Genetic variations (Arg5Pro and Leu6Pro) modulate the structure and activity of GPX1 and genetic risk for vitiligo

Additional Supporting Information may be found online in the supporting information tab for this article: Data S1. Supplementary materials and methods. Data S2. Supplementary References. Figure S1. (a) Possible mechanism of the NO-scavenging effect of MHY384. (b) Possible mechanism of the generation of N-salicylidenecysteine from Th2 and the conversion of NO into HNO from 1,3-thiazolidines. (c) NO scavenging activity of 1,3thiazolidines compared to MHY384. Each value was expressed as mean SEM. Statistical results of one-factor ANOVA: P < 0.001 vs. a control group, *P < 0.05, ***P < 0.001 vs. a SNP treatment group. Figure S2. NO scavenging activity of MHY384 was measured by DAF-2DA (an NOdetecting fluorescent dye) in vitro (a). SNP induced-NO was used as a negative control and carboxy-PTIO was used as a positive control. NO production was determined by changes in fluorescence intensity, which were measured every 10 min for 150 min at excitation and emission wavelengths of 485 and 530 nm, respectively. Changes in NO levels in SNP-induced B16F10 melanoma cells after MHY384 treatment (b). B16F10 cells were pretreated with MHY384 (0.5, 2, and 5 lM) or 5 lM carboxy-PTIO and then treated with 200 lM SNP for 48 h. The amount of NO produced was determined by the same method as that outlined in Figure s2 (a). Each value was expressed as mean SEM. Statistical results of one-factor ANOVA: #P < 0.05, ##P < 0.01, ###P < 0.001 vs. a control group, *P < 0.05, **P < 0.01, ***P < 0.001 vs. a SNP treatment group. Figure S3. (a) The skins of HRM2 mice were pretreated with vehicle or MHY384 on designated site based on the the experimental schedule. (b) Skin lightness was measured daily before application of the sample or vehicle. Changes in ΔL* (lightness) values of each sample were calculated as the average values after UVB exposure (day 28) minus the average baseline values before any treatments (day 1). Values represent the mean SEM of 5 experiments. Data are expressed as a percentage of UVB irradiated control. As statistical significance, data were analyzed by One-factor ANOVA followed by the Bonferroni test: P < 0.001 vs. a control group, **P < 0.01, ***P < 0.001 vs. a UVB irradiated group. Figure S4. Proposed molecular pathway underlying the NO-scavenging activity of MHY384. Possible mechanism of action of MHY384 in UVB-induced NO, leading to melanogenesis. cGMP, cyclic guanosine monophosphate: PKG, cGMP-dependent protein kinase: CREB, cAMP response element-binding protein: MITF, microphthalmiaassociated transcription factor.