Journal: Annals of neurology

Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

doi: 10.1002/ana.23756

Figure Lengend Snippet: In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).

Article Snippet: NIHMS407111-supplement-Supp_Figure_S1.tif (21M) Supp Figure S2: Supplementary Figure 2: Analysis of patients’ antibodies using a cell-based assay expressing Kv4.2 HEK 293 cells expressing Kv4.2 immunostained with patients’ serum (A, D, G, J) and a rabbit polyclonal antibody (Alomone labs, #APC-023) against Kv4.2 (B, E, H, K).

Techniques: Immunoprecipitation, Staining, Mass Spectrometry, Western Blot

Journal: Annals of neurology

Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

doi: 10.1002/ana.23756

Figure Lengend Snippet: Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.

Article Snippet: NIHMS407111-supplement-Supp_Figure_S1.tif (21M) Supp Figure S2: Supplementary Figure 2: Analysis of patients’ antibodies using a cell-based assay expressing Kv4.2 HEK 293 cells expressing Kv4.2 immunostained with patients’ serum (A, D, G, J) and a rabbit polyclonal antibody (Alomone labs, #APC-023) against Kv4.2 (B, E, H, K).

Techniques: Labeling, Marker, Immunostaining

Journal: Annals of neurology

Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

doi: 10.1002/ana.23756

Figure Lengend Snippet: The reactivity of patients’ serum with the hippocampus of wild type mice is shown in A, C, E and G. The reactivity of a rabbit polyclonal DPPX antibody with the hippocampus of wild-type mice is shown in I. Panels on the right side show the results of a similar experiment but using the hippocampus of DPPX-null mice. Note that the reactivities of the sera of the first three patients (cases 1, 2, and 3 of Table 1) and the rabbit polyclonal DPPX antibody are abrogated in the hippocampus of DPPX-null mice (panels B, D, F, J). Patient 4, not included in the table (panels G and H) showed remaining reactivity with the hippocampus of DPPX-null mice indicating that this patient had two antibodies, one against DPPX and the other against an unknown antigen. Bar = 200 μm.

Article Snippet: NIHMS407111-supplement-Supp_Figure_S1.tif (21M) Supp Figure S2: Supplementary Figure 2: Analysis of patients’ antibodies using a cell-based assay expressing Kv4.2 HEK 293 cells expressing Kv4.2 immunostained with patients’ serum (A, D, G, J) and a rabbit polyclonal antibody (Alomone labs, #APC-023) against Kv4.2 (B, E, H, K).

Techniques: