Fig 2: Effect of 5-aza treatment on CRMP2 (A), pCRMP2 (B), and a-tubulin (a-tubulin: C; Tyr-tubulin: D; Acet-tubulin: E) isoform expression. *P < 0.05.

Fig 3: The flow chart of patients selected for anti-CRMP2 antibody screening. CSF and/or serum samples from 46 patients with neurological disorders that had positive immunostaining in the neuro-cytoplasm of rat brain sections were enrolled for retrospective anti-CRMP2 antibody screening. Twelve serum samples from healthy people were used as negative controls. A 42-year-old female patient with encephalomyelitis, right dot panel P2, was tested anti-CRMP2 antibody-positive in serum. AE, autoimmune encephalitis; CBA, cell-based assay; CRMP2, collapsin response mediator protein 2; CSF, cerebrospinal fluid; TBA, tissue-based assay.

Fig 4: Verification of anti-CRMP2 antibody. (A) IP with P1 patient’s serum was performed with rat brain protein lysate. Immunoblottings were done with P1 patient’s serum (upper panel) and commercial anti-CRMP2 antibody (lower panel). (B) IP with commercial anti-CRMP2 antibody was performed with HEK293T cells overexpressing full-length human CRMP2 (isoform 1). Anti-CRMP2 antibody, P1 patient, or negative control sera were used for Western blotting, separately. (C) CRMP2-expressing HEK293T cells were immunostained with the patients’ sera and anti-CRMP2 antibodies. The sera from the first collection of P1 and P2 patients showed positive staining and co-localized with CRMP2. Serum collected from P2 patient 1.5 years after recovery showed negative staining. (D) Colocalization of P1 patient’s serum and anti-CRMP2 Ab immunostaining in cultured mouse cortical neurons. CRMP2, collapsin response mediator protein 2; DAPI, 6-diamidino-2-phenylindole; IB, immunoblotting; nIgG, normal immunoglobulin G; IP, immunoprecipitation; WB, Western blotting. Scale bars represent 10 µm in (C) and 20 µm in (D).

Fig 5: Hippocampal protein levels of neuroinflammation and neuroplasticity biomarkers in mice. Western blot analysis of (A) NLRP3, (B) caspase-1, (C) IL-18, (D) IL-1ß, (E) CRMP2, and (F) a-tubulin protein expression. (G) Densitometry analyses of the bands. Each value represents the mean ± SEM. n = 4 animals per group. Two groups (NC, IMQ) were analyzed using t-test (* p < 0.05, ** p < 0.01).