Fig. 5

Banf1 A12T inhibits repair of oxidative lesions in NGPS patient cells. a Interactions of PARP1 with Flag-Banf1 mutants. Flag immunoprecipitations from HEK293T cells ectopically expressing the Flag-Banf1 WT or A12T proteins following H₂O₂. b Purified Banf1 WT or A12T proteins were incubated with PARP1, immunopreciptated with PARP1 antibodies and immunoblotted with the indicated antibodies. c Inhibition of auto-poly-ADP-ribose activity of PARP1 in U2OS expressing ectopic Flag-Banf1 WT or A12T following H₂O₂. d The PAR bands in (c), were analysed via densitometry and normalised to γ-Tubulin. Histogram data shown in (d), represent the mean and S.D. of three independent experiments. e In vitro inhibition of PARP1 poly-ADP-ribose activity on histones by purified Banf1 WT or A12T. f The PAR bands on (e), were analysed via densitometry. g Banf1 inhibits repair of oxidative DNA damage. Alkaline comet assay showing the relative olive tail moment in control cells and cells ectopically expressing Flag-Banf1 WT or A12T following H₂O₂ treatment and recovery. Paired t test was used for statistical analysis. h Inhibition of Poly-ADP-ribose activity of PARP1 purified from NGPS patient cells on an immobilised histone substrate HEK293T following H₂O₂. i NGPS patient cells exhibit defective repair of oxidative DNA damage. Alkaline comet assay showing the relative olive tail moment in control cells and NGPS cells after H₂O₂ treatment and recovery. Paired t test was used for statistical analysis. Immunoblots are representative of n = 3 independent experiments. Unless otherwise stated, histogram data shown represent the mean and S.D. of n = 4 independent experiments and statistical significance was defined via ANOVA. *P < 0.05, **P < 0.01. Source data are provided as a Source Data file.