Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | BL21 (DE3) Codon-Plus RIL | Agilent | 230245 | Strain for expressing recombinant proteins |
| Strain, strain background (Escherichia coli) | XL10-Gold ultracompetent cells | Agilent | 200314 | Strain used for molecular biology and creating recombinant DNA |
| Cell line (Homo-sapiens) | Dvl TKO HEK293T | Stephane Angers, University of Toronto See Gammons et al., 2016b https://doi.org/10.1242/jcs.195685 |
CRISPR/Cas9 deletion of hDvl1, hDvl2, and hDvl3 Authentication methods: deletions were confirmed by genomic DNA sequencing; immunoblotting with anti-Dvl2 (Figure 7—figure supplement 1) Mycoplasma contamination testing status: Tested |
|
| Cell line (Homo-sapiens) | Dvl2-GFP/dsRed-Axin1 HEK293T | This paper; cells were derived from cells purchased from the European Collection of Cell Cultures | C-terminal tagging by CRISPR/Cas9 gene editing Authentication methods: STR DNA profiling; flow cytometry and fluorescence microscopy using GFP/RFP; local genomic DNA sequencing; and anti-Dvl2 (Figure 7—figure supplement 1) and anti-Axin1 immunoblots Mycoplasma contamination testing status: Tested Contact Bienz laboratory for distribution |
|
| Cell line (mouse) | L and L3A | ATCC | L CRL-2648 L3A CRL-2647 |
Used for generating control and Wnt-conditioned media; See https://web.stanford.edu/group/nusselab/cgi-bin/wnt/ |
| Recombinant DNA reagent | pCS2+ | This paper | Ampicillin resistance; expression in mammalian cell culture; includes a N-terminal M2 Flag tag Contact Weis lab for distribution |
|
| Recombinant DNA reagent | pCDF-Duet-His6-MBP-TEV | Novagen; modified in Weis lab to include MBP-TEV | 71340 | Streptomycin resistance; expression in bacterial cultures Contact Weis lab for distribution |
| RecombinantDNA reagent | pGEX-TEV |
Choi et al., 2006
https://doi.org/10.1074/jbc.M511338200 |
Ampiicillin resistance; expression in bacterial cultures; pGEX-KG plasmid (ATCC) with a new TEV protease site Contact Weis lab for distribution |
|
| Chemical compound, drug | Fetal Bovine Serum | Gemini GemCell, U.S. Origin | Used to generating control and Wnt-conditioned media; provides low basal activity | |
| Chemical compound, drug | LB Broth, Miller, granules | Fisher BioReagents | ||
| Chemical compound, drug | Amylose agarose | New England BioLabs | For purification by MBP affinity | |
| Chemical compound, drug | Glutathione agarose | Pierce | For purification by GST affinity | |
| Chemical compound, drug | Negative stain grids (carbon-coated copper) | EMS | CF200-Cu | |
| Chemical compound, drug | Cryo-EM lacey grids | EMS | LC200-Cu | Freezing done with Leica EM GP |
| Antibody | Dvl2 polyclonal antibody | Cell Signaling Technology | 3216 | IB (1:1000) RRID:AB_2093338 |
| Antibody | Axin1 antibody | Cell Signaling Technology | C76H11 | IB (1:1000) RRID:AB_2054638 |
| Antibody | Beta-catenin monoclonal antibody L54E2, AlexaFluor647-conjugated | Cell Signaling Technology | 4627 | IF (1:300) RRID:AB_10691326 |
| Recombinant DNA reagent |
Dvl2 (mouse) | This paper | GenBank U24160.2 | residues 2–736; DIX = 12–92; Plasmid in pCS2+-M2-Flag; contact Weis lab for distribution |
| Recombinant DNA reagent |
Axin1 (human) | This paper | NCBI NM_181050.3 | corresponds to residues1–826 of NP_851393.1 DAX = 743–826 DI-DAX = 599–826 Plasmid in pCAN-M2S-myc; contact Weis lab fordistribution |
| Recombinant DNA reagent | Tankyrase2 SAM domain (human) | Nai-Wen Chi (Addgene plasmid # 34691) | NCBI NP_079511.1 | residues 867–940 |
| Recombinant DNA reagent | Sm1 residues 1–77 heptamerization domain (archaea) |
Integrated DNATechnologies/This paper | NCBI WP_010878376.1 | Contact Weis lab for distribution PDB: 1LJO; Törö et al., 2002 |
| Commercial assay or kit | MALS | Wyatt | See main text for more details | |
| Commercial assay or kit | S75, S200, Superose 6 10/300 | Pharmacia/GE | 24 mL ‘increase’ columns are tolerant of high flow rates and have slight differences in elution profile | |
| Commercial assay or kit | TopFlash Dual-Light Reporter Gene Assay System | ThermoFisher/Applied Biosystems | T1005 | |
| Commercial assay or kit | AlexaFluor-488/647 C2 maleimide | ThermoFisher/Molecular Probes | See Materials and methods for more details | |
| Commercial assay or kit | Phusion HiFi DNA polymerase | Fermentas/Thermo Fisher | ||
| Commercial assay or kit | FastDigest Restriction endonucleases | Fermentas/Thermo Fisher | ||
| Commercial assay or kit | Gibson Assembly HiFi 1-Step Kit | SGI DNA | GA1100-10 | |
| Commercial assay or kit | Stain-free TGX | Biorad | Specifically visualizes Trp-containing proteins, using Gel Doc EZ | |
| Commercial assay or kit | Any Kd TGX | Biorad | 4569036 | Used for native gel runs |
| Commercial assay or kit | LiCOR IR-dye secondary antibody and Odyssey 3.0 imaging system | LiCOR, Inc | Used for visualizing immunoblots and quantifying sedimentation assays | |
| Software, algorithm | RELION 3.0.8 |
He and Scheres, 2017
https://doi.org/10.1016/j.jsb.2017.02.003 Scheres, 2012 https://doi.org/10.1016/j.jmb.2011.11.010 Zivanov et al., 2018 https://doi.org/10.7554/eLife.42166 |
RRID:SCR_016274 | |
| Software, algorithm | CTFFIND-4.1 | Rohou and Grigorieff, 2015 https://doi.org/10.1016/j.jsb.2015.08.008 | RRID:SCR_016732 | |
| Software, algorithm | Phenix | Afonine et al., 2018 https://doi.org/10.1107/S2059798318006551 | RRID:SCR_014224 | |
| Software, algorithm | Coot | Emsley et al., 2010 https://doi.org/10.1107/S0907444910007493 | RRID:SCR_014222 | |
| Software, algorithm | FIJI | Schindelin et al., 2012 https://doi.org/10.1038/nmeth.2019 | RRID:SCR_002285 | |
| Software, algorithm | Astra 6 | Wyatt Technologies | RRID:SCR_016255 | |
| Software, algorithm | SBGrid | Morin et al., 2013 https://doi.org/10.7554/eLife.01456 | RRID:SCR_003511 | |
| Software, algorithm | UCSF Chimera | Pettersen et al., 2004 https://doi.org/10.1002/jcc.20084 | RRID:SCR_004097 | v1.14 |
| Software, algorithm | PyMOL | Schrödinger, LLC | RRID:SCR_000305 | v2.3.3 |
| Software, algorithm | GraphPad Prism 8.0.2 | GraphPad Software, Inc | Version 263 RRID:SCR_002798 |
|
| Software, algorithm | u-track | Jaqaman et al., 2008 https://doi.org/10.1038/nmeth.1237 | ||
| Software, algorithm | Matlab | The MathWorks, Inc | Version 9.6.0.1072779 (R2019a) | |
| Software, algorithm | BioRender | BioRender - biorender.com | Used for Figure 7a |