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. 2023 Apr 6;11:1063250. doi: 10.3389/fpubh.2023.1063250

Figure 6.

Figure 6

Proliferation of astrocytes remained mainly unaffected by radiation exposure. Astrocytes were exposed to X-rays (200 kV, 15 mA) with doses up to 8 Gy and fixed for Ki-67 immunostaining after different time points. Ki-67 is an indicator of proliferating cells. (A) Exemplary images of Ki-67 (pink) immunostaining showing an unirradiated control and cells 1 h after exposure to different doses of X-rays. The nucleus was stained with DAPI (blue) for all images, resulting in blue (Ki-67 negative) or blueish-pink cell nuclei with pink Ki-67 spots (Ki-67 positive, Ki-67+). The red fluorescence in the cytoplasm is due to phalloidin-647 staining of actin. Bar: 50 μm. (B, C) Quantitative evaluation of the Ki-67+DAPI+ cells after exposure of astrocytes to X-rays. (B) Dose effect curves of Ki-67+ cells 1 h and 24 h after exposure to X-rays (n = 4). (C) Kinetics of Ki-67+ cells up to 96 h after exposure to X-rays. TNF-α was added to unirradiated samples at the time of irradiation at a concentration of 20 ng/ml as it was previously described to stimulate proliferation of astrocytes. The percentage of Ki-67+ cells remained mainly unaffected by irradiation with different doses of X-rays over a time period of 1 h to 96 h. The samples were compared via one-way ANOVA, based on a sample size n = 4 (p < 0.05). Data are shown as mean ± SD.