Fig. 6

Effects of AZD6244, NVP-BEZ235 and IR on the induction of apoptosis and autophagy. Mean percentage of cells with hypodiploid DNA content and cellular debris in cells treated with AZD6244 (red striped columns), NVP-BEZ235 (blue columns) or a combination of both inhibitors (red and blue striped columns) 24 and 48 h after IR. DMSO treated cells (blank columns) served as controls. Cells were detached with trypsin, fixed, permeabilized, treated with RNase, stained with PI and then analyzed for fluorescence by flow cytometry. The columns display means (±SD) of hypodiploid cells and cellular debris of at least three independent experiments. Statistical significant differences are indicated as follows: # P < 0.05; ## P <0.01. Statistical significant differences between sham-irradiated and irradiated control samples are not depicted for reasons of clarity. Representative Western blot analysis of expression levels of PARP, cleaved PARP, LC3-I and LC3-II in SNB19 and A549 cells c. Cells were treated with AZD6244 and/or NVP-BEZ235 before IR with 8 Gy and whole cell lysates were prepared 24 and 48 h after IR as described previously. Protein bands were normalized to the β-actin intensity and changes in protein expression are denoted by numbers if applicable. The experiment was repeated at least three times