Fig. 1
Citrullinated myelin inhibits remyelination. A Western blot analysis of liver and myelin preparations citrullinated ex vivo by incubation with PAD2 in the presence of high calcium. B Western blot analysis of unmodified and citrullinated myelin probed for citrulline (CIT) and myelin basic protein (MBP). C Analysis of PAD2 enzyme carry-through in the modified myelin preparation. D Evidence that pHrodo red-labeled myelin is phagocytosed by microglia around the injection site following stereotactic delivery of 10 μg myelin above the corpus callosum. Scale bar is 100 μm. E Schematic showing site of intracranial myelin injection and region of analysis for MBP surface area. Representative images of MBP-labeled tracts radiating from the cingulum bundle in F control (CON) mice that were never demyelinated, G cuprizone (CUP)-fed mice at the end of the demyelination phase, H mice that received vehicle (VEH) injection, I mice that received unmodified (UNMOD) myelin, and J mice that received citrullinated (CIT) myelin by intracranial injection. K Quantitative Neurolucida analysis of MBP surface area along fibers within a 0.4 mm2 ROI. Each symbol represents one animal; CON, n = 6; CUP, n = 6; VEH, n = 24; UNMOD, n = 23; CIT, n = 26. Graph shows mean ± 95% CI. Pairwise comparisons not shown are insignificant