstained with DAPI. e STED microscopy of HeLa cells transfected with GFP-NOXA^wt or GFP-tBID for 16 h. f Confocal microscopy of HeLa cells transfected with GFP-NOXA^wt and Lamp-1-mCh for 16 h. NT not transfected. DOX doxorubicin. CQ chloroquine. BAF bafilomycin. MG MG132. mCh mCherry. Mito Mitochondria. All experiments are representatives of at least three independent experiments.