Mycophenolic Acid Overcomes Imatinib and Nilotinib Resistance of Chronic Myeloid Leukemia Cells by Apoptosis or a Senescent-Like Cell Cycle Arrest
Figure 4
MPA induced autophagy in K562 cells. K562-S, K562-R, and K562-RN cells (2.105/mL) were grown in the presence of vehicle only or MPA (3 μg/mL) for 3 days. Six hours before the end of the incubation, samples were separated in two batches and incubated in the absence or in the presence of bafilomycin A1 (20 nM) to block the autophagic flux. Then, K562 cells were washed once in PBS and lyzed in a modified RIPA buffer for detection by Western blot of LC3B and Hsp60 (as a loading control) (a). K562-S, K562-R and K562-RN cells (2.105/mL) were infected by lentivirus coding for a shRNA anti-ATG7. Lentiviral particles were incubated for 24 h with K562 cells. Then, the cells were washed twice in PBS and grown in the presence of medium for 6 days before sorting based on GFP expression and experimental use. After 3 days, each sample was analysed for ATG7 inhibition by Western blotting (b). K562-S, K562-R, and K562-RN cells (2.105/mL) and K562-S, K562-R, and K562-RN cells (2.105/mL) deficient for ATG7 were grown in the presence of vehicle only, MPA (3 μg/mL). After 3 days, SA-β-gal-positive cells were quantified by counting 102 cells on three separate fields for each condition (c). Results show the mean of three experiments.
