Prolonged increase in rat hippocampal chemokine signalling after status epilepticus  Anne A. Kan, W. Saskia van der Hel, Sharon M. Kolk, Ineke W.M. Bos, Suzanne A.M.W. Verlinde, Onno van Nieuwenhuizen, Pierre N.E. de Graan  Journal of Neuroimmunology  Volume 245, Issue 1, Pages 15-22 (April 2012) DOI: 10.1016/j.jneuroim.2012.01.012 Copyright © 2012 Elsevier B.V. Terms and Conditions

Fig. 1 Longitudinal expression patterns of IR for chemokine signalling proteins after SE. Photomicrographs of the hippocampus showing typical examples of IR of chemokines CCL2 (A) and CCL4 (B), chemokine receptors CCR1 (C) and CCR5 (D), interleukin IL-6 (E) and microglial marker Iba1 (F) at 2, 8 and 19weeks (2w, 8w and 19w) after SE and saline treated controls (2w). Note IR up-regulation is predominantly found in the hilar region (h). CCR5 and IL-6 IR up-regulation is also found in the molecular layer (ML) and the stratum radiatum (SR) 8 and 19weeks after SE. CCL4 and CCR5 IR is counterstained with haematoxylin. Scale bar=500μm. Journal of Neuroimmunology 2012 245, 15-22DOI: (10.1016/j.jneuroim.2012.01.012) Copyright © 2012 Elsevier B.V. Terms and Conditions

Fig. 2 Quantification of hilar IR for chemokine signalling proteins at various time-points after SE. Relative optical densities (ROD) of the hilar IR for CCL2 (A), CCL4 (B), CCR1 (C), CCR5 (D), IL-6 (E) and Iba1 (F) were determined at 2w, 4w, 8w and 19weeks after SE (n=6 animals per time-point). Data were normalized to time-matched controls (n=5 animals per time-point). *Significantly different P<0.05, **significantly different P<0.01. Journal of Neuroimmunology 2012 245, 15-22DOI: (10.1016/j.jneuroim.2012.01.012) Copyright © 2012 Elsevier B.V. Terms and Conditions

Fig. 3 CCL2 IR was predominantly found in neurons and CCL4 IR predominantly in (reactive) astrocytes and vascular endothelial cells. Fluorescent micrographs showing typical examples of double label immunofluorescent (IF) experiments in the hilar region of the hippocampus 2 (A, C) and 19weeks (B, D) after SE. Chemokines are in green, cell markers in red. Insets (green, red, and merge) are confocal images of representative cells in the same region. CCL2 IF (A and B) co-localized mostly with neuronal marker NeuN and was also found in a small subset of hilar Iba1-positive cells. CCL4 IF (C, D) was predominantly found in GFAP-positive astrocytes. CCL4 IF was also detected in a subset of Vimentin-positive cells; at 2w these were both activated astrocytes and endothelial cells and at 19 w only endothelial cells of the microvasculature showed IF for both. Scale bar=100μm. Journal of Neuroimmunology 2012 245, 15-22DOI: (10.1016/j.jneuroim.2012.01.012) Copyright © 2012 Elsevier B.V. Terms and Conditions

Fig. 4 CCR5 and IL-6 IR was mainly found in GFAP- and Vimentin-positive glial cells. Fluorescent micrographs showing typical examples of double label immunofluorescence (IF) experiments in the hilar region of the hippocampus 2 and 19weeks after SE. CCR5 and IL-6 are in green, cell markers in red. Insets are confocal images of representative cells in the same region. CCR5 and IL-6 IF co-localized with GFAP- and Vimentin-positive astrocytes 2 w after SE. 19 w after SE CCR5 and IL-6 IF was found in GFAP-positive astrocytes and IL-6 was also detected in the Vimentin positive endothelial cells of the microvasculature. Scale bar=100μm. Journal of Neuroimmunology 2012 245, 15-22DOI: (10.1016/j.jneuroim.2012.01.012) Copyright © 2012 Elsevier B.V. Terms and Conditions