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. Author manuscript; available in PMC: 2009 Sep 18.
Published in final edited form as: J Comp Neurol. 2009 Jan 10;512(2):243–255. doi: 10.1002/cne.21902

Figure 2. ADAM8 mRNA and protein is increased following SCI.

Figure 2

A) Western blots show that of whole spinal cord extract from uninjured (0) and 7 day injured (7d SCI) mice probed with ADAM8 antibody shows two bands around the expected 80 kDa size of mature ADAM8. The doublet probably represents different post-translationally modified forms. The lack of other bands shows the specificity of the antibody. A blot of trachea extract is shown as a positive control. Note the increase of ADAM8 7 days after injury compared to uninjured control, which was seen in 3 different experiments. B) End-point RT-PCR shows the absence of ADAM8 mRNA in 2 uninjured spinal cord extracts (0) and the increased expression in individual mice at both 1 and 3 days post-injury. The bands are at 459 bp, the expected amplicon size for the primer set used. For the negative control, reverse transcriptase was replaced by water. C) RT-PCR of whole spinal cord at 3 days post-injury (3d SCI whole; 2 lanes = 2 mice) was compared to that of FACS-purified spinal microvascular ECs (smvECs) pooled from 12 mice per lane 3 days following SCI. The household gene cyclophilin A (cA) is shown for comparison. Despite the much reduced mRNA levels in the isolated preparation, judged by the reduced cA mRNA, the ADAM8 mRNA remains detectable, suggesting an enrichment of ADAM8 in blood vessels. Solid arrowhead indicates the expected amplicon size of ADAM8 and the open arrowhead the expected amplicon size of cA.