Quantification of PARP-1 and Caspase-3 Proteins

Cells were seeded in 6 well plates (Costar; 250000 HepG2 cells/well and 125000 A549 cells/well) 1 day before incubation with macrophages. After the incubation, proteins were extracted and PARP-1 and caspase-3 protein abundance was assessed by western blotting as described previously [13 (link)]. Primary antibodies are rabbit anti-caspase-3 antibody (Cell Signaling, #9662) and mouse anti-PARP1 antibody (BD Pharmingen, #551025). Primary antibodies mouse anti-β-actin (Sigma, #A5441) or mouse anti-α-tubulin (Sigma, # T5168) were used for normalization. IRDye 800CW-conjugated goat anti-rabbit antibody (H + L; Licor, #926-32211), IRDye 800CW-conjugated goat anti-mouse antibody (H + L; Licor, #926-32210) and IRDye 680LT-conjugated goat anti-mouse antibody (H + L; Licor, # 926–68020) were used as secondary antibodies. Quantitative analysis of fluorescence intensity was measured using the Odyssey Classic Infrared Imaging System (Licor).