Resveratrol Modulation of Protein Expression in parkin-Mutant Human Skin Fibroblasts: A Proteomic Approach

Figure 6

Effect of resveratrol treatment on free thiol groups (P-SH) of protein and glutathionylation in patient and control cells. Primary fibroblasts from patient (P1) and control fibroblasts (CTR) were grown as specified in Materials and Methods. As indicated, cells were incubated with 25 μM resveratrol for 24 hours (Res). The P-SH levels were measured in total cellular lysate. (a) The histogram represents the percentage changes with respect to P-SH content of CTR. (b, c) The histograms represent the percentage changes with respect to P-SH content of CTR and P1 untreated cells (UN). (a, b, c) The values are means ± SEM of different experiments (). (d) A representative image of glutathionylated proteins. Proteins of cellular lysate were loaded on 12% SDS-PAGE, under nonreducing conditions, transferred to nitrocellulose membranes, and immunoblotted with the antibody against GSH (P-SSG). Protein loading was assessed with the GAPDH antibody. (e) Densitometric analysis of PSSG proteins, normalized to GAPDH, was performed considering all bands immune-revealed by the GSH antibody, excluding the protein band marked with an asterisk. This band is not specific; it also appears in SDS-PAGE under reducing conditions (data not shown). The histograms represent the percentage changes of ADU express as P-SSG/GAPDH ratio. The values are means ± SEM of different experiments (). value determined by Student’s t-test, , , .

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