Figure 4: Ate1 overexpression in prostate cancer cells reduces stress resistance.

(A) Representative western blot of PC3-ML cells expressing Ate1-GFP. For the construction of recombinant protein, Ate1 is represented by isoform 1 (Ate1.1), the most potent and ubiquitous splice variant of Ate1. It was fused with a C-terminal GFP and cloned into a retroviral vector pBABE-puro for low-level expression in mammalian cells, which is fully functional and not expected to cause cell death as shown by previous reports(4, 30). The expression of GFP alone was used as a control. (B) PC3-ML cells transduced with Ate1.1-GFP or GFP control were treated with various doses of H₂O₂ or Staurosporine for 12 hours, then assessed for cell viability via Calcein AM staining. Graphs showing quantification from three independent experiments. Error bars represent SEM. Statistical significance of group comparison at select data points were assessed with the Student’s t-test: ** = p<0.01.