Research Paper Volume 12, Issue 3 pp 2440—2452

HO-1 induced autophagy protects against IL-1 β-mediated apoptosis in human nucleus pulposus cells by inhibiting NF-κB

Figure 6. HO-1 overexpressing NPCs show elevated levels of the Beclin-1/PI3KC3 complex. (A) Western blot shows the protein levels of phospho-mTOR in NPCs transfected with Ad-HO-1 and Ad-GFP. As shown, phospho-mTOR levels are comparatively similar in both Ad-HO-1 and Ad-GFP groups. (B) Western blot analysis of HO-1, PI3KC3, Atg14, Beclin-1, P62 and LC3-II/I protein levels in NPCs cultured for 72 h, 84 h or 96 h after Ad-HO-1 transfection. (C) Immunoprecipitation assay results show the amount of Beclin-1/PI3KC3 complex in HO-1 overexpressing NPCs compared with controls. Briefly, the cell lysates were immunoprecipitated (IP) with the anti-Beclin-1 antibody and the immunoprecipitated proteins were analyzed by western blotting using the anti-PI3KC3 antibody. Note: Data are represented as mean ± SD of three independent experiments; ****p<0.0001.