Sera N, Hida A, Imaizumi M, Nakashima E, Akahoshi M. were not observed at earlier time points. These results suggest that late effects of TBI include medial growth due to collagen deposition. All images were acquired at 400; level pub = 50 m. Fig. S2. Representative images (400) of H&E stained remaining coronary artery mix sections from TBI and nonirradiated age-matched control mice at 4 and 18 months post- TBI. Arrows show the location of intimal nuclei, which were reduced in quantity in the TBI mice whatsoever ages. There was no apparent decrease in the number of medial cell nuclei. The intimal cell nuclei were all consistent with an endothelial phenotype. Some of the medial cells were binucleate (observe asterisk), an indication of senescence. Fig. S3. Representative images (400) of -actin-positive arterioles in TBI and age-matched nonirradiated myocardium at 13 weeks post-TBI. Images were acquired from formalin-fixed paraffin-embedded sections of heart reacted with an antibody to -clean muscle mass cell actin, as explained in Bavisant Materials and Methods, and used to determine arteriolar denseness. Arteriole figures per unit area in the remaining ventricle of TBI mice decreased significantly at 13 weeks compared to nonirradiated controls, and related results were obtained at 18 months post-TBI. Fig. S4. Representative images of remaining ventricle and coronary artery mix sections Bavisant stained with Perls Prussian blue from TBI and age-matched nonirradiated control mice at 4 weeks post-TBI, and additional TBI cells at 6 and 18 months. All images were acquired at 400. Perls-positive areas (hemosiderin deposits) were observed in the myocardium and coronary arteries of TBI mice. Sites of deposition included the myocardium and epicardium and, in arteries, the peri-arterial space, medial layer and intima. Fig. S5. Representative images of the renal cortex stained with PSR in TBI and age-matched nonirradiated control mice at 6, 13 and 18 months postirradiation. All images were acquired at 400; level pub = 50 m (demonstrated in the panel labeled 18 months nonirradiated) applies to all images. ITGAL Improved PSR staining in the renal cortex was observed 1st in the glomeruli of TBI mice and later on (18 months) in the interstitium, especially in regions of tubular atrophy. Fig. S6. Representative images of PSR (top 4 panels; 400) and H&E stained mix sections (bottom 2 panels) of renal arteries from TBI and age-matched nonirradiated control mice. Fewer renal arteries were available for analysis than coronary arteries as not all kidney sections were obtained at the location which contained these arteries. In the PSR-stained sections, some renal arteries from TBI mice appeared to have more collagen in the adventitia and press at later on times post-TBI; however, there was significant variance within and between mice of both the nonirradiated and TBI organizations. As demonstrated in the H&E stained sections (bottom 2 panels), no stunning variations were observed in intimal cell nuclear quantity for either large arteries or arterioles, contrary to what was observed in the heart. Fig. S7. Representative images (400) of -actin positive arterioles in TBI and age-matched nonirradiated renal cortex at 18 months. Images were acquired from formalin-fixed paraffin inlayed sections of Bavisant kidney reacted with an antibody to -clean muscle mass cell actin as explained in Materials and Methods and used to determine arteriolar denseness. A decrease in arteriole quantity per unit area was significant only at 18 months post-TBI. Fig. S8. Representative images of kidney mix sections stained with Perls Prussian blue from TBI and age-matched nonirradiated control mice. Perls staining was significantly elevated in the renal cortex of most irradiated mice at 4 weeks post-TBI, was reduced to near-nonirradiated ideals in most TBI mice at 6 months (3 of 4) and then became elevated in 50% of the mice (5 of 10) at 13C18 weeks post-TBI. Bavisant Perls staining was observed primarily in the tubules of the superficial cortex region, but not all tubules within a region exhibited staining. Staining was also observed in some glomeruli at later on time points. Perls positive staining or deposits of hemosiderin were not observed in the renal arteries, even when the arteries were surrounded by tubules with hemosiderin. Fig. S9. Representative images (400) of macrophages in mix sections of heart and kidney reacted with the F4/80 antibody. Panel A: Age-matched nonirradiated mouse heart remaining ventricle experienced an extremely low macrophage denseness. Panel B: Four-months post-TBI, mouse heart left ventricle shown a significantly improved quantity of macrophages (arrows) compared to nonirradiated controls. Panel C: Renal macrophages were essentially undetectable at any time point in nonirradiated kidney sections and only occasionally in.