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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Free Radic Biol Med. 2009 Aug 14;48(1):16–25. doi: 10.1016/j.freeradbiomed.2009.08.005

Figure 2.

Figure 2

An inactivating mutation of RDH12 associated with LCA abolishes protection against 4-HNE-induced apoptosis. Cells were transfected with the expression plasmid pcDNA3.1/HIS, expressing human RDH12 variants R161 (wild-type), R161Q (common variant), and T49M (mutant inducing LCA). Stable cell lines were established and maintained by growing the cells in complete DMEM medium containing G418 (1mg/ml). A, Equal aliquots (50 μg) of microsomal fractions were analyzed by Western-blot using anti-hRDH12 antibody (1:1000 dilution) to compare expression levels. B, Cells were treated over night (20 h) with 100 μM of 4-HNE in complete DMEM. LDH activity, released by cells with damaged membranes, was measured in medium to quantify non-viable cells. Percent cell death was calculated for each cell line, according to the manufacturer’s instructions. Percent cell death in each stable cell line was then expressed in comparison to the non-transfected cell death, set at 100%. Graph shows results of 2 independent experiments, each having 3 replicates of the same conditions and error bars denote SD.