Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) |
Rapsn-/- | PMID: 7675108 | ||
| Genetic reagent (M. musculus) |
N88K | This paper | See detail information in Materials and methods . | |
| Cell line (Homo sapiens) | HEK293T | ATCC | Cat#:CRL-3216 RRID: CVCL_0042 |
From ATCC; Cell identity has been confirmed by STR profiling and cell line was found to be free of Mycoplasma. |
| Cell line (M. musculus) | C2C12 | ATCC | Cat#: CRL-1772 RRID: CVCL_0188 |
From ATCC; Cell identity has been confirmed by STR profiling and cell line was found to be free of Mycoplasma. |
| Cell line (M. musculus) | N88K mt C2C12 | This paper | See detail information in Material and methods). | |
| Cell line (M. musculus) | Y86F mt C2C12 | This paper | See detail information in Material and methods. | |
| Cell line (M. musculus) | Rapsn-/- C2C12, clone 11–7 | PMID: 10414969 | ||
| Antibody | Mouse monoclonal anti-HA Agarose | Thermo Fisher Scientific | Cat#: 26181, RRID: AB_2537081 |
1: 40 for IP |
| Antibody | Goat anti-rabbit IgG conjugated with Alexa Fluor 488 | Thermo Fisher Scientific | Cat#: A-11008 RRID: AB_10563748 |
IHC (1:500) |
| Antibody | Rabbit polyclonal anti-Actin | Cell Signaling Technology | Cat #: 4967, RRID: AB_330288 |
WB (1:10000) |
| Antibody | Rabbit polyclonal anti- α-Actin | Abcam | Cat #: ab52218 RRID: AB_870573 |
WB (1:1000) |
| Antibody | Rabbit polyclonal anti-Flag | Sigma-Aldrich | Cat #: F7425 RRID: AB_439687 |
WB (1:1000) |
| Antibody | Mouse monoclonal anti-GAPDH (Clone, 6C5) | Santa Cruz Biotechnology | Cat #: sc-32233, RRID: AB_627679 |
WB (1:10000) |
| Antibody | Rabbit polyclonal anti-neurofilament | Cell Signaling Technology | Cat#: 2837, RRID: AB_823575 |
IHC (1:1000) |
| Antibody | Rabbit polyclonal anti-synapsin | Cell Signaling Technology | Cat#: 5297, RRID: AB_2616578 |
IHC (1:1000) |
| Antibody | Mouse monoclonal anti-Ubiquitin (Clone, Ubi-1) | Abcam | Cat#: ab7254, RRID: AB_305802 |
WB (1:1000) |
| Antibody | Rabbit polyclonal anti-GFP | Cell Signaling Technology | Cat#: 2555, RRID: AB_10692764 |
WB (1:1000) |
| Antibody | Mouse monoclonal P-Tyr-100 | Cell Signaling Technology | Cat #: 9411 RRID: AB_331228 |
WB (1:1000) |
| Antibody | Mouse monoclonal Anti-Flag affinity gel (Clone, M2) | Sigma-Aldrich | Cat#: A2220, RRID: AB_10063035 |
1: 50 for IP |
| Antibody | Rabbit polyclonal anti-HA | Sigma-Aldrich | Cat#: H6908, RRID: AB_260070 |
WB (1:2000) |
| Antibody | Mouse monoclonal anti-Rapsn (Clone 1234) | Abcam | Cat#: ab11423, RRID: AB_298028 |
WB (1:1000) |
| Antibody | Rabbit polyclonal anti-Transferrin | Abcam | Cat#: ab82411, RRID: AB_1659060 |
WB (1:1000) |
| Antibody | Mouse monoclonal anti-δ-AChR (Clone, 88B) | Thermo Fisher Scientific | Cat#: MA3-043, RRID: AB_2081037 |
WB (1:1000) |
| Antibody | Rabbit polyclonal anti-Rapsn | PMID: 18940591 | WB (1:1000) | |
| Antibody | Goat polyclonal anti-α-AChR | PMID: 3484485 | WB (1:1000) | |
| Antibody | Rabbit polyclonal anti-β-AChR | PMID: 3484485 | WB (1:1000) | |
| Antibody | Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG | Thermo Fisher Scientific | Cat#: 32260, RRID: AB_1965959 |
WB (1:5000) |
| Antibody | Horseradish peroxidase (HRP)-conjugated goat anti-Mouse IgG | Thermo Fisher Scientific | Cat#: 32230, RRID: AB_1965958 |
WB (1:5000) |
| Antibody | Horseradish peroxidase (HRP)-conjugated goat anti-Rat IgG | Thermo Fisher Scientific | Cat#: 31470, RRID: AB_228356 |
WB (1:5000) |
| Recombinant Protein | Agrin | R and D Systems | Cat#: 550-AG-100 | 50 ng / ml for induction of AChR clusters in culture myotubes |
| Other | Immunofluorescence of NMJ in diaphragm and muscles | PMID: 18278041
PMID: 22794264 |
||
| Other | Electron microscopic analysis | |||
| Other | Generation of Gene-modified C2C12 cells | PMID: 27839998 | ||
| Other | Isolation of cell surface protein | PMID: 22157653 | ||
| Other | Isolation of surface AChR and their associated proteins | PMID: 18940591 | ||
| Other | AAV Virus production | PMID: 30626963 and Information in Addgene: https://www.addgene.org/protocols/aav-production-hek293-cells/ |