JY-1-106 induces apoptosis in Mcl-1 overexpressed cancer cells. (A) Multiple cells lines grown under regular culture conditions were collected and lysed. Bcl-xL and Mcl-1 expression in these cell lines were measured via western blotting. The β-actin expression levels were detected to normalize for protein loading. (B) To determine the response of A549, DLD-1, DLD-1BR, H1299, H23, I45, I45BR and REN cells to various chemotherapeutics including ABT-737, JY-1-106, cisplatin and SAHA, these cells were plated at a density of 3000 cells per well and treated with different concentrations of these drugs over a 72 hour period. An XTT assay was then performed in quadruplicate for each treatment condition. (C) REN and DLD-1 BR cells were transfected with control siRNA or Mcl-1 siRNA via electroporation. The cells were then seeded in 96 well plates and exposed to various doses of ABT-737. After 72 hours in culture, an XTT assay was performed to evaluate cell viability. This experiment was repeated three times. The differences between these two groups were measured using the Student’s t-test (p < 0.01). Western blotting of Mcl-1 was performed to evaluate Mcl-1 down-regulation in this experiment. (D) A549 and HMVEC cells were then seeded in 96 well plates and exposed to various doses of JY-1-106. After 72 hours of culture, an XTT assay was performed in quadruplicate for each treatment condition. The difference between A549 and HMVEC was compared using the Student’s t-test (p < 0.01). This experiment was repeated twice.