Fibrinogen deficiency suppresses the development of early and delayed radiation enteropathy

Figure 1 PCR analysis of mouse genotype and fibrinogen levels in plasma and intestinal tissue of Fib^+/+, Fib^+/- and Fib^-/- mice. A: Representative agarose gel image showing DNA marker (Lane 1), single band at around 376 base pair (Lane 2 and 3) that denotes Fib^+/+ mice, double bands, one at around 376 bp and the other is at around 283 bp (Lane 4 and 5), which signify Fib^+/- mice, and single band at around 283 bp (Lane 6 and 7) that represents Fib^-/- mice; B: Plasma fibrinogen level in un-irradiated Fib^+/+ (n = 5), Fib^+/- (n = 6) and Fib^-/- (n = 5) mice as detected by enzyme-linked immunosorbent assay (ELISA) assay; C: Fibrinogen level in the intestinal tissue of un-irradiated Fib^+/+ (n = 6), Fib^+/- (n = 10) and Fib^-/- (n = 3) mice as detected by ELISA assay.

Figure 2 Fibrinogen immunoreactivity in the intestinal tissue of Fib^+/+, Fib^+/- and Fib^-/- mice before and after irradiation. A: Representative photomicrograph showing fibrinogen level in un-irradiated Fib^+/+ mice; B: Fibrinogen level in un-irradiated Fib^+/- mice; C: Fibrinogen level in un-irradiated Fib^-/- mice; D: Fibrinogen level in irradiated Fib^+/+ mice; E: Fibrinogen level in irradiated Fib^+/- mice; F: Fibrinogen level in irradiated Fib^-/- mice; and G: Column diagram showing comparison of intestinal fibrinogen levels among un-irradiated (n = 6) and irradiated (n = 6) Fib^+/+ mice, un-irradiated (n = 10) and irradiated (n = 10) Fib^+/- mice, and un-irradiated (n = 3) and irradiated (n = 3) Fib^-/- mice.

Figure 3 Morphological changes in the intestinal tissue of Fib^+/- and Fib^-/- mice observed at 2 wk and 26 wk after localized irradiation. A: Comparative assessment of structural radiation injury between Fib^+/- (n = 15) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 28) and Fib^-/- (n = 11) mice at 26 wk; B: Comparative assessment of intestinal wall thickness between Fib^+/- (n = 14) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 24) and Fib^-/- (n = 11) mice at 26 wk; C: Comparative assessment of intestinal serosal thickness between Fib^+/- (n = 14) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 24) and Fib^-/- (n = 11) mice at 26 wk.

Figure 4 Intestinal inflammation and intestinal mucosal injury in Fib^+/- and Fib^-/- mice observed at 2 wk and 26 wk after localized irradiation. A: Comparative assessment of intestinal inflammation between Fib^+/- (n = 13) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 28) and Fib^-/- (n = 13) mice at 26 wk by measuring myeloperoxidase activity; B and C: Representative photomicrographs showing myeloperoxidase positive cells in Fib^+/- and Fib^-/-, respectively, at 2 wk after exposure; D: Comparative assessment of intestinal mucosal injury between Fib^+/- (n = 15) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 28) and Fib^-/- (n = 11) mice at 26 wk; E and F: Representative photomicrographs showing difference in mucosal surface area by H&E staining in Fib^+/- and Fib^-/-, respectively, at 2 wk after exposure.

Figure 5 Intestinal smooth muscle cell proliferation, collagen deposition and TGF-β immunoreactivity in Fib^+/- and Fib^-/- mice observed at 2 wk and 26 wk after localized irradiation. A: Comparative assessment of intestinal smooth muscle cell proliferation between Fib^+/- (n = 13) and Fib^-/- (n = 13) mice at 2 wk and Fib^+/- (n = 19) and Fib^-/- (n = 12) mice at 26 wk by measuring PCNA immunoreactivity; B and C: Representative photomicrographs showing PCNA immunoreactivity in Fib^+/- and Fib^-/-, respectively, at 2 wk after exposure; D: Comparative assessment of collagen deposition between Fib^+/- (n = 11) and Fib^-/- (n = 14) mice at 2 wk and Fib^+/- (n = 17) and Fib^-/- (n = 10) mice at 26 wk; E and F: Representative photomicrographs showing Collagen deposition by Trichrome staining in Fib^+/- and Fib^-/-, respectively, at 2 wk after exposure; G: Comparative assessment of TGF-β immunoreactivity between Fib^+/- (n = 13) and Fib^-/- (n = 13) mice at 2 wk and Fib^+/- (n = 21) and Fib^-/- (n = 12) mice at 26 wk; H and I: Representative photomicrographs showing TGF-β immunoreactivity in Fib^+/- and Fib^-/-, respectively, at 2 wk after exposure.