FIGURE 4.
Archazolid induces BNIP3 expression. A, BNIP3 protein expression was analyzed in SKBR3 cells after different durations of archazolid (10 nm) treatment. B and C, BNIP3 induced by archazolid is found in the mitochondrial fraction (B) analyzed via Western blotting after cell fractionation or (C) confocal microscopy. Mitochondria were stained by MitoTracker (green) and BNIP3 by antibody (red). D, heterodimerization of BNIP3 with Bcl-XL is shown by immunoprecipitation (IP: BNIP3) after 24 h of archazolid treatment. E, BNIP3 was silenced using siRNA 24 h before archazolid treatment for 48 h , and apoptosis induction was determined by propidium iodide staining and flow cytometry. F, cells were treated with 10 nm archazolid for 24 h, and heterodimerization of beclin-1 with Bcl-XL was determined by immunoprecipitation IP, beclin-1. One representative experiment of three is shown. At least three independent experiments were performed. *, p ≤ 0.05, Student's t test.