Figure 2
Effect of GSNO on DMT1 levels in parkin-overexpressing SH-SY5Y cells. (a) Changes of SNO-parkin and DMT1 protein levels in SH-SY5Y cells transiently expressing parkin with 200 μM of GSNO or GSH treatment for 24 h. (b) Statistical analysis of changes in SNO-parkin levels. Data were presented as the ratio of SNO-parkin/parkin. ***P = 0.0002, parkin-GSNO versus parkin-GSH. (c) Statistical analysis of changes in DMT1 levels. Data were presented as the ratio of DMT1/β-actin. &P = 0.013, parkin-GSH versus empty vector-GSH; ***P = 0.0003, parkin-GSNO versus parkin-GSH; *P = 0.025, parkin-GSNO versus empty vector-GSNO. N = 9. (d) DMT1 protein half-life (t1/2) was determined by pulse-chase assay and immunoblotting. CHX was administered in SH-SY5Y cells with different treatment to block protein synthesis. (e) Statistical analysis of DMT1 protein half-life (t1/2). P = 0.027, parkin-GSH versus empty vector-GSH; P = 0.034, parkin-GSNO versus empty vector-GSNO; P = 0.121, empty vector-GSNO versus empty vector-GSH; P = 0.074, parkin-GSNO versus parkin-GSH. N = 6.
