KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-mNeonGreen nAb | Allele Biotechnology | Cat# ABP-NAB-MNGAB; RRID: AB_2629489 |
| Rabbit anti-HaloTag pAb | Promega | Cat# G9281; RRID:AB_713650 |
| Bacterial and Virus Strains | ||
| N/A | ||
| Biological Samples | ||
| N/A | ||
| Chemicals, Peptides, and Recombinant Proteins | ||
| EZ-Link NHS-PEG4-biotin | ThermoFisher | Cat# 21330 |
| mPEG-Silane, MW 5,000 | Laysan Bio, Arab, AL | Cat# MPEG-SIL-5000 |
| Biotin-PEG-Silane, MW 5,000 | Laysan Bio, Arab, AL | Cat# Biotin-PEG-SIL-5K |
| 2-[methoxy(polyethylenxy)9–12-propyl] trimethoxysilane | Gelest, Morrisville, PA | Cat# SIM6492.72 |
| Neutravidin | ThermoFisher | Cat# 31000 |
| JF646 HaloTag ligand | Laboratory of Luke D. Lavis | |
| Critical Commercial Assays | ||
| N/A | ||
| Deposited Data | ||
| N/A | ||
| Experimental Models: Cell Lines | ||
| N/A | ||
| Experimental Models: Organisms/Strains | ||
| C. elegans wild-type strain | Caenorhabditis Genetics Center | N2 |
|
C. elegans unc-119 injection
strain Genotype: unc-119(ed3) III |
Caenorhabditis Genetics Center | HT1593 |
|
C. elegans transgenic strain
expressing mNG::HaloTag Genotype: cpIs90[Pmex-5::mNG::HaloTag::tbb-2 3′UTR + LoxP] II |
This study | LP539 |
|
C. elegans transgenic strain
expressing mNG::mKate2 Genotype: cpIs71[Pmex-5::mNG::mKate2::tbb-2 3′UTR + SEC] II |
This Study | LP420 |
|
C. elegans transgenic strain
expressing mNG::mKate2 Genotype: cpIs72[Pmex-5::mNG::mKate2::tbb-2 3′UTR + SEC] II |
This Study | LP421 |
|
C. elegans transgenic strain
expressing mSc::HaloTag Genotype: cpIs132[Pmex-5::mScarlet-I::HaloTag::tbb-2 3′UTR + LoxP] II |
This Study | LP734 |
|
C. elegans transgenic strain
expressing mNG::AraD Genotype: cpIs36[Pmex-5::mNG::3xFlag::AraD::tbb-2 3′UTR + unc-119(+)] II; unc-119(ed3) III |
This Study | LP230 |
|
C. elegans strain with
endogenously tagged mNG::aPKC Genotype: pkc-3(cp41[mNG:3xFlag::pkc-3]) II |
This Study | LP212 |
|
C. elegans strain with
endogenously tagged PAR-6::HaloTag Genotype: par-6(cp346[PAR-6::HaloTag]) I |
This Study | LP654 |
|
C. elegans strain with
endogenously tagged mNG::aPKC and PAR-6::HaloTag Genotype: par-6(cp346[PAR-6::HaloTag]) I; pkc-3(cp41[mNeonGreen:3xFlag::pkc-3]) II |
This Study | LP747 |
|
C. elegans strain with
endogenously tagged PAR-6::mNG Genotype: par-6(cp45[par-6::mNG::3xFlag + LoxP unc-119(+) LoxP]) I; unc-119(ed3) III |
This Study | LP216 |
|
C. elegans strain with
endogenously tagged mNG::PAR-3 Genotype: par-3(cp54[mNG::3xFlag::par-3]) III |
This Study | LP242 |
|
C. elegans strain with
endogenously tagged PAR-6::mKate2 Genotype: par-6(cp60[par-6::mKate2::3xMyc + LoxP unc-119(+) LoxP]) I; unc-119(ed3) III |
This Study | LP244 |
|
C. elegans strain with
endogenously tagged mNG::PAR-3 and PAR-6::mKate2 Genotype: par-6(cp60[par-6::mKate2::3xMyc + LoxP unc-119(+) LoxP]) I; par-3(cp54[mNG::3xFlag::par-3]) III |
This Study | LP282 |
|
C. elegans strain with
endogenously tagged mNG::PAR-3 and PAR-6::HaloTag Genotype: par-6(cp346[PAR-6::HaloTag]) I; par-3(cp54[mNeonGreen::3xFlag::par-3]) III |
This Study | LP749 |
|
C. elegans strain with RRKEEE
mutations and an N-terminal mNG tag in endogenous
par-3 Genotype: par-3(cp344[mNG^3xFlag::PAR-3 R69E R100E K103E]) III |
This Study | LP651 |
|
C. elegans strain with TTDE
mutations and an N-terminal mNG tag in endogenous
par-3 Genotype: par-3(cp327[mNG^3xFlag::PAR-3 T67D T71E]) III |
This Study | LP625 |
|
C. elegans strain with
endogenously tagged mNG::PAR-2 Genotype: par-2(cp329[mNG^PAR-2]) III |
This Study | LP637 |
|
C. elegans strain with
endogenously tagged mNG::PAR-2 and PAR-6::mKate2 Genotype: par-6(cp60[par-6::mKate2::3xMyc + LoxP unc-119(+) LoxP]) I; par-2(cp329[mNG^PAR-2]) III |
This Study | LP675 |
|
C. elegans strain with RRKEEE
mutations and an N-terminal HaloTag in endogenous
par-3 Genotype: par-3(cp354[HaloTag^PAR-3 R69E R100E K103E]) III |
This Study | LP676 |
|
C. elegans strain with RRKEEE
mutations and an N-terminal HaloTag in endogenous
par-3, plus endogenously-tagged mNG::PAR-2 and
PAR-6::mKate2 Genotype: par-6(cp60[par-6::mKate2::3xMyc + LoxP unc-119(+) LoxP]) I; par-2(cp329[mNG^PAR-2]) par-3(cp355[HaloTag^PAR-3 R69E R100E K103E]) III |
This Study | LP677 |
|
C. elegans strain with
endogenously-tagged mNG::MEX-5 Genotype: mex-5(cp119[mNG^3xFlag::mex-5]) IV |
Dickinson et al. 2015 | LP367 |
|
C. elegans strain with RRKEEE
mutations and an N-terminal HaloTag in endogenous
par-3, plus endogenously-tagged
mNG::MEX-5 Genotype: par-3(cp354[HaloTag^PAR-3 R69E R100E K103E]) III; mex-5(cp119[mNG^3xFlag::mex-5]) IV |
This Study | LP679 |
|
C. elegans transgenic strain
expressing GFP::PGL-1 Genotype: bnIs1[pie-1::GFP::pgl-1 + unc-119(+)] I |
Cheeks et al. 2004 | SS747 |
|
C. elegans strain with RRKEEE
mutations and an N-terminal HaloTag in endogenous
par-3, also expressing GFP::PGL-1 from a
transgene Genotype: bnIs1[pie-1::GFP::pgl-1 + unc-119(+)] I; par-3(cp354[HaloTag^PAR-3 R69E R100E K103E]) III |
This Study | LP680 |
|
C. elegans strain with
endogenously tagged NMY-2::mKate2 Genotype: nmy-2(cp52[nmy-2::mkate2 + LoxP unc-119(+) LoxP]) I; unc-119(ed3) III |
This Study | LP229 |
|
C. elegans strain with
endogenously tagged mNG::PAR-3 and NMY-2::mKate2 Genotype: nmy-2(cp52[nmy-2::mkate2 + LoxP unc-119(+) LoxP]) I; par-3(cp54[mNeonGreen::3xFlag::par-3]) III |
This Study | LP636 |
|
C. elegans strain with
phosphomimetic mutations in two putative PLK-1 phosphorylation sites,
plus an N-terminal mNG tag, in endogenous
par-3 Genotype: par-3(cp357[mNG^3xFlag::PAR-3 T32E T89E]) III |
This Study | LP681 |
|
C. elegans strain with
phosphomimetic mutations in two putative PLK-1 phosphorylation sites,
plus an N-terminal mNG tag, in endogenous
par-3 Genotype: par-3(cp358[mNG^3xFlag::PAR-3 T32E T89E]) III |
This Study | LP682 |
| Oligonucleotides | ||
| N/A | ||
| Recombinant DNA | ||
| N/A | ||
| Software and Algorithms | ||
| SiMPull analysis software for MATLAB | This paper | https://github.com/dannyhmg/SiMPull-Analysis-Software |
| Probabilistic Segmentation | Laboratory of Paul Maddox (Padeganeh et al. 2013) | |
| Bayesian step detection | Laboratory of Vijay Pande (Ensign and Pande 2010) | |
| Ezyfit toolbox for MATLAB | MATLAB File Exchange | Version 2.44; http://www.mathworks.com/matlabcentral/fileexchange/10176-ezyfit-2-44 |
| U-track particle tracking software for MATLAB | Jaqaman et al. 2008 | Version 2.1.3; http://lccb.hms.harvard.edu/software.html |
| MSDanalyzer for MATLAB | Tarantino et al. 2014 | https://tinevez.github.io/msdanalyzer/ |
| PIVlab | Thielicke and Stamhuis, 2014 | Version 1.4; http://pivlab.blogspot.com/ |
| Other | ||
| N/A | ||