Article Figures & Data
Figures
- Fig. 1.
Induction of apoptotic cell death in the 3SB cell line and mutant sublines. a, fluorescence microscopic photographs of 3SB and 1B1C4 cells. Nuclear morphology is shown in unirradiated 3SB cells (A), in unirradiated 1B1C4 cells (B), in X-irradiated 3SB cells (C), and in X-irradiated 1B1C4 cells (D). After irradiation with 5 Gy of X-rays, the cells were incubated for 6 h and stained with Hoechst 33258. b, time course of the appearance of apoptotic cells after 5 Gy of X-ray irradiation. The percentage of apoptotic cells was calculated as the percentage of cells with abnormal nuclear morphology in 3SB cells (•) and in five radioresistant sublines (1B1C4, ○; 1A1-6, ▵; 1A3-4, ▿; 1D5-8, □; 2A1-1, ⋄). Each point represents the mean of two independent experiments.
- Fig. 2.
Western blot analysis of the expression of p21 protein in 3SB, 1B1C4, 1A1-6, and 1D5-8 cells at various times after X-ray irradiation. The cells were unirradiated (0 h) or irradiated with 5 Gy of X-rays and harvested 1.5, 3.0, 4.5, and 6.0 h after exposure.
- Fig. 3.
NF-κB DNA binding activity and the expression of NF-κB and IκBα in X-irradiated thymic lymphoma cells. a, changes in NF-κB DNA binding activity in 3SB cells and five radioresistant mutant cell lines during the incubation after X-ray irradiation. The cells were irradiated with 5 Gy of X-rays, and nuclear extracts were prepared immediately after irradiation or 0.5 and 3 h after exposure. DNA binding activity was analyzed by an EMSA as described in “Materials and Methods.” b, Western blot analysis of the expression of NF-κB and IκBα in 3SB and 1B1C4 cells at various times after X-ray irradiation. The cells were irradiated with 5 Gy of X-rays, and cytoplasmic extracts were prepared immediately after irradiation or 0.5, 1.0, and 3.0 h after exposure.
- Fig. 4.
Apoptosis and NF-κB activity in X-irradiated mouse thymocytes. a, time course of the appearance of apoptotic cells after X-ray irradiation with 5 Gy. The percentage of apoptotic cells was calculated as the percentage of cells stained with erythrosin B in thymocytes derived from p53 wild-type (+/+) (•), heterozygous (KO/+) (○), and homozygous null (KO/KO) (□) mice. b, changes in NF-κB DNA binding activity in p53 wild-type (+/+), heterozygous (KO/+), and homozygous null (KO/KO) thymocytes during the incubation after X-ray irradiation. The freshly isolated thymocytes were irradiated with 5 Gy of X-rays, and nuclear extracts were prepared immediately after irradiation or 3, 6, and 12 h after exposure. DNA binding activity was analyzed by an EMSA as described in “Materials and Methods.”
Volume 59, Issue 24
