Figure 5

Visualization of immune mediators and their producer cells in the CNS of B. burgdorferi -infected animals. A) Indication of the cytokine IL-6 (green) due to staining with anti-IL-6 antibody followed by secondary antibody conjugated with Alexa fluor 488, in astrocytes (red), due to staining with anti-GFAP-cy3 in the spinal cord of animal DH50. The yellow signal is due to co-localization of the astrocytic marker GFAP (red) and IL-6 (green) within the astrocytes. B) The cellular localization of IL-6 (green) in neurons appearing red due to staining with anti-NeuN antibody, followed by secondary antibody conjugated with Alexa fluor 568. As NeuN is a nuclear antigen it is seen to stain the neuronal nucleus, while IL-6 is evident in the cytoplasm and extracellularly. C) The presence of CXCL13 (green) due to staining with anti-CXCL13 antibody followed by secondary antibody conjugated with Alexa fluor 488, in microglia (red) due to staining with anti-IBA-1 followed by secondary antibody conjugated to Alexa fluor 568 in the spinal cord of animal DH50. Intracytoplasmic staining of CXCL13 in microglia appears yellow. D) Evidence of CCL2 (green) due to staining with anti-CCL2 antibody followed by secondary antibody conjugated with Alexa fluor 488, in microglia (red) in the spinal cord of DH50. E). A representative image of the absence of IL-6 (which should have appeared green due to labeling with anti-IL-6 antibody followed by secondary antibody conjugated to Alexa fluor 488) in astrocytes labeled with anti-GFAP-cy3 (red) in the spinal cord of a control animal. F). Absence of IL-6 in neurons appearing red due to staining with anti-NeuN antibody, followed by secondary antibody conjugated with Alexa fluor 568 in the DRG of a control animal. G). Absence of CCL2 in microglia (red) due to staining with anti-IBA-1 followed by secondary antibody conjugated to Alexa fluor 568 in the spinal cord of control animals. The unstained tissue in all images appears gray due to differential interference contrast (DIC) imaging.