Research Paper Volume 5, Issue 7 pp 515—530

Figure 7. Nox2-induced autophagy plays a protective role in cell survival during oxidative stress in ECs. (A) Tet-ON and Tet-OFF MHEC were transduced with Adv-mRFP-GFP-LC3 and transfected with either si-scr or si-Nox2 as indicated. Quantification of red and green fluorophores using NIH ImageJ 1.47b demonstrate significant increase in autophagy in Tet-OFF MHEC (n=50 cells), which was abrogated by knockdown of Nox2. (B) Annexin V-FITC labeling was carried out to determine apoptotic MHEC as described in the Methods. Bar graph shows apoptotic cells as percentage of total viable cells population using three independent experiments. Autophagosome-lysosome fusion blocker chloroquine induced apoptosis in both Tet-ON and Tet-OFF MHEC. However, chloroquine-induced apoptosis was significantly higher in Tet-OFF MHEC.