Figure 5.

Oxidative stress and disturbance of autophagy flow after LED exposure. Male Wistar rats aged 7 weeks (n = 4) were exposed to white LED for 4.75 hrs and 18 hrs. NE: Non‐exposed rats. (A) At the end of the exposure period, the RPE was flat‐mounted and immunostained with anti‐p62. The RPE was further analysed by double staining with DAPI by confocal microscopy. The pictures were taken on the upper retina 100 μm away from the optic nerve. Scale bar represents 20 μm. The white arrows point p62 cytoplasmic clusters (4.75 hrs) and nuclear localization (18 hrs). (B) After 4.75 hrs (4.14 J/cm²) of light exposure the eyes were enucleated, the RPE dissected, extracted with M‐PER buffer and loaded on the top of a 10% SDS–PAGE, transferred onto a nitrocellulose membrane and probed for anti‐p62 (upper panel) and anti‐actin antibody as a charge control (lower panel). (C) The histogram shows a quantification of p62 expressed as a ratio to actin. Values are significantly different ***=P < 0.005.