Fig 2: Fluorescence images of samples of the N&A-M group and the N group on days 1, 3, 7, and 14 (the neuronal dendrites were stained with MAP2 and are shown in red; the astrocytes were stained with GFAP and are shown in green; the cell nuclei were stained with DAPI and are shown in blue)

Fig 3: DRG neurons promote the migration and invasion of EC cells. (A) EC tissue with or without PNI was stained in Hematoxylin and eosin (HE). The nerve area was marked by dashed lines. N = nerves; C = cancer cells. (B-C) DRG was placed in the center of Matrigel. The interaction of EC cell line and DRG was confirmed in bright field (black arrow) and immunofluorescence staining (yellow arrow). ISK cells were stained by pan-cytokeratin (pan-CK), green, scale bar, 100 µm. (D) Representative bright field image of DRG neurons after culture for 3 days (black arrow); scale bar, 50µm. (E) Immunofluorescence staining of DRG neurons. The dissociated DRG neurons were defined as DAPI+/MAP2+ (red arrow: DRG neuron, yellow arrow: nerve fiber); scale bar, 50µm. (F) Transwell coculture systems were involved to evaluate the migration/invasion effect of DRG neurons on EC cells; scale bar, 100µm. (G) The migrated/invaded cells were then counted in five random fields and statistically analyzed. Data are presented as the mean±SD; **P<0.01.

Fig 4: Identification of the autoantibody in a patient with acute encephalitis. P1 patient was diagnosed with encephalitis. The CSF sample from a control patient with carpal tunnel syndrome (nCSF) and the serum from a healthy person (nSerum) was used as negative controls. (A) Immunostaining on rat brain sections. The CSF from P1 patient had stained on hippocampus and cerebellum. (B) P1 patient’s serum stained obviously in the cytosolic compartments of the neurons in the cortex, CA3 area of the hippocampus, and cerebellum on mouse brain sections (green). White arrows, possible oligodendrocytes; arrow heads, Purkinje cells. CRMP2-expressing HEK293T cell absorbed P1 patient’s serum had no obvious staining. Cell nuclei were stained blue with DAPI. (C) P1 patient’s serum (green) stained positively in the soma and dendrites of cultured mouse cortical neurons which were labeled with anti-MAP2 Ab (red). (D, E) IP of P1 CSF with rat brain protein lysate. In total, 500 µl (lane 1) and 100 µl (lane 2) P1 CSF were applied to IP with 10 mg total protein of fresh rat brain lysate, respectively. Moreover, 500 µl nCSF was used for negative control. A positive band around 70 kDa was pulled down by P1 CSF but not by nCSF. A similar protein size band (red arrowhead) was observed on a parallel gel with Coomassie brilliant blue staining (E). (F) Characterization of the autoantigen by mass spectrometry. The suspected protein band in (E) was collected for protein identification by LC-MS/MS. Seventeen identified peptides (red) matched rat DPYL2 (UniProtKB ID: P47942), an alias of CRMP2, with a score of 226 and sequence coverage of 40%. CRMP2, collapsin response mediator protein 2; CSF, cerebrospinal fluid; DAPI, 6-diamidino-2-phenylindole; DPYL2, dihydropyrimidinase like 2; GL, granular layer; IB, immunoblotting; IHC, immunohistochemistry; IP, immunoprecipitation; LC-MS/MS, liquid chromatography tandem mass spectrometry; MAP2, microtubule-associated protein 2; ML, molecular layer; PCL, Purkinje cell layer; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum. Scale bars represent 50 µm in (B) and 20 µm in (C).

Fig 5: PBM promoted motor function recovery and alleviated mitochondrial-related neuronal apoptosis. (A): Representative gaits of each group at 7, 14, and 28 dpi. The stride length was calculated, n = 3. (B): TUNEL (red) and NeuN (green) co-stained and the arrows point to representative cells. The ratio of TUNLE-positive neurons was calculated, Scale bar: 50 µm, n = 3. (C): Neurons was stained with MAP2, and nuclei were stained with DAPI (blue), Scale bar: 50 µm. (D): Western blot analysis and quantification of Bax/Bcl2 levels in each group, n = 4.