| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit polyclonal anti-Par3 | Merck | 07-330; RRID:AB_2101325 |
| Mouse monoclonal anti-aPKC | Santa Cruz | sc-17837; RRID:AB_2172068 |
| Mouse polyclonal anti-α K40 acetylated tubulin | HPA cultures | C3B9 |
| Mouse monoclonal anti-PC | Roche | HPC4 |
| Rabbit polyclonal anti-GST | Abcam | ab19256; RRID:AB_444809 |
| Rat monoclonal anti-HA | Roche | 3F10; RRID:AB_2314622 |
| Goat polyclonal anti-rabbit Alexa-555 | Thermo | A32732; RRID:AB_2633281 |
| Goat polyclonal anti-rabbit Alexa-647 | Thermo | A21246; RRID:AB_2535814 |
| Goat polyclonal anti-mouse Alexa-647 | Thermo | A-21235; RRID: AB_2535804 |
| Donkey polyclonal anti-rat Cy3 | Jackson ImmunoResearch | AB_2340667 |
| Rabbit polyclonal anti-Camsap2 | Proteinbiotech | 17880-1-AP; RRID:AB_2068826 |
| rabbit polyclonal anti-Kif2A | Novus Biologicals | NB500-180; RRID:AB_10002348 |
| Chemicals, peptides, and recombinant proteins | ||
| DMEM + Glutamax | Gibco | 10566016 |
| Penicillin-Streptomycin | Gibco | 15140122 |
| DMEM | ThermoFisher Scientific | 41966 |
| MEM | ThermoFisher Scientific | 11140035 |
| DMEM F12 | ThermoFisher Scientific | 21041025 |
| Glutamax | ThermoFisher Scientific | 35050061 |
| sodium pyruvate | ThermoFisher Scientific | 11360070 |
| Neurobasal A | ThermoFisher Scientific | 12348017 |
| B27 | ThermoFisher Scientific | 10889-038 |
| β -mercaptoethanol | ThermoFisher Scientific | 31350-010 |
| Hygromycin B Gold | InvivoGen | Ant-hg-1 |
| puromycin | ThermoFisher Scientific | A11138-03 |
| Foetal Bovine Serum | Gibco | 10270106 |
| Donor Bovine Serum | Gibco | 16030074 |
| Lipofectamine 3000 | Invitrogen | L3000001 |
| MEK inhibitor | Cambridge Stem Cell Institute | PD0325901 |
| GSK3 inhibitor | Cambridge Stem Cell Institute | CHIR99021 |
| Leukaemia Inhibitory Factor | Marko Hyvonen’s lab | https://hyvonen.bioc.cam.ac.uk/proteins/mLIF_datasheet.pdf |
| MycoAlert Mycoplasma Detection kit | Lonza | LT07-118 |
| Schneider Medium | ThermoFisher Scientific | 21720024 |
| Insect Express medium | Lonza | BELN12-730Q |
| Effectene | Qiagen | 301425 |
| Pluronic F-68 | Gibco | 24040032 |
| A(d) | Ben-Sasson et al.20 | N/A |
| A(d)-mCherry | Ben-Sasson et al.20 | N/A |
| B(c)-GFP | Ben-Sasson et al.20 | N/A |
| B(c)-SpyCatcher | This study | N/A |
| SpyTag-DLL4 | Watson et al.48 | N/A |
| Fibronectin | Sigma | F1141 |
| GFP protein | This study | N/A |
| Fipper-TR | Spirochrome | SC020 |
| Concanavalin A | Sigma | L7647-25MG |
| Experimental model: Cell lines | ||
| NiH/3T3 Flp-In cells (3T3 cells) | Invitrogen | R76107 |
| Stable 3T3 cells expressing GG-GFP∗ | This study | N/A |
| Stable 3T3 cells expressing GG-Par3(180kD)∗ | This study | N/A |
| Stable 3T3 cells expressing GG-Par3(100kD)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(83-1319)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(S824A/S826A)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(S824D/S826D)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(100kD, Δ1-83)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(100kD Δ1-83 Δ728-813)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par3(100kD G600A/G602A Δ1-83 Δ728-813)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6A∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6B∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6G∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6A(K19A)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6A(1-121)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6A(121-346)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- Par6A(247-346)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- PAR6A(121-346, LGF169-171AAA)∗ | This study | N/A |
| Stable 3T3 cells expressing GG-Jo-Par6A-In∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(E85A/R91A)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(K274W)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(A129E)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(E85A/R91A/K274W)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(E85A/R91A/A129E)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- aPKCι(Δ857-862)∗ | This study | N/A |
| Stable 3T3 cells expressing GG- mInsc∗ | This study | N/A |
| Stable 3T3 cells expressing GG- LGN∗ | This study | N/A |
| Stable 3T3 cells expressing His-PC-GBP-TM-VSVG-Par6A | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-mScarlet | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-aPKCι | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-Par3(180kD) | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-Par3(100kD) | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-mInsc | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-Par3(S824A/S826A) | This study | N/A |
| Stable 3T3 cells expressing TetOn∗∗ His-PC-GBP-TM-VSVG-Par3(S824D/S826D) | This study | N/A |
| E14 WT Mouse ESCs | Jenny Nichols | N/A |
| Drosophila S2 Cells | ThermoFisher Scientific | R69007 |
| U2OS | ATCC | HTB-96 |
| Stable U2OS cells expressing FLAG-Notch1-EGFP | Malecki et al.49 | N/A |
| Experimental model: Drosophila Melanogaster stocks | ||
| w1118 | Bloomington | #3605 |
| Neur>Gal4 | Bellaïche et al.50 | N/A |
| UAS>mRFP-PonlD | Emery et al.51 | N/A |
| Baz-mScarlet knock-in | Houssin et al.52 | N/A |
| Jupiter-GFP knock-in | Bloomington | #6836 |
| Software and algorithms | ||
| Fiji | Schindelin et al.53 | https://imagej.net/Fiji |
| MATLAB 2020b | Mathworks | https://uk.mathworks.com/products/matlab.html |
| Adobe Premiere 27.3.1 | ADOBE | https://www.adobe.com/ |
| GPU Drift correction | Planelles-Herrero et al.10 | https://github.com/deriverylab / GPU_registration |
| Wavelet “à trous” GPU denoising | Planelles-Herrero et al.10 | https://github.com/deriverylab / GPU_wavelet_a_trous |
| 3D array colocalization | This study | https://github.com/deriverylab/3D_array_colocalization; https://doi.org/10.5281/zenodo.8357466 |
| CudaDecon | Janelia Research Campus | https://github.com/scopetools/cudaDecon |
| Lsfmtools | James Manton and Jérôme Boulanger | https://github.com/jdmanton/lsfm_tools |
| LimeSeg | Machado et al.54 | https://imagej.net/plugins/limeseg |
| SymPhoTime 64 2.0 | PicoQuant | https://www.picoquant.com |
| Metamorph 7.10.1.161 | Molecular devices | www.moleculardevices.com |
| Napari | Ahlers et al.55 | https://napari.org/ |
| Protein structure predictions | ||
| Par6A | AlphaFold2 | https://alphafold.ebi.ac.uk/entry/Q9NPB6 |
∗ GG refers to the tricistronic His-PC-GBP-TM-VSVG-GBP + Flag iRFP670-Jupiter + PC-GFP-[protein of interest]
∗∗ TetOn refers to a bicistronic rtTA3 + Tet promoter driving the given ORF