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. 2021 Jan 1;12(5):1520–1530. doi: 10.7150/jca.46316

Figure 2.

Figure 2

Carbon ion radiation induced delayed DNA damage repair, cell cycle arrest, cytogenetic damage, morphological change and cell necrosis, indicating the possibility of necroptosis. (A) CNE-2 and CNE-2-RR cells DNA damage repair at 24 hours following X-ray or carbon-ion exposure demonstrated by fluorescence imaging of γ-H2AX loci. Original magnification: 200 ×. (B) Fluorescence micrographs of Hoechst 33342 stained cells showing micronuclei, buds, bridges. And induced fraction (%) of cytogenetic damage (cytome) at 24 hours and 48 hours after carbon ion irradiation. Original magnification: 200 ×. (C) Percentage of cells in the G2/M (mean ± SEM) phases at 24 and 48 hours following exposure to carbon beams. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001. (D) Light microscopic images showing typical morphological features of necroptosis (flattened and enlarged cells: marked by arrows) in carbon irradiated cells. Original magnification: 200 ×. (E) Bivariate plots of Annexin-V and PI in CNE-2 and CNE-2-RR cells at 48 hours following carbon ion irradiation (Q2: necrosis; Q3: apoptosis). (F). Mean value of fractions of carbon ion-induced apoptotic (Annexin+ cells) and necrotic (including necroptosis) (PI+) cell death at 48 hours. *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001.