Research Paper Volume 16, Issue 11 pp 9727—9752

Figure 8. Pharmacological inhibition of the HR pathway abolishes 14-3-3σ-mediated carbon ion irradiation resistance. Colony formation assay (A, B), EdU assay (C, D), and immunofluorescence (IF) staining of γH2AX 1 h (E, F) in MiaPaCa-2 cells after 2 Gy carbon ion irradiation in different groups: vector group (empty vector), 14-3-3σ group (14-3-3σ overexpression), KU-55933 group (14-3-3σ overexpression + KU-55933 [inhibitor of ATM, 10μM, dissolved in DMSO]), and VE-821 group (14-3-3σ overexpression + VE-821 [inhibitor of ATR, 10μM, dissolved in DMSO]). Colony formation assay (G, H), EdU assay (I, J), and IF staining of γH2AX 1 h (K, L) in AsPC-1 cells after 2 Gy carbon ion irradiation in different groups: NC group (negative control), si14-3-3σ group (14-3-3σ knockdown), KU-55933 group (NC + KU-55933 [inhibitor of ATM, 10μM, dissolved in DMSO]), and VE-821 group (NC + VE-821 [inhibitor of ATR, 10μM, dissolved in DMSO]). For IF, at least 100 cells per condition were analyzed. Data were presented as the mean ± standard deviation (SD) (n=3); *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001, ns: not significant. For EdU, scale bar = 100 μm; For IF, scale bar = 10 μm.