Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference |
Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | FBXW5 | This paper | Yenzyme | (1:250 dilution for western blotting) |
| Antibody | Phospho- SEC23B (S186) |
This paper | Yenzyme | (1:200 dilution for western blotting) |
| Antibody | SEC23A | Randy Schekman’s lab |
PMID: 24069399 | (1:1000 dilution for western blotting) |
| Antibody | SEC23B | Randy Schekman’s lab |
PMID: 24069399 | (1:1000 dilution for western blotting) |
| Antibody | SEC24A | Randy Schekman’s lab |
PMID: 18056412 | (1:2000 dilution for western blotting) |
| Antibody | SEC24B | Randy Schekman’s lab |
PMID: 18056412 | (1:2000 dilution for western blotting) |
| Antibody | SEC24C | Randy Schekman’s lab |
PMID: 18056412 | (1:2000 dilution for western blotting) |
| Antibody | SEC24D | Randy Schekman’s lab |
PMID: 18056412 | (1:2000 dilution for western blotting) |
| Antibody | FLAG | Sigma-Aldrich | Cat# F7425 | (1:1000 dilution for western blotting, 1:1000 for immunofluorescence) |
| Antibody | HA | Covance | Cat# MMS- 101P |
(1:1000 dilution for western blotting) |
| Antibody | MYC | Sigma-Aldrich |
Cat# M5546 |
(1:1000 dilution for western blotting) |
| Antibody | SAR1 | Thermo Scientific |
Cat# PA5 -27642 |
(1:5000 dilution for western blotting) |
| Antibody | CUL1 | Life Technologies |
Cat# 322400 | (1:1000 dilution for western blotting) |
| Antibody | SKP1 | Michele Pagano’s lab |
Yenzyme | (1:1000 dilution for western blotting) |
| Antibody | SEC13 | Bethyl Laboratories |
Cat# A303 -980A |
(1:10,000 dilution for western blotting) |
| Antibody | SEC31A | Bethyl Laboratories |
Cat# A302 -336A |
(1:10,000 dilution for western blotting) |
| Antibody | ULK1 | Cell Signaling Technology |
Cat# 8054 s | (1:1000 dilution for western blotting) |
| Antibody | Phospho-ULK1 (S757) | Cell Signaling Technology |
Cat# 6888 s | (1:1000 dilution for western blotting) |
| Antibody | LC3B | Novus Biologicals |
Cat# NB100 -2220 |
(1:10,000 dilution for western blotting, 1:1000 for immunofluorescence) |
| Antibody | Phospho- ATG13 (S318) |
Rockland Immunochemicals |
Cat# 600– 401 C49S |
(1:1000 dilution for western blotting) |
| Antibody | Phospho- Beclin-1 (S15) |
Abbiotec | Cat# 254515 | (1:500 dilution for western blotting) |
| Antibody | Beclin-1 | Santa Cruz Biotechnology |
Cat# SC-48381 | (1:1000 dilution for western blotting) |
| Antibody | GAPDH | Cell Signaling Technology |
Cat# 97166S | (1:10,000 dilution for western blotting) |
| Antibody | ACTIN | Sigma-Aldrich | Cat# A5441 | (1:10,000 dilution for western blotting) |
| Antibody | Tubulin | Sigma-Aldrich | Cat# T6074 -200UL |
(1:10,000 dilution for western blotting) |
| Antibody | ERGIC53 | Sigma-Aldrich | Cat# E1031 -200UL |
(1:5000 dilution for western blotting, 1:1000 for immunofluorescence) |
| Antibody | SEC16 | Bethyl Laboratories |
Cat# A300-648A | (1:5000 dilution for western blotting) |
| Antibody | FBXO11 | Novus Biologicals |
Cat# H00080204 -B01 |
(1:1000 dilution for western blotting) |
| Antibody | FBXW9 | Michele Pagano’s lab |
(1:1000 dilution for western blotting) |
|
| Peptide, recombinant protein |
MG132 | Peptide international |
Cat# IZL-3175v | (final concentration, 5 mM) |
| Chemical compound, drug |
MLN4924 | Active Biochem | Cat# A-1139 | (final concentration, 1 mM) |
| Chemical compound, drug |
Cycloheximide | Sigma-Aldrich | Cat# C7698-1G | (final concentration, 50 ng/ml) |
| Chemical compound, drug |
Bafilomycin A1 | Santa Cruz Biotechnology |
Cat# sc-201550A | (final concentration, 0.1 mg/ml) |
| Chemical compound, drug |
Polybrene | Sigma-Aldrich | Cat# H9268-10G | (final concentration, 8 μg/ml) |
| Chemical compound, drug |
SBI-0206965 | Selleck Chemicals |
Cat# S7885 | (final concentration, 1 mM) |
| Peptide, recombinant protein |
SEC23B peptide |
This paper | Yenzyme | |
| Peptide, recombinant protein |
SEC23B phospho- peptide |
This paper | Yenzyme | |
| Cell line (human) | HEK293T | ATCC | Cat# CRL-3216 | |
| Cell line (human) | U-2OS | ATCC | Cat# HTB-96 | |
| Cell line (human) | RPE1-hTERT | ATCC | Cat# CRL-4000 | |
| Cell line (human) | A375 | ATCC | Cat# CRL-1619 | |
| Cell line (human) | HeLa | ATCC | Cat# CCL-2 | |
| Commercial assay or kit |
Pierce Gaussia Luciferase Flash Assay Kit |
Thermo Scientific |
Cat# 16159 | |
| Other | siRNAs to FBXW5 (#1) |
Dharmacon | Cat# J-013389 -05-0002 |
Oligonucleotides |
| Other | siRNAs to FBXW5 (#2) |
Dharmacon | Cat# J-013389 -06-0002 |
Oligonucleotides |
| Other | siRNAs to FBXW5 (#3) |
Dharmacon | Cat# J-013389 -07-0002 |
Oligonucleotides |
| Other | siRNAs to FBXW5 (#4) |
Dharmacon | Cat# J-013389 -08-0002 |
Oligonucleotides |
| Other | siRNAs to FBXW5 |
Santa Cruz Biotechnology |
Cat# sc-92629 | Oligonucleotides |
| Other | siRNAs to SEC23A |
Dharmacon | Cat# M-009582 -00-0005 |
Oligonucleotides |
| Other | siRNAs to SEC23B |
Dharmacon | Cat# M-009592 -01-0005 |
Oligonucleotides |
| Other | siRNAs to ULK1 |
Santa Cruz Biotechnology |
Cat# sc-44182 | Oligonucleotides |
| Other | siRNAs to ULK2 |
Santa Cruz Biotechnology |
Cat# sc-44183 | Oligonucleotides |
| Other | siRNAs to SEC24A |
Dharmacon | Cat# L-024405 -01-0005 |
Oligonucleotides |
| Other | siRNAs to SEC24B |
Dharmacon | Cat# L-008299 -02-0005 |
Oligonucleotides |
| Other | siRNAs to SEC24C |
Dharmacon | Cat# L-008467 -02-0005 |
Oligonucleotides |
| Other | siRNAs to SEC24D |
Dharmacon | Cat# L-008493 -01-0005 |
Oligonucleotides |
| Other | Non-targeting siRNA (CGUACGCGGAAUACUUCGA) |
Dharmacon | Oligonucleotides | |
| Recombinant DNA reagent |
pCS2 + 3x HA-Sec24A |
Randy Schekman’s lab |
||
| Recombinant DNA reagent |
pCS2 + 3x HA-Sec24B |
Randy Schekman’s lab |
||
| Recombinant DNA reagent |
pCS2 + 3x HA-Sec24C |
Randy Schekman’s lab |
||
| Recombinant DNA reagent |
pCS2 + 3x HA-Sec24D |
Randy Schekman’s lab |
||
| Recombinant DNA reagent |
pBabe-puro- mCherry-EGFP-LC3B |
Addgene | Cat# 22418 | |
| Recombinant DNA reagent |
pcdna6.2- myc ULK1 wt |
Addgene | Cat# 27629 | |
| Recombinant DNA reagent |
pcdna6.2- myc ULK1 k46I |
Addgene | Cat# 27630 | |
| Recombinant DNA reagent |
GFP-SAR1 (T39N) |
Antonella De Matteis’ lab |
||
| Recombinant DNA reagent |
GFP-SAR1 (H79G) |
Antonella De Matteis’ lab |