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Rat anti MBP antibody, clone 12 (MCA409S) used for the detection of myelin basic protein by immunofluorescence.
P0 rescues myelination, Caspr, and neurofascins in plp-dKO DRG cultures. Confocal coimmunofluorescence images of (A) MBP (red) and neurofilament (NF, green), or (C) P0 (red) and MBP (green), or (D) Caspr (red), total neurofascins (NFasc, green), and MBP (white), or (F) MBP (red), NF (white), and either GFP, P0, or Myc (green), or (H) Caspr (red) and NFasc186 (false-colored green) in myelinated control and plp-dKO DRG cultures with (A,C,D,F,H) or without (A) tamoxifen. Briefly, A–D demonstrate demyelination, loss of P0 and paranodal/nodal defects in plp-dKO DRG cultures, mimicking the in vivo phenotype of dKO sciatic nerves. In (F,H), plp-dKO DRG cultures were transduced with doxycycline-inducible lentiviruses expressing either GFP, P0, or P0-myc. (B,G) Quantification of MBP fluorescence intensity normalized to NF. (E,I) Percentage of intact nodes/paranodes expressing Caspr and high levels of NFasc. F–I show that exogenously delivered P0 significantly rescues demyelination and paranodal/ nodal defects of plp-dKO DRG cultures. In (C), white arrows indicate MBP positive/P0 negative fibers, and magenta arrowheads MBP/P0 double positive fibers. Merges MBP/P0 (C) or Caspr/NFasc (D) appear yellow. In (D), dashed lines delineate the paranodal region. In (H), control and plp-dKO were transduced with lentiviruses expressing GFP, and plp-dKO + P0 with lentiviruses expressing P0 (white). Z-series projections (A,C,F) and single optical sections (D,H) are shown. In (D,H), arrows indicate heminodes or full nodes. Images on the right are magnifications of white boxes depicted on the left images. At least three control and three plp-dKO embryos were used for quantification (average of three coverslips per embryos), representative images are shown. Nuclei are labeled in blue with DAPI. P-values (unpaired [B,E,I] or paired [G] two-tailed Student's t test, unless stated otherwise in the figure): * = p < 0.05, *** = p < 0.001, error bars = SEM.
From: Brügger V, Engler S, Pereira JA, Ruff S, Horn M, Welzl H, et al. (2015) HDAC1/2-Dependent P0 Expression Maintains Paranodal and Nodal Integrity Independently of Myelin Stability through Interactions with Neurofascins.
PLoS Biol 13(9): e1002258.
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