Fig. 2

Silibinin triggered mitochondrial ROS derived from NOX4 in PC-3 cells. a NOX isoforms were detected by PCR using specific primers for NOX isoforms. GAPDH was used as a loading control. b NOX4 expression was analyzed after treatment with 150 μM silibinin for 24 h with the presence or absence of 0.5 μM DPI. c Representative images obtained by confocal fluorescence microscopy with MitoSOX after exposure to silibinin for 24 h. After fixation and permeabilization, it was cell stained with our Nox4 antibody and appropriate FITC-conjugated secondary antibody. Nuclei were counter-stained with DAPI