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. Author manuscript; available in PMC: 2010 Jun 15.
Published in final edited form as: Free Radic Biol Med. 2009 Mar 26;46(12):1614–1625. doi: 10.1016/j.freeradbiomed.2009.03.016

Figure 9. As3+ increases GCL subunit protein levels, GCL activity and holoenzyme formation.

Figure 9

(A and B) TAMH cells were treated with As3+ (10 μM) for the time periods indicated. (A) GCLC, GCLM, and βActin protein expression were analyzed by denaturing SDS-PAGE and immunoblotting as described in the Methods section. (B) Graphical presentation of the studies described in (A). GCLC and GCLM protein expression were normalized to βActin protein levels and the data presented are averages +/− SEM of at least four experiments. (C–F) TAMH cells were preincubated in the absence or presence of cycloheximide (CHX; 1.0 μg/ml) for 1 h prior to treatment with As3+ (10 μM) for 16 h as indicated. (C and E) Cells were harvested and extracts assayed for GCL enzymatic activity as described in the Methods section. The graphical data presented are averages +/− SEM of 3–7 experiments. (D and F) GCL holoenzyme formation was analyzed by native gel electrophoresis and immunoblotting for GCLC or GCLM as described in the Methods section. *p < 0.05 and **p < 0.01, indicates a significant difference in protein expression or activity compared to untreated control. #p < 0.01, indicates a significant difference in GCL activity compared to As3+ alone.