Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | emerin | NCBI RefSeq NM_007927 | ||
| Gene (Mus musculus) | nurim | NCBI RefSeq NM_134122 | ||
| Gene (Mus musculus) | Sun2 | NCBI RefSeq NM_001205346 | ||
| Cell line (Mus musculus) | C2C12 | ATCC | CRL-1772 | |
| Cell line (Homo sapiens) | U-2-OS | ATCC | HTB-96 | |
| Recombinant DNA reagent (plasmid) | pQCXIB vector | Campeau et al. (2009) Addgene | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | Myc/FLAG RITE vector | Toyama et al. (2019) | Lentiviral contruct for stable expression of RITE-tagged protein | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-GFP | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-D95-99-GFP | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-DLEM-GFP | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-GFP-SSNKTVD | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-Δ95–99-GFP-SSNKTVD | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB emerin-ΔLEM-GFP-SSNKTVD | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB Sun2-GFP | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | pQCXIB nurim-GFP | This paper | Retroviral construct for stable expression | |
| Recombinant DNA reagent (plasmid) | Emerin-RITE | This paper | Lentiviral contruct for stable expression of RITE-tagged protein | |
| Recombinant DNA reagent (plasmid) | Nurim-RITE | This paper | Lentiviral contruct for stable expression of RITE-tagged protein | |
| Recombinant DNA reagent (plasmid) | Emerin-Δ95–99-RITE | This paper | Lentiviral contruct for stable expression of RITE-tagged protein | |
| Antibody | Rabbit polyclonal anti-emerin | Santa Cruz Biotechnology | Sc-15378 | WB (1:1000) |
| Antibody | GFP | Abcam | ab290 | Ab uptake (1:500); WB (1:1000) |
| Antibody | Mouse monoclonal anti-FLAG | Sigma-Aldrich | F1804 | IF (1:1000) |
| Antibody | Mouse monoclonal anti-Myc | Cell Signaling | 2233 | IF (1:1000); Ab uptake (1:500) |
| Antibody | Mouse monoclonal anti-tubulin | Sigma-Aldrich | T5168 | WB (1:2500) |
| Antibody | giantin | BioLegend | PRB-114C | IF (1:1000) |
| Antibody | LAMP1 | Abcam | ab24170 | IF(1:100) |
| Other | Alexa-647 WGA | Life Technologies | W32466 | IF (5 ug/ml) |
| Commercial assay or kit | PNGase F | NEB | P0704 | |
| Commercial assay or kit | Endo H | NEB | P0702 | |
| Chemical compound, drug | Thapsigargin | Thermo Fisher | T7459 | Used at 100 nM |
| Chemical compound, drug | MG132 | Cayman Chemical | 1211877-36-9 | Used at10 uM |
| Chemical compound, drug | Bafilomycin A1 | BioViotica | BVT-0252 | Used at100 nM |
| Chemical compound, drug | Brefeldin A | Tocris | 1231 | Used at2.5 uM |
| Chemical compound, drug | Leupeptin | Sigma-Aldrich | L5793 | Used at 125 uM |
| Chemical compound, drug | cycloheximide | Sigma-Aldrich | C-7698 | Used at200 ug/ml |
| Other | 13C6-Lysine | Cambridge Isotopes | CLM-2247 | |
| Other | 13C6, 15N4-Arginine | Cambridge Isotopes | CNLM-539 | |
| Other | Lysine/arginine free DMEM | Thermo Fisher | 88364 | |
| Other | Dialyzed fetal bovine serum | Thermo Fisher | 26400044 | |
| Other | Hoechst stain | Molecular Probes | H1399 | Used at 10 ug/ml |
| Recombinant DNA reagent (plasmid) | UBE2G1 miR-E LT3GEPIR | Knott et al., 2014 | TGCTGTTGACAGTGAGCGAAAGACAGCTGGCAGAACTCAATAGTGAAGCCACAGATGTATTGAGTTCTGCCAGCTGTCTTCTGCCTACTGCCTCGGA | |
| Recombinant DNA reagent (plasmid) | UBE2G2 miR-E LT3GEPIR | Knott et al., 2014 | TGCTGTTGACAGTGAGCGAACCGGGAGCAGTTCTATAAGATAGTGAAGCCACAGATGTATCTTATAGAACTGCTCCCGGTCTGCCTACTGCCTCGGA | |
| Recombinant DNA reagent (plasmid) | UBE2J1 miR-E LT3GEPIR | Knott et al., 2014 | TGCTGTTGACAGTGAGCGAAAGGTTGTCTACTTCACCAGATAGTGAAGCCACAGATGTATCTGGTGAAGTAGACAACCTTCTGCCTACTGCCTCGGA | |
| Recombinant DNA reagent (plasmid) | MARCH6 miR-E LT3GEPIR | Knott et al., 2014 | TGCTGTTGACAGTGAGCGACTGGATCTTCATTCTTATTTATAGTGAAGCCACAGATGTATAAATAAGAATGAAGATCCAGCTGCCTACTGCCTCGGA | |
| Software, algorithm | Fiji | https://fiji.sc/ | ||
| Software, algorithm | RStudio | https://rstudio.com/ |