(A) C2 myotubes were treated with neural Agrin or antibodies for the indicated times. MuSK was immunoprecipitated, and Western blots were probed for MuSK or phosphotyrosine. Neural Agrin and MuSK …
(A) The MuSK agonist antibody (Ab #13) failed to stimulate MuSK phosphorylation in innervated (Inn) muscle, suggesting that MuSK may be maximally phosphorylated at synapses by Agrin and Lrp4. (B) …
(A) Staining for the injected MuSK antibody (10 mg/kg) was evident as early as three days (iv-vi) and persisted for at least seven days after a single injection of antibody (vii-ix). A human …
(A) AChRs are concentrated in the postsynaptic membrane at innervated and fully denervated synapses (scale bar = 20 μm). (B, C) Denervation is evident in female and extensive in male SOD1-G93A mice …
(A) The copy number of the human SOD1-G93A gene was quantified by real-time PCR and normalized to GAPDH. All mice included in this study had 21–26 copies of hSOD1-G93A. The normalized hSOD1-G93A Ct …
(A) The reverse chimera MuSK agonist antibody #13 was injected at P90 () (B) The extent of full innervation decreases progressively from P90 to P162 in female and male SOD1-G93A injected with a …
(A) Human (h) and reverse chimera (rc) MuSK #13 antibodies are similarly effective in stimulating AChR clustering in C2C12 myotubes (n = 3). (B,C) C2C12 myotubes were treated with rc MuSK #13 or a …
Rc MuSK antibody #13 was produced to minimize the occurrence of an immune response to a human antibody, as well as to eliminate the danger of eliciting an immune response at the neuromuscular …
(A) The number (~50%) of partially innervated synapses in SOD1-G93A mice is not altered by chronic injection of reverse chimera MuSK agonist antibody #13 (). The inset shows a representative …
(A,B) 20 Hz stimulation (arrow) of the phrenic nerve from SOD1-G93A mice injected with the control antibody to GP120 led to a rapid and severe decline in the CMAP amplitude. In contrast, the CMAP …
The phrenic nerve was placed in a bipolar suction electrode for stimulation. CMAPs were recorded from a suction recording electrode placed in the same place (dotted circle) across the different …
(A) Representative images of lumbar spinal cords stained with antibodies to ChAT (scale bar = 100 μm). (B) At P138, during the peak period of motor neuron cell death, the number of spinal motor …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
strain, strain background (mouse) | Mouse: B6.Cg-Tg(SOD1*G93A)1Gur/J | The Jackson Laboratory | RRID:IMSR_JAX:004435 | |
strain, strain background (mouse) | Mouse: C57BL/6J | The Jackson Laboratory | RRID:IMSR_JAX000664 | |
cell line (mouse) | Mouse C2C12 skeletal muscle cells | Burden lab | ATCC Cat# CRL-1772, RRID:CVCL_0188 | |
cell line (mouse) | Mouse: Immortalized wild type muscle cells | Burden lab | PMID: 18848351 | |
cell line (mouse) | Mouse: Immortalized Lrp4 mutant muscle cells | Burden lab | PMID: 18848351 | |
antibody | Rabbit anti-Neurofilament-L | SYnaptic SYstems | Cat# 171 002, RRID:AB_887743 | |
antibody | Rabbit anti-Synapsin 1/2 | SYnaptic SYstems | Cat# 106 002, RRID:AB_887804 | |
antibody | Alexa 647-anti-human IgG | Life Technologies | Cat# A-21445, RRID: AB_2535862 | |
antibody | anti-Choline Acetyltransferase | Millipore | Cat# AB144P-200UL, RRID:AB_90661 | |
antibody | anti-NeuN | Millipore | Cat# MAB377, RRID: AB_2298772 | |
antibody | Rabbit anti-MuSK | Burden lab | PMID: 10781064 | |
antibody | Goat anti-MuSK | R and D Systems | Cat# AF3904, RRID:AB_2147242 | |
antibody | Rabbit anti-Dok-7 | Burden lab | PMID: 18848351 | |
antibody | Mouse anti-phosphotyrosine 4G10 | Millipore | ||
peptide, recombinant protein | Alexa 594-alpha-bungarotoxin | Life Technologies | Cat#B13423 | |
peptide, recombinant protein | Alexa 488-alpha-bungarotoxin | Life Technologies | Cat#B13422 | |
peptide, recombinant protein | Hoechst 33342 | Thermo Fisher | Cat# 62249 | |
peptide, recombinant protein | Recombinant Rat Agrin Protein | R and D Systems | Cat#550-AG-100 | |
commercial assay or kit | GAPDH TaqMan Assay Mm00186822_cn | Thermo Fisher | Cat# 4400291 | |
software, algorithm | Prism 7.0 | http://www.graphpad.com/scientific-software/prism/ | RRID:SCR_002798 | |
software, algorithm | Volocity 3D Image Analysis Software | http://www.perkinelmer.com/pages/020/cellularimaging/products/volocity.xhtml | RRID:SCR_002668 | |
software, algorithm | pCLAMP Software Suite | https://www.moleculardevices.com/systems/axon-conventional-patch-clamp/pclamp-11-software-suite | ||
other | DietGel 76A | ClearH20 | Cat#72-07-5022 |