Figure 5.

Autophagy protects HPMEC from morphine and Tat-mediated apoptosis. HPMEC (1.25 × 10⁴ cells / well) were plated on 96-well plates followed by pretreatment with either 3-methyladinine (3-MA) (A) or rapamycin (C) before morphine and/or Tat treatment for 3 d followed by Cell Death Detection ELISA. Alternatively, cells were transfected with ULK1 siRNA or scrambled (scrm) siRNA (B) or with ULK1 expression plasmid or empty vector (EV) (D) using HiPerfect or GeneJuice reagent respectively as per the manufacturer's instructions before treatment with morphine and/or Tat. The data are average of 3 independent experiments performed in triplicates. Mean + SD, *P < 0.05, **P < 0.01, ***P < 0.001 vs. control, #P < 0.05, ##P < 0.01, ###P < 0.001 vs. morphine, $$$P < 0.001 vs. Tat, @P < 0.05, @@@P < 0.001 vs. morphine+Tat treatment.