Skip to main content
. Author manuscript; available in PMC: 2013 Sep 1.
Published in final edited form as: Free Radic Biol Med. 2012 Jul 8;53(5):1111–1122. doi: 10.1016/j.freeradbiomed.2012.06.042

Fig.7. Deficiency of αB crystallin augments ER stress induced apoptosis.

Fig.7

A. RPE cells from αB crystallin (−/−) mice grown to confluence were treated with 50ng/ml and 100ng/ml TM for 24 h and stained for apoptosis by TUNEL assay (left panel). Cell lysates from RPE cells of αB crystallin (−/−) mice and control mice were probed with active caspase 3 antibody (right panel). A dose dependent increase in apoptosis and active caspase was found which was significantly higher in αB crystallin (−/−) RPE vs control RPE. B. GRP78 and αB crystallin levels as determined by western blot analysis of αB crystallin siRNA transfected RPE showed a significant decrease in αB crystallin while GRP78 remained essentially unaltered. C. Quantitation of apoptotic cell death by confocal microscopy showing increased percentage of TUNEL positive cells in αB crystallin siRNA group. (D) In the αB crystallin siRNA transfected hRPE at the maximum ER stress used (3μ g/ml TM for 24h), >90% of the dead cell population was AnnexinV+/PI indicating an apoptotic mechanism of cell death while <10% of cells were positive for both Annexin V and PI indicating either necrosis or later stage apoptosis. Values are mean ± S.D. from three independent experiments. *p<0.05.