Fig. 3

N-7DHC-lipos plus irradiation induces lipid peroxidation, tested with H1299 cells. N-7DHC-lipos were applied at a dose of 50 µg/mL (an equivalent 7DHC dose of 5 µg/mL), and irradiation (IR) was applied at 5 Gy (n = 3 biologically independent samples). (a, b) Influence on cellular superoxide (a) and hydroxyl radical (b) levels, measured by DHE and APF, respectively. (c) Influence on cellular SOD activity. (d, e) Cellular superoxide (d) and hydroxyl radicals (e) upon treatment with N-7DHC-lipos plus irradiation while co-incubated with nystatin (NY), tocopherol (TP), or ascorbic acid (AA), measured by DHE and APF, respectively. (f) Double-strand breaks, measured by flow cytometry with anti-γH2AX-stained cells. (g) Induction of apoptosis, measured by Western blotting analyzing the levels of c-PAPR and c-Caspase3. (h-j) Influence on lipid peroxidation, measured by C11-BIDOPY (h), 4-HNE (i), and TBARS (j) assays. (k) Intercellular sterol and oxysterol levels, measured by UPLC-MS/MS (n = 3 biologically independent samples). Data in the heat map are z-scored. Data are presented as mean ± SD. Statistical difference was evaluated using one-way ANOVA in (a-f, and h-j). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001