Fig. 3

HDL attenuated PA-induced ROS production and cell apoptosis. H9c2 cells were incubated with PA (0.5 mM) in the absence or presence of different concentrations of HDL (25-100 μg/ml) for 24 h. a Cell viability was determined via MTT assay. b Flow cytometry profile represents Annexin-V-FITC staining in x axis and PI in y axis. The number represents the percentage of early apoptotic cells in each condition. c Fluorescence images showed the cells stained with 4,6-diamidino-2-phenylindole (DAPI) (upper panel) and stained using terminal deoxynucleotidyl transferase dUTP-mediated nick-end labeling (TUNEL) assay (middle panel), and photomicrographs were from phase-contrast microscopy (bottom panel). d TUNEL positive cell was determined via flow cytometric analysis. e Cellular ROS was determined via MitoSOX™ (5 μM). f Fluorescence intensity of cells was measured by phase-contrast microscopy. g SOD1 and SOD2 expression was estimated by immunoblotting. h Neonatal cardiomyocytes were treated with HDL 100 μg/ml for 2 h and then incubated with 0.5 mM PA for an additional 24 h, and followed by 1 h incubation with MitoSOX™ (5 μM). Fluorescence intensity of cells was measured by phase-contrast microscopy and. Data showed the means±SEM of 3 analyses. # p < 0.05 vs. control; *p < 0.05 and **p < 0.01 vs. palmitic acid alone treatment