Fig. 2

OXR1 regulates cell proliferation, anti-oxidation response and apoptosis. a Proliferation capacity of patient (OXR^ΔEx18) and control (CTRL) lymphoblasts. n = 3 independent experiments, n = 6 samples for each genotype in each experiment. b OXR1^ΔEx18 and CTRL lymphoblasts were treated with indicated concentrations of hydrogen peroxide (H₂O₂) for 1 h and harvested after 24 h recovery. Populations of surviving (left), apoptotic (middle) and necrotic (right) cells were determined by flow cytometry with FITC-Annexin V/PI double staining. n = 3 independent samples. c The detection of 8-oxoG levels in genomic DNA by mass spectrometry after H₂O₂ treatment in OXR1^ΔEx18 and CTRL lymphoblasts. n = 3 independent samples in the first assay (0.25 mM), and n = 6 independent samples in the second assay. d Western blot analysis and quantification of p21 and HO-1 expression under regular culturing conditions in OXR1^ΔEx18 and CTRL lymphoblasts. n = 3 independent samples. e Quantitative real-time PCR analysis of oxidative stress response genes in OXR1^Ex18 and CTRL lymphoblasts exposed to H₂O₂ and recovery for 6 h (R6h). n = 3 independent samples. f qPCR analysis of Caspase-9 (CAS9) expression in OXR1^ΔEx18 and CTRL lymphoblasts following 0.25 mM H₂O₂ treatment and recovery for 6 h (R6h). n = 3 independent samples. g Western blot analysis and quantification of CAS9 expression and cleaved CAS9 (cCAS9) in OXR1^ΔEx18 and CTRL lymphoblasts exposed to 0.25 mM H₂O₂ and recovery for 3 h (R3h) or non-treated (NT). β-ACTIN used as loading control, n = 3 independent samples. h Proliferation capacity of patient lymphoblasts with stable expression of OXR1D (LB-OXR1^ΔEx18 + pE-OXR1D) compared to patient lymphoblasts carrying only vector (LB-OXR1^ΔEx18 + vector), n = 6 independent samples. i Analyses of cell viability, apoptosis and necrosis patient lymphoblast samples stably expressing OXR1D (LB-OXR1^ΔEx18 + pE-OXR1D) or empty vector (LB-OXR1^ΔEx18 + vector) upon H₂O₂ treatment. n = 3 independent samples. All Data are shown as mean ± SD. *p < 0.05, **p < 0.01, multiple t test using the Holm-Sidak method to correct multiple comparisons when determining the statistical significance