xoplasma and Neospora tachyzoites using Trizol. Nuclease digestions were carried out using 60 mg of total RNA in 15 ml of volume containing 3 units/ml of S1 nuclease, alone or following Dnase treatment and RNA PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19645759 purification, and incubated at 37uC for 45 minutes. Samples were analyzed on 0.8% native agarose in 16 TAE buffer at 4uC. S1 nuclease-treated Leishmania guyanensis, which contains the endogenous RNA virus LRV1, was used as a positive control. Fluorescence microscopy to assess parasite internalization HFF cells grown to confluency on coverslips were infected with a 3:1 MOI of either live or heat-killed RH strain Toxoplasma expressing mCherry. 18 hours after infection, coverslips incubated with 50 nM LysoTracker for 10 minutes at 37uC were live mounted, and immediately imaged using a Leica DMI400 with Yokagawa spinning disk confocal system. Images were overlayed and analyzed using Image J software, v1.47. Statistical Analysis and data visualization All experiments were repeated 2-4 times, and means and standard deviations were calculated from biological replicates. Significance was determined using a Student’s t-test. Statistical analysis was carried out using Prism 4 and data visualization using DataGraph 3.0. After injury, the central nervous system of adult mammals is limited in its ability to recover because of the inability of damaged axons to reconnect and regain their physiological 
structure and function. Factors that influence axon regeneration include neural cell-autonomous activity, glial scarring, local inflammation, and inhibition factors. In the past two decades, several CNS myelin-derived axon growth inhibitory factors have been found, including NogoA, myelin-associated glycoprotein, and oligodendrocyte myelin glycoprotein . These proteins have been the subject of great research interest and are highly clinical relevant. NogoA plays an important role in recovery from spinal cord injury, oligodendrocyte differentiation and myelination, and the development of the CNS. A recent study reported that a region of NogoA attenuates cerebral ischaemia by inhibiting NADPH oxidasemediated oxidative damage and neuronal apoptosis, indicating that the functional diversity of different fragments of NogoA must be explored. The function of NogoA has been studied 92-61-5 extensively using antiNogoA antibodies. In a previous study, we developed two anti-NogoA monoclonal antibodies, aNogoA-N and aNogo66 mAb, which were generated in mice using recombinant aa 570 691 and aa 10261091 fragments, respectively, from NogoA. In the present study, we analysed the specificity and affinity of the two mAbs to the NogoA molecule. We also detected the different epitopes in NogoA that could be recognised by the two mAbs. Using in vitro experiments, we found that these mAbs against NogoA enhanced axon growth and branch formation. Materials and Methods Animals Male SpragueDawley rats weighing 200220 g and Sprague Dawley rat embryos were obtained from the Experimental Animal Center of the Fourth Military Medical University. All experimental procedures were approved by the Ethics Committee for Animal Experimentation of the Fourth Military Medical University. The protocols used in this research project complied with the guidelines for the care and use of laboratory animals of the Fourth Military Medical University. During the experiments, all efforts were made to minimise animal suffering and the number of animals used. 1 Antibodies of NogoA Enhance Axon Extension A