Figure 7.

JNK is not involved in taxol-induced autophagy activation. MDA-MB-231 cells were untransfected (X) or transfected with 25 nM of JNK1 and 25 nM of JNK2 siRNA (SI) or negative control RF siRNA at 50 nM (RF) for 24 h. The transfection media were removed and replaced by culture media for 24 h. Cells were then incubated under normoxia (N) or hypoxia (H) for 16 h, without (C) or with taxol (T) at 50 μM. (a) LC3 and p62 were detected in total cell extracts, obtained with a phospho-protein-specific lysis buffer, by western blotting analysis, using specific antibodies. β-actin was used to assess the total amount of proteins loaded on the gel. (b) Cells were incubated in the presence of the DQ-Green-BSA fluorescent dye (unlabeled cells serve as a negative control). After incubation, cells were harvested and analyzed by flow cytometry