Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo-sapiens) | HEK293, Human embryonic kidney cells | ATCC | CRL-3216 | corresponding Hsp90α/β KO cell lines were generated |
| Cell line (Homo-sapiens) | A549, human lung epithelial carcinoma cells | ATCC | CCL-185 | corresponding Hsp90α/β KO cell lines were generated |
| Cell line (Homo-sapiens) | RPE1, human retinal epithelial cells | ATCC | CRL-4000 | |
| Cell line (Homo-sapiens) | HCT116, human colon carcinoma Cells | ATCC | CCL-247 | |
| Cell line (Mus musculus) | Mouse adult fibroblasts (MAFs) | Bhattacharya et al., 2022 | ||
| Recombinant DNA reagent | pcDNA-Flag HSF1 wt (plasmid) | a gift from Len Neckers (Kijima et al., 2018) | transient overexpression of WT Hsf1 | |
| Recombinant DNA reagent | pcDNA-Flag HSF1 C205 (plasmid) | a gift from Len Neckers (Kijima et al., 2018) | This express the Hsf1 mutant comprising only the N-terminal 205 amino acids | |
| Recombinant DNA reagent | pcDNA3.1(+) (plasmid) | Thermo Fisher Scientific | #V79020 | |
| Recombinant DNA reagent | pCherry.90α (plasmid) | Picard et al., 2006 | transient overexpression of Hsp90α | |
| Recombinant DNA reagent | pEGFP.90β (plasmid) | Picard et al., 2006 | transient overexpression of Hsp90β | |
| Recombinant DNA reagent | pmCherry-C1 (plasmid) | Picard et al., 2006 | Control for pCherry.90α | |
| Recombinant DNA reagent | pEGFP-C1 (plasmid) | Clontech | #6084–1 | |
| Recombinant DNA reagent | FUW mCherry-GFP-LC3 (plasmid) | a gift from Anne Brunet (Leeman et al., 2018) | Addgene #110060 | autophagy reporter plasmid |
| Recombinant DNA reagent | Ub-M-GFP (plasmid) and Ub-R-GFP (plasmid) |
a gift from Nico Dantuma (Dantuma et al., 2000) | Addgene #11938 and #11939 | |
| Recombinant DNA reagent | HSE (WT)-Luc (plasmid) | a gift from Ueli Schibler (Reinke et al., 2008) | Hsf1 reporter plasmid | |
| Recombinant DNA reagent | pEGFP-Q74 (plasmid) | a gift from David Rubinsztein (Narain et al., 1999) | Addgene # 40261 | |
| Recombinant DNA reagent | pRL-CMV (plasmid) | Promega | #E2261 | |
| Recombinant DNA reagent | pSpCas9(BB)–2A-Puro (PX459) (plasmid) | a gift from Feng Zhang (Ran et al., 2013) | Addgene #48139 | |
| Recombinant DNA reagent | pGL3-CMV.Luc (plasmid) | a gift from Laurent Guillemot (University of Geneva) | ||
| Sequence-based reagent | shHSF1-1 | This paper, from Microsynth | HSF1 shRNA 3UTR forward | 5'-CCG GGC AGG TTG TTC ATA GTC AGA ACT CGA GTT CTG ACT ATG AAC AAC CTG CTT TTT G-3' |
| Sequence-based reagent | shHSF1-1 | This paper, from Microsynth | HSF1 shRNA 3UTR reverse | 5'-AAT TCA AAA AGC AGG TTG TTC ATA GTC AGA ACT CGA GTT CTG ACT ATG AAC AAC CTG C-3' |
| Sequence-based reagent | shHSF1-2 | This paper, from Microsynth |
HSF1 shRNA CDS forward | 5'-CCG GCC AGC AAC AGA AAG TCG TCA ACT CGA GTT GAC GAC TTT CTG TTG CTG GTT TTT G-3' |
| Sequence-based reagent | shHSF1-2 | This paper, from Microsynth |
HSF1 shRNA CDS reverse | 5'-AAT TCA AAA ACC AGC AAC AGA AAG TCG TCA ACT CGA GTT GAC GAC TTT CTG TTG CTG G-3' |
| Chemical compound | L-azetidine-2-carboxylic acid (AZC) | Sigma-Aldrich | #P8783 | |
| Chemical compound | Tunicamycin | Cell Signalling | #12819 | |
| Chemical compound | Propidium iodide (PI) | Cayman Chemical | #14289–10 | |
| Chemical compound | Annexin V | Biolegend | #640906 | |
| Chemical compound | Phalloidin-Alexa488 | Thermo Fisher Scientific | #A12379 | |
| Chemical compound | Diamidino-2-phenylindole dye (DAPI) | Thermo Fisher Scientific | #62248 | 1:30,000 in PBS, from 1 mg/ml stock solution |
| Chemical compound, drug | Abemaciclib | MedChemExpress | #HY-16297A-5MG | |
| Chemical compound, drug | Rapamycin | Sigma-Aldrich | #553210 | |
| Commercial assay, kit | Dual-Luciferase detection kit | Promega | #E1910 | |
| Commercial assay, kit | Alexa Fluor 488 NHS Ester (succinimidyl ester) | Thermo Fisher Scientific | #A20100 | |
| Commercial assay, kit | Click-iT Plus OPP Alexa Fluor 594 | Thermo Fisher Scientific | #C10457 | |
| Commercial assay, kit | N-succinyl-Leu-Leu-Val-Tyr-7-amino-4-methyl-coumarin (suc-LLVY-AMC) | Enzo Life Sciences | #BML-P802-0005 | |
| Commercial assay, kit | CellEvent Senescence Green Flow Cytometry Assay Kit | Invitrogen | #C10840 | |
| Antibody | Anti-GAPDH (Mouse monoclonal) | HyTest Ltd. | 5G4 | 1:1000 |
| Antibody | Anti-Hsp25/27 (Mouse monoclonal) | StressMarq | SMC-114 | 1:1000 |
| Antibody | Anti-puromycin (Mouse monoclonal) | Sigma-Aldrich | MABE343 | 1:22000 |
| Antibody | Anti-Lamin B1 (Rabbit polyclonal) | Cell Signaling Technology | 12586 | 1:1000 |
| Antibody | Anti-Hsp40/Hdj1 (Rabbit polyclonal) | Enzo Lifesciences | ADI-SPA-400 | 1:1000 |
| Antibody | Anti-Hsf1 (Rabbit polyclonal) | Enzo Lifesciences | ADI-SPA-901 | 1:1000 |
| Antibody | Anti-Hsp70 (Mouse monoclonal) | StressMarq | SMC-100 | 1:1000 |
| Antibody | Anti-Phospho-eIF2α (Ser51) (Rabbit polyclonal) | Cell Signaling Technology | 3597 | 1:1000 |
| Antibody | Anti-eIF2α (Rabbit polyclonal) | Cell Signaling Technology | 9722 | 1:1000 |
| Antibody | Anti-Hsp90α (9D2) (Rat monoclonal) | Enzo Lifesciences | ADI-SPA-840 | 1:1000 |
| Antibody | Anti-Hsp90β (scFv H90-10) (Mouse monoclonal) | Geneva Antibody Facility | ABCD_A0870 | 1:2000 |
| Antibody | Anti-mTOR (Rabbit polyclonal) | Cell Signaling Technology | 2983 | 1:2000 |
| Antibody | Anti-Phospho-mTOR (Ser2448) (Rabbit polyclonal) | Cell Signaling Technology | 2971 | 1:1000 |
| Antibody | Anti-Phospho-S6 Ribosomal Protein (Ser235/236) (Rabbit polyclonal) | Cell Signaling Technology | 4858 | 1:1000 |
| Antibody | Anti-S6 Ribosomal Protein (Rabbit polyclonal) | GeneTex | GTX130450 | 1:1000 |
| Antibody | Anti-Hsc70 (Mouse monoclonal) | StressMarq | SMC-151 | 1:2000 |