Figure 1.
Nucleated cells and stem/progenitor cells in the peripheral blood of mice after treatment.
The number of nucleated cells in the peripheral blood (A) was directly counted, and c-kit+ (B) and CD34+ stem/progenitor cells (C) in the peripheral blood were measured in the fraction of the nucleated cells by flow cytometry. The open circles represent data from each mouse and the red lines indicate median values of each group.
Figure 2.
Stem/progenitor cells in the bone marrow of mice after the treatments.
Bone marrow mononuclear cells were collected 2 days after the last radiation treatment, and the c-kit+ (A) and CD34+ (B) stem/progenitor cells were measured by flow cytometry. The open circles represent data from each mouse and the red lines indicate median values of each group.
Figure 3.
Bone marrow mononuclear cells were collected 2 days after the last radiation treatment. Freshly isolated bone marrow mononuclear cells were mixed in methylcellulose complete medium, and the colony formation was observed under microscopy at 9 days after incubation. A) Different types of colonies, including CFU-GEMM, CFU-M, CFU-GM, and BFU-E were clearly formed from the bone marrow cells. B) A significantly higher number of total colonies (>50 cells) was formed from the bone marrow cells of the mice that were given nicaraven than those that received placebo. The open circles represent the mean of data from a mouse with duplicate assay. The red lines indicate the median values of each group.
Figure 4.
DNA damage in bone marrow cells.
Bone marrow cells from mice were seeded in 4-well culture slides and cultured for 7 days. The DNA damage in the cells was estimated by immunostaining with an anti-53BP1 antibody. A) Representative images show the formation of 53BP1 foci within the nuclei of some cells (arrowheads). B) Quantitative analysis shows that the percentages of cells with 53BP1 foci were significantly lower in the Nicaraven group than the Placebo group. The open circles represent data from each mouse and the red lines indicate median values of each group.
Figure 5.
Intracellular and mitochondrial ROS in bone marrow cells.
Mice were exposed daily to 1 Gy γ-rays for 5 days in succession, and either nicaraven or a placebo was given after each exposure. The cells were collected 2 days after the last radiation treatment and then loaded with 10 µM CM-H2DCFDA or 5 µM MitoSOX Red at 37°C for 30 minutes. The intracellular ROS (A) and mitochondrial ROS (B) were detected as the mean fluorescence intensity in all cells by flow cytometry. The open circles represent data from each mouse and the red lines indicate median values of each group.
Figure 6.
The levels of 8-OHdG in the plasma and urine.
The plasma 8-OHdG levels were very low and did not significantly differ between the two groups (A), but the levels of urinary 8-OHdG were significantly lower in the mice that received nicaraven than in those that received a placebo (B). The open circles represent the mean of data from a mouse with duplicate assay. The red lines indicate median values of each group.
Figure 7.
Inflammatory cytokines in the plasma.
The plasma was collected 2 days after the last radiation treatment, and the levels of the inflammatory cytokines IL-6 (A) and TNF-α (B) in the plasma were measured by ELISA. The open circles represent the mean of data from a mouse with duplicate assay. The red lines indicate median values of each group.