SOD1G93A mouse ventral horn cervical spinal cord tissue immuinostained for ephrinB2 at 60 days (b), 120 days (c), endstage (d), and WT mouse age-matched control (a), scale bar: 200 µm. …
File contains the raw data for Figure 1, panel E.
We injected 60-day-old SOD1G93A mice with Lenti-GFP control or Lenti-shRNA-Efnb2 into cervical ventral horn (a). Injections were made bilaterally into six sites throughout C3-C5 to target the PhMN …
File contains the raw data for Figure 2, panels M and N.
SOD1G93A mice injected with Lenti-GFP control or Lenti-shRNA-Efnb2 were cresyl violet stained and MN counts were performed at 117 days of age. Thirty µm transverse cervical spinal cord tissue …
File contains the raw data for Figure 3, panels B and G.
Biweekly weights of Lenti-shRNA-Efnb2 and Lenti-Control mice were recorded until endpoint sacrifice (a), and disease onset was measured for each animal when there was a 10% drop in body weight (b). …
File contains the raw data for Figure 4, panels A-K.
Diaphragm muscles were labeled with SMI-312 (green), SV2, (green) and alpha-Bungarotoxin (blue). Representative images of fully-innervated (a), partially-denervated (b) and completely-denervated ((c)…
File contains the raw data for Figure 5, panels F-H.
Immunoblotting analysis on postmortem samples from human ALS donors with an SOD1 mutation and also non-diseased human samples. In the lumbar enlargement, there was a large increase in ephrinB2 …
File contains the raw data for Figure 6.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (Mus musculus) | Transgenic SOD1G93A mice: C57BL/6 J congenic line B6.Cg-Tg(SOD1*G93A)1Gur/J | The Jackson Laboratory | RRID: IMSR_JAX:004435 | Both female and male mice used |
Genetic reagent (Mus musculus) | Transgenic SOD1G93A mice: B6SJL-Tg(SOD1*G93A)1Gur/J | The Jackson Laboratory | RRID:IMSR_JAX:002726 | Both female and male mice used |
Biological sample (Homo sapiens) | Spinal cord and cortex samples from ALS donors | Biorepository of the Jefferson Weinberg ALS Center (1 of 3 ALS samples) Project ALS (2 of 3 ALS samples) | All three ALS donors had an SOD1 mutation (donor 1: D102H mutation; donor 2: A4V; donor 2: V87A), and all three donors did not have a C9orf72 repeat expansion. These three donors succumbed to ALS at 42 (female), 55 (male), or 58 (male) years of age. | |
Biological sample (Homo sapiens) | Spinal cord and cortex from non-ALS donors | NIH NeuroBioBank | Age of death for these three non-ALS donors was 67, 70, and 70 years. | |
Antibody | Mouse monoclonal Anti-SV2 (Used for IHC) | Developmental Studies Hybridoma Bank, Iowa City, IA | RRID: AB_2315387 | 1:10 |
Antibody | Mouse monoclonal Anti-SMI312 (Used for IHC) | Covance, Greenfield, IN | RRID: AB_2314906 | 1:1000 |
Antibody | Mouse monoclonal Anti-NeuN (Used for IHC) | EMD-Millipore, Temecula, CA | RRID: AB_2298772 | 1:200 |
Antibody | Rabbit polyclonal Anti-GFAP (Used for IHC) | Dako, Carpinteria, CA | RRID: AB_10013482 | 1:400 |
Antibody | Mouse monoclonal Anti-GFAP (Used for Westerns) | BD Bioscience, Franklin Lakes, NJ | Cat. #610566 | 1:2000 |
Antibody | Rabbit polyclonal Anti-Olig2 (Used for IHC) | EMD-Millipore, Temecula, CA | RRID: AB_2299035 | 1:200 |
Antibody | Goat polyclonal Anti-ephrinB2 (Used for IHC) | R&D Systems, Minneapolis, MN | RRID: AB_2261967 | 1:50 |
Antibody | Rabbit polyclonal Anti-ephrinB2 antibody (Used for Westerns) | Abcam, Cambridge, MA | RRID: AB_11156896 | 1:500 |
Antibody | Goat polyclonal Anti-EphA4 (Used for IHC) | R&D Systems, Minneapolis, MN | RRID: AB_2099371 | 1:100 |
Antibody | Rabbit polyclonal Anti-GFP (Used for IHC) | Aves Labs, Davis, CA | RRID: AB_10000240 | 1:500 |
Recombinant DNA reagent | Lenti-shRNA-Efnb2; VSVG.HIV.SIN.cPPT.U6. SbRmEphrinB2.4.CMV.EGFP | This study | Vector generated by the Dalva lab | |
Recombinant DNA reagent | Lenti-Control; VSVG.HIV.SIN.cPPT. U6.Empty.CMV.EGFP | This study | Vector generated by the Dalva lab | |
Software | Scope 3.5.6 software | ADInstruments, Colorado Springs, CO | RRID: SCR_001620 | |
Software | ImageJ/Fiji software | RRID: SCR_003070 | ||
Software | Bio-Rad Image Lab software | Bio-Rad, Hercules, CA | RRID:SCR_014210 | |
Software | Graphpad Prism 6 | Graphpad Software Inc, LaJolla, CA | RRID: SCR_002798 | |
Other (Microscope) | Olympus FV1000 confocal microscope | Olympus, Center Valley, PA | RRID: SCR_014215 | ‘NMJ analysis’ subsection of the Materials and methods section |
Other (Microscope) | Zeiss Axio M2 Imager confocal microscope | Zeiss, Hebron, KY | RRID:SCR_020922 | ‘Motor neuron counts’ subsection of the Materials and methods section |