Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Drosophila melanogaster) |
fic | NA | FLYB:FBgn0263278 | |
| Gene (D. melanogaster) |
Hsc3-70 | NA | FLYB:FBgn0001218 | |
| Genetic reagent (D. melanogaster) |
Da-Gal4 | Bloomington Drosophila
Stock Center |
BDSC:55851; FLYB:FBst0055851; RRID: BDSC_55851 |
|
| Genetic reagent (D. melanogaster) |
LongGMR-Gal4 | Bloomington Drosophila
Stock Center |
BDSC:8121; FLYB:FBst0008121; RRID:BDSC_8121 |
|
| Genetic reagent (D. melanogaster) |
W[1118] | Bloomington Drosophila
Stock Center |
BDSC:3605; FLYB:FBst0003605; RRID:BDSC_3605 |
|
| Genetic reagent (D. melanogaster) |
BiP[G0102]/FM7c | Bloomington Drosophila
Stock Center |
BDSC:11815; FLYB:FBal0098203; RRID:BDSC_11815 |
|
| Genetic reagent (D. melanogaster) |
UAS[Scer]-Xbp1- GFP.hg |
Bloomington Drosophila Stock Center |
BDSC:60731; FLYB:FBst0060731; RRID:BDSC_60731 |
Sone et al. (2013) |
| Genetic reagent (D. melanogaster) |
dsRed.crc(ATF4). 5'UTR.tub |
DOI: 10.1371/journal.pone. 0126795; PMID:25978358 |
FLYB: FBal0304834 | Gift from Don Ryoo, NYU. Tubulin promoter and ATF4 5'UTR drive DsRed expression (Flybase FBal0304834) |
| Genetic reagent (D. melanogaster) |
genomic 3xFLAG- BiP[WT] |
This paper | pAttb_gen3xFLAG-BiP[WT] inserted in AttP landing site at 89E11 |
|
| Genetic reagent (D. melanogaster) |
genomic 3xFLAG- BiP[T366A] |
This paper | pAttb_gen3xFLAG-BiP[T366A] inserted in AttP landing site at 89E11 |
|
| Genetic reagent (D. melanogaster) |
genomic 3xFLAG- BiP[T518A] |
This paper | pAttb_gen3xFLAG-BiP[T518A] inserted in AttP landing site at 89E11 |
|
| Genetic reagent (D. melanogaster) |
GMR-dsRNA[white] | This paper | pAttb_GMR-dsRNA[white] inserted in AttP landing site at 43A1 |
|
| Genetic reagent (D. melanogaster) |
fic[30C] | DOI: 10.1074/jbc.M117. 799296; PMID:29089387 |
Casey et al. (2017) | |
| Genetic reagent (D. melanogaster) |
UAS[Scer]-V5- Fic[E247G] |
DOI: 10.1074/jbc.M117. 799296; PMID:29089387 |
Casey et al. (2017) | |
| Genetic reagent (D. melanogaster) |
UPR and ER protein RNAi lines screened are contained in Supplemental Table 1 |
|||
| Genetic reagent (Saccharomyces cerevisiae) |
KAR2::KAN | GE Healthcare Life Sciences |
SGD:S000003571 | gene replacement generated using PCR-based gene deletion strategy yielding start- to stop-codon deletion |
| Strain, strain background (Saccharomyces cerevisiae) |
BY4741 MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 |
DOI: 10.1002/(SICI)1097- 0061(19980130)14:2 < 115:: AID-YEA204 > 3.0.CO;2–2; PMID: 9483801 |
GenBank: JRIS00000000.1 | |
| Antibody | anti-Hsc70-3 (BiP) (Guinea Pig polyclonal) |
DOI: 10.1038/sj.emboj. 7601477; PMID:17170705 |
FLYB: FBgn0001218; RRID: AB_2569409 |
Gift from Don Ryoo, NYU (1:2000 IHC, 1:8000 WB) |
| Antibody | anti-RFP (Rabbit polyclonal) |
Rockland | Rockland:600-401-379; RRID:AB_2209751 |
(1:1000 IHC) |
| Antibody | anti-GFP (Chicken polyclonal) |
ThermoFisher Scientific |
ThermoFisher Scientific:A10262; RRID: AB_2534023 |
(1:1000 IHC) |
| Antibody | anti-Flag (mouse monoclonal) |
Sigma | Sigma:F-3165; RRID:AB_259529 |
(1:2000 WB) |
| Antibody | anti-Actin (mouse monoclonal) |
Developmental Studies Hybridoma Bank |
DSHB:JLA20; RRID: AB_528068 |
1:2000 (WB) |
| Antibody | Alexa 488- or 568- secondaries |
Molecular Probes | (1:1000 IHC) | |
| Antibody | LICOR 800 or 700- secondaries |
LICOR Biosciences | (1:20,000 WB) | |
| Recombinant DNA reagent |
pAttb_gen3xFLAG- BiP[WT] |
This paper | PCR in multiple steps from genomic DNA {sequence location = X: 11,801,696..11,807,117 [-]}. Cloned into modified pAttb vector |
|
| Recombinant DNA reagent |
pAttb_gen3xFLAG- BiP[T366A] |
This paper | Progenitors: pAttb_gen3xFLAG- BiP[WT]. Mutated sequence synthesized with Geneblock (IDT) |
|
| Recombinant DNA reagent |
pAttb_gen3xFLAG- BiP[T518A] |
This paper | Progenitors: pAttb_gen3xFLAG- BiP[WT]. Mutated sequence synthesized with Geneblock (IDT) |
|
| Recombinant DNA reagent |
pAttb_GMR- dsRNA[white] |
This paper | Progenitors: pUASt_dsRNA[white] (gift from Dean Smith, UT Southwestern, PMID: 11804566). GMR sequence: Geneblock (IDT) |
|
| Recombinant DNA reagent |
pKAR2:LEU2 | This paper | Cloned from amplification of endogenous KAR2 with primers (see below) |
|
| Recombinant DNA reagent |
pKAR2[T386A]: LEU2 |
This paper | Progenitor: pKAR2:LEU2. Site directed mutagenesis used to make mutation |
|
| Recombinant DNA reagent |
pKAR2[T538A]: LEU2 |
This paper | Progenitor: pKAR2:LEU2. Site directed mutagenesis used to make mutation |
|
| Sequence-based reagent |
5’-GCATCCGCGGATACT CTCGTACCCTGCCGC-3’ |
This paper | Cloning for pKAR2:LEU2 | |
| Sequence-based reagent |
5’-ATGCGAGCTCCGTAT ATACTCAGTATAATC-3’ |
This paper | Cloning for pKAR2:LEU2 | |
| Sequence-based reagent |
5’-GGTTGGTGGTTCTG CTAGAATTCCAAAGGT CCAACAATTGTTAGAA TCATACTTTGATGG-3’ |
This paper | Mutagenesis primer for pKAR2[T386A]:LEU2 |
|
| Sequence-based reagent |
5’-ACCTTTGGAATTCT AGCAGAACCACCAAC CAAAACGATATCATCA ACATCCTTCTTTTCC-3’. |
This paper | Mutagenesis primer for pKAR2[T386A]:LEU2 |
|
| Sequence-based reagent |
5’-AGATAAGGGAGCTGG TAAATCCGAATCTATCAC CATCACTAACG-3’ |
This paper | Mutagenesis primer for pKAR2[T538A]:LEU2 |
|
| Sequence-based reagent |
5’-GGATTTACCAGCTCC CTTATCTGTGGCAGACA CCTTCAGAATACC-3’. |
This paper | Mutagenesis primer for pKAR2[T538A]:LEU2 |
|
| Chemical compound, drug |
VECTASHIELD Antifade Mounting Medium with DAPI |
Vector Laboratories | Vector Laboratories: H-1200 |
|
| Software, algorithm | Adobe Photoshop | Adobe | RRID:SCR_014199 | |
| Software, algorithm | ImageJ | NIH | RRID:SCR_003070 |