Fig 2: H2O2 induces Cys106-SO3− DJ-1 formation in mitochondria.A549 cells were treated with 1 mM H2O2 for indicated times and DJ-1 expression was determined by western blotting in nuclear (a), cytosolic (b), and mitochondrial (c) fractions. Densitometric quantifications are also shown. Bands detected at 23 kDa and 15 kDa in mitochondrial fractions by western blotting were analyzed by mass spectrometry analysis. The presence of DJ-1 in the band at 23 kDa (d) and Cys106-SO3− DJ-1 at 15 kDa (e) is shown. f Cys106-oxidized DJ-1 in mitochondrial fractions in A549 cells by western blotting. g Densitometric quantification is also shown. h Cys106-oxidized DJ-1 (green) in mitochondria identified by Tom20 (red) using immunocytofluorescence. Nuclei were stained with DAPI (blue). The scale bar is 5 μm. i Cys106-oxidized DJ-1 and Tom20 co-localization is shown. N = 3 replicates; *P < 0.05; **P < 0.01. Data are shown as means ± s.e.m.

Fig 3: NMR relaxation analysis of picosecond–nanosecond (ps–ns) timescale dynamics in Cys106-SO3− DJ-1.a The backbone 1H-15N amide bond vector order parameters (S2) in reduced (black) and Cys106-SO3− DJ-1 (red). S2 range from 1 for immobile residues on the ps–ns timescale to 0 for highly mobile bond vectors with no preferred orientation. Cys106-SO3− formation results in a marked lowering of S2 values in several regions of DJ-1. b The ΔS2 values calculated as: ΔS2 = S2(Cys106-SO3−)- S2(reduced) for each residue. Negative values indicate residues that become more mobile in Cys106-SO3− DJ-1 and positive values indicate residues that become less mobile. Lysine residues are circled and labeled. c The ΔS2 values onto the ribbon diagram of the DJ-1 dimer. Red values indicate increased mobility and blue values indicated decreased mobility in Cys106-SO3− DJ-1. The core of the dimer experiences a slight decrease in mobility upon formation of Cys106-SO3−, while the solvent-exposed regions have strongly increased mobility

Fig 4: Effect of BPA treatment on the ratio of oxDJ-1 (10 µg/lane)/DJ-1 (3 µg/lane). Open column, control and gray column, BPA. Values represent mean ± SD (n = 3 to 6); * p < 0.05 vs. control group.

Fig 5: Mitochondrial dysfunction in A549 cells with DJ-1 knockout.A549 cells with DJ-1 KO were generated using CRISPR-Cas9 strategy. A549 cells and DJ-1 KO A549 cells were treated with 1 mM H2O2 for indicated times to determine mtDNA amount by qPCR (a), mtDNA damage using qPCR (b), and ND1 mRNA expression by RT-PCR (c). *P < 0.05, **P < 0.01, and ***P < 0.001. $P < 0.05, $$P < 0.01, and $$$P < 0.001 compared to control A549 cell line. #P < 0.05, ##P < 0.01, and ###P < 0.001 compared to control A549 cells with DJ-1 KO. N = 3 replicates. Data are shown as means ± s.e.m.