Table 1. List of reagents.
| Reagent or resource | Source | Identifier |
|---|---|---|
| Antibodies | ||
| Rat anti-galectin-3-647 mAbFlow cytometry
concentration 50 µg ml -1 |
BioLegend | 125408, RRID: AB_1186110 |
| Sheep antiTGN46 pAb, IF concentration 5 µg ml -1 | BioRad | AHP500, RRID: AB_324049 |
| Mouse anti-GM130 mAb, IF concentration 25 µg ml -1 | BD Bioscienes | 610822, RRID: AB_398141 |
| Mouse anti-Sec31 mAb, IF concentration 25 µg ml -1 | BD Bioscienes | 612351, RRID: AB_39971 |
| Mouse anti-MHC-I mAb, IF concentration 20 µg ml -1 | ThermoFisher | MA1-70111, RRID: AB_1076705 |
| Rabbit anti-calnexin pAb, IF concentration 5 µg ml -1 | Abcam | ab22595, RRID: AB_2069006 |
| Rabbit anti-GPR161 pAb, IF concentration 10 µg ml -1 | MyBioSource | MBS719938, RRID: AB_2801289 |
| Rabbit anti-TMEM220 pAb, IF concentration 100 µg ml -1 | Aviva Systems Biology | ARP44467_ P050, RRID:
AB_2801328 |
| Rabbit anti-Golgin A5 pAb, IF concentration 10 µg ml -1 | GeneTex | GTX104255, RRID: AB_2037117 |
| Donkey anti-sheep IgG-568 pAb, IF concentration
4 µg ml -1 |
ThermoFisher | A21099, RRID: AB_2535753 |
| Goat anti-mouse IgG-488 pAb, IF concentration
4 µg ml -1 |
ThermoFisher | A11001, RRID: AB_2534069 |
| Goat anti-rabbit IgG-568 pAb, IF concentration
4 µg ml -1 |
ThermoFisher | A11011, RRID: AB_143157 |
| Mouse anti-FLAG M2 mAb, IF concentration 0.5 µg ml -1 | Sigma | F1804, RRID: AB_262044 |
| Cell lines | ||
| HeLa | ATCC | CCL-2 |
| HeLa-HRP | Gift from Vivek Malhotra | As described 20 |
| sHeLa (HeLa S3) | Gift from Paul Lehner | Sigma, 87110901 |
| sHeLa-Cas9 | In house production | As described 21 |
| Genome-wide screen library | ||
| Lentiviral Brunello | David Root and John Doench, Addgene | 73178-LV |
| Plasmids | ||
| Mouse-GPR161-FLAG | Insight Biotechnology LTD | MR224248 |
| Human-TMEM220-FLAG | Genescript USA INC | OHu14539 |
| Human-COL14A1-FLAG | Stratech Scientific LTD | HG13622-CF-SIB |
| Mouse-FAM98B-FLAG | Insight Biotechnology LTD | MR206836 |
| Mouse-FAM102B-FLAG | Insight Biotechnology LTD | MR212923 |
| Mouse-MXRA7-FLAG | Insight Biotechnology LTD | MR218215 |
| Software | ||
| MaGECK | https://sourceforge.net/p/mageck/wiki/Home/ | Version 0.5.7 |
| MaGECK-vispr | https://bitbucket.org/liulab/mageck-vispr | Version 0.5.3 |
| GraphPad PRISM | GraphPad | Version 7 |
| GO term mapper | https://go.princeton.edu/cgi-bin/GOTermMapper | Accessed 7 th Jan 2019 |
| CellProfiler | http://cellprofiler.org/releases/ | Version 3.1.5 |
| CellProfiler Analyst | http://cellprofiler.org/releases/ | Version 2.2.1 |
| Fiji | https://imagej.net/Fiji/Downloads | ImageJ-win64, downloaded 25th
May 2018 |
| R | https://www.r-project.org/ | Version 3.5.1 |
| Reagents | ||
| High glucose Dulbecco’s Modified Eagle Medium
(DMEM) |
Molecular Probes | D6546 |
| Hank's Balanced Salt Solution (HBSS) | ThermoFisher | 14025092 |
| Fetal bovine serum (FBS) | ThermoFisher | 10270-106 |
| L-glutamine | Sigma | G7513 |
| Penicillin/streptomycin | Sigma | P0781 |
| Polybrene | Merck | TR-1003-G |
| Dulbecco’s Modified Eagle Medium (DMEM) | Life Technologies | 21063-029 |
| Propidium iodide | Biolegend | 421301 |
| Gentra Puregene Cell kit | Qiagen | 158388 |
| dNTP | ThermoFisher | R0191 |
| ExTaq | Takara | RR001B |
| AMPure XP magnetic beads | Beckman Coulter | A63880 |
| White 96 well CulturPlate | PerkinElmer | 6005680 |
| 96 well ViewPlate | PerkinElmer | 6005182 |
| PBS | ThermoFisher | D8537 |
| ProLong with DAPI | Invitrogen | P36962 |
| Paraformaldehyde | Sigma | 158127 |
| TransIT 2020 | Mirus | MIR 5405 |
| Lipofectamine 2000 | ThermoFisher | 12566014 |
| Phalloidin Green | ThermoFisher | A12379 |
| ProLong Gold with DAPI | ThermoFisher | P36935 |
| Tween 20 | Sigma | P1379 |
| ECL Western Blotting Detection Reagent | Amersham | RPN2106 |
| Westar XLS100 | Cyanagen | XLS100 |
| Restore Plus | ThermoFisher | 46430 |
| Primers and siRNA | ||
| P5 primer mix | Integrated DNA Technologies | See Extended data, E2 22 |
| P7 primers | Integrated DNA Technologies | See Extended data, E3 22 |
| siGenome smart pool plate | Dharmacon | G-CUSTOM-384690
(see Extended data, E4) 22 |