Fig 2: HDAC1 modulates OGG1 activity.a, b Representative images and quantification of OGG1 acetylation assessed by western blot analysis using antibody against acetylated lysine (Ac-Lysine, 4G12). Recombinant OGG1 (100 ng) was first acetylated by p300 (25 ng), and then incubated with titrated amount of HDAC1 (25 and 50 ng). a, b Four replicates from four independent experiments. c Reaction scheme of OGG1 cleavage assay. 5'IRDye800CW-labeled oligonucleotides (33 nt) that contain an 8-oxoG base at position 17 and complementary unlabeled strand were used as OGG1 substrate. Recombinant OGG1 proteins were incubated with OGG1 substrate and resolved by 20% denaturing urea PAGE. d, e Representative images and quantification of OGG1 cleavage activity on the reaction samples shown in Fig. 3a. d, e Seven replicates from four independent experiments. Nuclear extracts from HEK cells overexpressing OGG1 were used as a positive control. f Co-immunoprecipitation of OGG1 with HDAC1 in cortical nuclear extracts from 2-month-old wild-type mice (n = 2) from one experiment. The asterisks indicate the nonspecific cross-reaction with the IgG heavy and light chains. g, h Representative images and quantification of acetylation status of endogenous OGG1 in hippocampal lysates of 13M control (n = 4) and Hdac1 cKO (n = 4) animals. g, h Two independent experiments. i Representative images of OGG1 cleavage assay using hippocampal nuclear extracts of 13M control and Hdac1 cKO mice. j Quantification of cleaved 17-nt products after OGG1 cleavage assay using hippocampal nuclear extracts of 13M control (n = 6) and Hdac1 cKO (n = 4) animals. i, j Two independent experiments. All values are shown as mean ± SEM. Statistical analysis: b, e one-way ANOVA with Tukey’s post hoc test; h, j two-tailed Student’s t test. Source data are provided as a Source Data file.

Fig 3: Imunnohistochemistry photomicrographs. (A) Strong intensity cytoplasmic and nuclear immunostaining of SOD-1. (B) Low intensity cytoplasmic immunostaining of SOD-1. (C) Strong intensity cytoplasmic and nuclear immunostaining of OGG1/2. (D) Low intensity cytoplasmic immunostaining of OGG1/2. (E) Strong intensity cytoplasmic immunostaining of Trx. (F) Low intensity cytoplasmic immunostaining of Trx. (G) Strong intensity cytoplasmic and nuclear immunostaining of Ref-1. (H) Low intensity cytoplasmic immunostaining of Ref-1. SOD-1 superoxide dismutase, Ref-1 purinic/apyrimidinic endonuclease/redox factor-1, OGG1/2 8-oxoguanine glycosylase. Original magnifications of ×400.

Fig 4: MTH1 deficiency reduces the expression of mtDNA-encoded genes after thrombin stimulation.a Western blot analysis of the expression of MUTYH, OGG1/2, MTH2, and MTH3 in MTH1fl/fl and MTH1-/- platelets under resting condition. The representative images were shown from three independent experiments (mean ± SD, n = 3 independent isolated platelets). b Mice were administered a Dylight 488-labeled anti-GP1bß antibody (Emfret, X488) via tail vein (0.1 µg/g body weight) to measure platelet half-life by flow cytometry. Data were presented as mean ± SE (n = 3, two-way ANOVA with Sidak multiple comparisons test. c Expression of subunit of complexes (CI, CIII, CIV and CV) in the mitochondrial respiration chain proteins encoded by mtDNA (ND1, CYTB, MT-CO1, ATP8) or nucleus DNA (NDUFV1, UQCRC2, COX6A1, ATP5A) in platelets from MTH1fl/fl or MTH1-/- mice before (-) and after (+) stimulation with thrombin (1 U/ml) for 3 mins (representative of three independent experiments) (mean ± SD, n = 3, two-way ANOVA with Sidak multiple comparisons test). d MT-CO1 gene expression level in thrombin-activated platelets from MTH1fl/fl or MTH1-/- mice was measured by quantitative real-time PCR and represented as a fold change relative to its level in resting platelets (mean ± SD, n = 3 independent isolated platelets, two-tailed unpaired Student’s t test). e Analysis of the number of guanine in the 13 mtDNA sequences.