KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit anti-IRE1α | Cell Signaling | Cat. #3294; RRID:AB_823545 |
| Rabbit anti-IRE1α | Santa Cruz | Cat. #sc-20790; RRID:AB_2098712 |
| Mouse anti-Tubulin | Abcam | Cat. #ab7291; RRID:AB_2241126 |
| Rabbit anti-BiP | Cell Signaling | Cat. #3177; RRID:AB_2119845 |
| Mouse anti-BiP | BD Biosciences | Cat. #610979; RRID:AB_398292 |
| Rabbit anti-ATF6α | Cell Signaling | Cat. #65880; RRID:AB_2799696 |
| Rabbit anti-PERK | Cell Signaling | Cat. #3192; RRID:AB_2095847 |
| Goat anti-mouse IgG-HRP | Jackson Immunoreserach | Cat. #115-035-003; RRID:AB_10015289 |
| Goat anti-rabbit IgG-HRP | Jackson Immunoreserach | Cat. #111-035-003; RRID:AB_2313567 |
| Goat anti-rabbit IgG-Cy3 | Jackson Immuno Research | Cat. #711-165-152; RRID:AB_2307443 |
| Rabbit anti-Sec61α | Gift from Dr. Ramanujan Hegde | N/A |
| Rabbit anti-Sec62 | Gift from Dr. Ramanujan Hegde | N/A |
| Rabbit anti-Sec63 | Gift from Dr. Ramanujan Hegde | N/A |
| Rabbit anti-HA | Gift from Dr. Ramanujan Hegde | N/A |
| Rabbit anti-Sec61α | Gift from Dr. Ramanujan Hegde | N/A |
| Resins | ||
| mouse anti-HA magnetic beads | Fisher Scientific | 88836 |
| Rat anti-FLAG beads | Biolegend | 651503 |
| StrepTactin-XT Sepharose | IBA, Germany | 2-1201-010 |
| Protein A agarose | Repligen | CA-PRI-1000 |
| Reagents | ||
| DMEM | Corning | 10–013-CV |
| FBS | GIBCO | 16000044 |
| Horse Serum | Sigma | H0146 |
| Penicillin/Streptomycin | GIBCO | 15140122 |
| Lipofectamine 2000 | Invitrogen | 11668019 |
| Doxycycline | Clontech | 631311 |
| Hygromycin B | Invitrogen | 10687010 |
| Blasticidin | InvivoGen | ant-bl-1 |
| Thapsigargin | Enzo Life Sciences | BML-PE180 |
| Protease inhibitor cocktail | Roche | 11873580001 |
| Poly L-lysine | Peptides International | OKK-3056 |
| Digitonin | EMD Millipore | 300410 |
| Fluoromount G | SouthernBiotech | 0100–01 |
| Phos-tag | Wako | 300–93523 |
| West Pico Substrate | Thermo Scientific | 34080 |
| Femto Substrate | Thermo Scientific | 34095 |
| Oligo(dT) primers | QIAGEN | 79237 |
| M-MLV reverse transcripts | NEB | M0253 |
| Trizol reagent | Thermo Scientific | 15596026 |
| DMSO | Sigma-Aldrich | D2650 |
| Tunicamycin | Sigma- Aldrich | T7765 |
| DTT | AmaricanBio | 3483-12-2 |
| PonceauS solution | Sigma-Aldrich | P3504 |
| Sec62 siRNA | QIAGEN | 68875 |
| Sec63 siRNA | QIAGEN | 68711 and 68715 |
| Experimental Models: Cell line | ||
| HEK293-Flp-In T-Rex cells | Invitrogen | R78007 |
| HEK293-Flp-In Sec63−/− | Sun and Mariappan, 2020 | N/A |
| HEK293-Flp-In Sec62−/− | This study | N/A |
| HEK293-Flp-In Sec63−/−+Sec63-FLAG | Sun and Mariappan, 2020 | N/A |
| HEK293-Flp-In Sec63−/−+Sec63 J-mutant-FLAG | This study | N/A |
| HEK293-Flp-In IRE1α−/−+IRE1α-HA | Plumb et al., 2015 | N/A |
| HEK293-Flp-In IRE1α−/−+IRE1α-CNX-TMD-HA | This study | N/A |
| 2xStrep-Sec61α | This study | N/A |
| Recombinant DNA | ||
| pOG44 | Invitrogen | V600520 |
| pSpCas9(BB)-2A-Puro | Addgene | 62988 |
| pCR-BluntII | Addgene | K2800-20 |
| pCDNA5/FRT/TO-empty vector | Invitrogen | V652020 |
| pCDNA5/FRT/TO-Sec63-Flag | Sun and Mariappan, 2020 | N/A |
| pCDNA5/FRT/TO-Sec63 J mut-Flag | This study | N/A |
| pCDNA5/FRT/TO-IRE1α -HA | Plumb et al., 2015 | N/A |
| pCDNA5/FRT/TO-IRE1α -CNX-TMD-HA | This study | N/A |
| pCDNA5/FRT/TO-IRE1α-VD/AA -HA | Plumb et al., 2015 | N/A |
| pCDNA5/FRT/TO-IRE1α-ΔLD -HA | Plumb et al., 2015 | N/A |
| pCDNA5/FRT/TO-IRE1α-M/A -HA | Plumb et al., 2015 | N/A |
| pCDNA5/FRT/TO-IRE1α-L/A -HA | Plumb et al., 2015 | N/A |
| pCDNA5/FRT/TO- Sec63 Δ367-760-FLAG | This study | N/A |
| pCDNA5/FRT/TO- Sec63 Δ637-760-FLAG | This study | N/A |
| pCDNA5/FRT/TO- Sec63 Δ230-300-FLAG | This study | N/A |
| pCDNA5/FRT/TO- Sec63 Δ230-760-FLAG | This study | N/A |
| Software and Algorithms | ||
| imageJ | https://imagej.nih.gov/ij/ | N/A |
| Graphpad | https://graphpad.com:443/scientific-software/prism/ | N/A |