Journal: International Journal of Molecular Sciences

Article Title: Systemic Lipopolysaccharide Challenge Induces Inflammatory Changes in Rat Dorsal Root Ganglia: An Ex Vivo Study

doi: 10.3390/ijms232113124

Figure Lengend Snippet: Nuclear translocation of inflammatory transcription factors nuclear factor interleukin 6 (NF-IL6) and signal transducer and activator of transcription 3 (STAT3) in cultured dorsal root ganglia (DRG) cells. Immunocytochemistry was performed in cultured DRG cells obtained from PBS- and LPS-treated rats to examine activation of inflammatory transcription factors. ( A ) In CD-68-positive macrophages (green) an enhanced nuclear signal of NF-IL6 (red) was detectable in rats treated with LPS ( A.2 ) compared to controls ( A.1 ). Calculating the mean nuclear intensity of the signal in the area of the nucleus (blue), a significant increase is detectable ( A.3 ). ( B ) The STAT3 signal (red) was mainly detectable in MAP-positive neurons (green) of DRG cultures of LPS-treated animals ( B.2 ) and controls ( B.1 ). Calculating the mean nuclear intensity of the signal in the area of neuronal nuclei (blue), a significant difference is detectable ( B.3 ). Bars represent the mean ± SEM. ‘ n ’ represents the number of investigated cells of the respective cell type. Scale bars present 10 µm. **: p < 0.01; ****: p < 0.0001. PBS: phosphate buffered saline, LPS: lipopolysaccharide.

Article Snippet: For immunocytochemical identification of cell types, the following monoclonal antibodies or polyclonal antisera were used: CD-68 for macrophages (mouse anti rat-CD-68; 1:1000; AbD Serotec, Oxford, UK) and microtubule-associated protein 2a+b for neurons (mouse AP-20 anti-MAP2a+b; 1:600; Sigma-Aldrich Chemie GmbH).

Techniques: Translocation Assay, Cell Culture, Immunocytochemistry, Activation Assay, Saline

Journal: Journal of neuroscience methods

Article Title: A novel technique for morphometric quantification of subarachnoid hemorrhage-induced microglia activation

doi: 10.1016/j.jneumeth.2014.04.001

Figure Lengend Snippet: No change in microglial position with respect to the nearest capillary, or in expression of the histochemical activation marker CD-68 could be detected. A. Microglial position with respect to the nearest capillary, regardless of whether the cell is activated or not, did not change 1, 2, or 7 days following SAH (n = 3–9 animals per group). B., C. The raw count of microglia, as well as the subpopulation which are eGFP+/CD-68+ and thus activated, did not change 1, 2, or 7 days following SAH (n = 5–13 animals per group). D. The proportion of all microglia that are activated also did not change 1, 2, or 7 days following SAH (n = 5–13 animals per group). Key: green signal = eGFP, red signal = CD-31, magenta signal = CD-68, and blue signal = DAPI. All bars graphs represent mean +/− SEM.

Article Snippet: 2.4 Immunohistochemistry: Staining Protocol The primary antibodies used were both diluted to 1:200 and included rat anti-mouse CD-68 (macrosialin, a marker of reactive macrophagic microglial cells) (MCA1957, Serotec) and rabbit anti-mouse CD-31 (PECAM-1, a marker of vascular endothelium) (ab28364, Abcam).

Techniques: Expressing, Activation Assay, Marker