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. Author manuscript; available in PMC: 2012 Jan 1.
Published in final edited form as: Free Radic Biol Med. 2010 Oct 21;50(1):14–26. doi: 10.1016/j.freeradbiomed.2010.10.694

Figure 1. 4-HNE rapidly increases cellular GCL activity independent of de novo protein synthesis.

Figure 1

(A) A549 cells were treated for 30 min with the indicated concentrations of 4-HNE. (B) A549 cells were treated with 50 μM 4-HNE for the times indicated. (C and D) A549 cells were pretreated for 1 h in the absence or presence of cycloheximide (CHX; 10 μg/ml) prior to treatment with 4-HNE (50 μM) for 30 min. (A-C) GCL activity and total GSH content (GSH + GSSG) of cellular extracts were assessed by the fluorescence-based NDA assay and a modified Tietze assay, respectively (*†p<0.01, **††p<0.001). (D) GCLC, GCLM, and β-actin protein expression were analyzed by immunoblotting as described in the Materials and Methods section.