Fig. 2.
SCN−influences the extent and nature of HOCl-induced thiol oxidation in J774A.1 cells. Thiol concentration was assessed by the ThioGlo-1 assay after treatment of the J774A.1 cells (0.5 × 106 cells/well) with (A) HOSCN (0–200 μM), (B) co-treatment with HOCl (100 μM) and SCN− (0–200 μM) or (C) co-treatment with HOCl (200 μM) and SCN− (0–200 μM) for 1 h at 37 °C, followed by re-incubation in cell media for 0 h (black bars), 4 h (yellow bars) and 24 h (blue bars). (D) shows the GSH concentration in the cells after treatment with HOSCN (200 μM), HOCl (100 or 200 μM) or co-treatment of HOCl (100 or 200 μM) and SCN− (200 μM) for 1 h before cell lysis and derivatization of GSH with monobromobimane and HPLC separation as described above. Cellular thiols and GSH are normalized by total protein and shown as fold changes compared to the non-treated control groups. * shows a significant difference (p < 0.05) compared to the non-treated control groups with same re-incubation time period; # shows a significant difference (p < 0.05) compared to HOCl group without addition of SCN−; Δ shows a significant difference (p < 0.05) compared to counterparts (with same treatments) without re-incubation by 2-way ANOVA with a Tukey's multiple comparison test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)