Figure 4.

FdR cells are addicted to basal autophagy. (a) Western blot analysis for the levels of LC3 I/II and β-actin used as a loading control, in Nalm-6 and their derivative FdR cells in response to fludarabine (Fd)-treatment for the indicated time. (b) Nalm-6 and Nalm-6 FdR cells were pretreated with CQ for 1 h ± Fd (4 h). Cells were then fixed, stained with an antibody against LC3 and LAMP2 along with nuclear staining by DAPI and analyzed by immunofluorescence. (c) Correlation index for LC3 and LAMP2 in the indicated samples. (d) Cell death analysis in ShControl and shLAMP2-expressing Nalm-6 FdR cells in response to 24 h Fd treatment, as determined by trypan blue staining. (e) Western blot analysis for the levels of LAMP2, as well as β-actin in Nalm-6 FdR cells expressing shControl or shLAMP2. (f) Nalm-6 FdR cells were pretreated with CQ for 1 h followed by 24 h Fd treatment and cell death was determined by AnnexinV/PI staining. (g) Dose reponse for the effect of 24 h Fd treatment on cell growth in Mec-2 cells, as determined by the MTS assay. (h) Mec-2 cells were pretreated with CQ for 1 h followed by 24 h Fd treatment and cell death was determined by AnnexinV/PI staining. All data are expressed as mean±S.D. of percentage of cell death from three independent experiments; *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n=3