Forensic Science International Interleukin (IL)-8 immunoreactivity of injured axons and surrounding oligodendrocytes in traumatic head injury  Takahito Hayashi, Kazutoshi Ago, Takuma Nakamae, Eri Higo, Mamoru Ogata  Forensic Science International  Volume 263, Pages 48-54 (June 2016) DOI: 10.1016/j.forsciint.2016.03.055 Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Immunostaining for AβPP in a head injury case with 9 days survival (Case 7 in Table 1). Axonal bulbs and injured axons are labeled by AβPP. (a) Representative image of Pattern 1 showing AβPP immunoreactive axons oriented along white matter bundles. (b) Representative image of Pattern 2 showing AβPP immunoreactive axons scattered irregularly in white matter. Original magnification, 200×. Forensic Science International 2016 263, 48-54DOI: (10.1016/j.forsciint.2016.03.055) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Immunostaining for AβPP in control cases (a, Case 1; b, Case 4; c, Case 7 in Table 2) and for IL-8 in serial sections of each case (d, Case 1; e, Case 4; f, Case 7 in Table 2). In e, arrows and arrowheads indicate non-specific staining of vessel walls and amyloid bodies, respectively. The sections of immunostaining for IL-8 (d–f) were followed by nuclear staining with hematoxylin. Original magnification, 100×. Forensic Science International 2016 263, 48-54DOI: (10.1016/j.forsciint.2016.03.055) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Immunostaining for IL-8 in head injury cases (a, Case 15; b, Case 12 in Table 1). A portion of AβPP immunoreactive axonal bulbs and injured axons are labeled by IL-8. The arrows indicate IL-8 immunoreactive axonal bulbs. Original magnification, 200×. Forensic Science International 2016 263, 48-54DOI: (10.1016/j.forsciint.2016.03.055) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Immunostaining for AβPP (a) and IL-8 (b) in semi-serial sections of head injury cases showing both staining patterns of AβPP immunoreactivity (Case 7 in Table 1). The boxes with the continuous line indicate Pattern 1 and those with the dotted line indicate Pattern 2. Original magnification, 100×. Forensic Science International 2016 263, 48-54DOI: (10.1016/j.forsciint.2016.03.055) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Double immunofluorescence analysis. A sample of a head injury case (Case 7 in Table 1) was immunostained with coupled antibodies, anti-IL-8 mAb [i, Cy3] and anti-MBP pAbs [ii, FITC], and observed under a fluorescence microscope. Signals in i and ii were digitally merged in iii. White arrowheads indicate IL-8 immunoreactive oligodendrocytes. Original magnification, 200×. Forensic Science International 2016 263, 48-54DOI: (10.1016/j.forsciint.2016.03.055) Copyright © 2016 Elsevier Ireland Ltd Terms and Conditions