| Genetic reagent (Mus musculus) |
C57BL/6 N wild-type |
Leibniz Research Institute for Molecular Pharmacology |
RRID:IMSR_JAX:000664
|
|
| Genetic reagent (Mus musculus) |
mDia1 KO |
Bartolini Lab, Columbia University |
Peng et al., 2003; https://doi.org/10.1016/s0960-9822(03)00170–2 |
Genetic knockout of Diaph1 |
| Cell line (Homo sapiens) |
HEK293T |
American Type Culture Collection |
Cat# CRL-3216; RRID:CVCL_0063
|
|
| Antibody |
β-Actin (mouse monoclonal) |
Sigma-Aldrich |
Cat# A5441; RRID:AB_476744
|
IB (1:5000) |
| Antibody |
Bassoon (guinea pig monoclonal) |
Synaptic Systems |
Cat# 141 318; RRID:AB_2927388
|
IC (1:100) |
| Antibody |
CDC42 (rabbit polyclonal) |
Abcam |
Cat# ab64533; RRID:AB_1310067
|
IB (1:1000); IC (1:100) |
| Antibody |
Dynamin1 (rabbit polyclonal) |
Pietro D. Camilli |
Shupliakov et al., 2002; DOI: 10.1126/science.276.5310.29 |
IB (1:2000) |
| Antibody |
GFP (mouse monoclonal) |
Thermo Fisher Scientific |
Cat# A-11120; RRID:AB_221568
|
IC (1:2500) |
| Antibody |
Homer1 (mouse monoclonal) |
Synaptic Systems |
Cat# 160 011; RRID:AB_2120992
|
IC (1:200) |
| Antibody |
Homer1 (rabbit polyclonal) |
Synaptic Systems |
Cat# 160 003; RRID:AB_887730
|
IC (1:200) |
| Antibody |
LAMP1 (rabbit monoclonal) |
Cell Signaling Technology |
Cat# 9091; RRID:AB_2687579
|
IB (1:1000) |
| Antibody |
mDia1 (mouse monoclonal) |
BD Biosciences |
Cat# 610848; RRID:AB_398167
|
IB (1:500); IP (2 µg) |
| Antibody |
mDia1 (rabbit monoclonal) |
Abcam |
Cat# ab129167; RRID:AB_11143749
|
IC (1:100) |
| Antibody |
mDia3 (rabbit polyclonal) |
Sigma-Aldrich |
Cat# HPA005647; RRID:AB_1078657
|
IB (1:1000) |
| Antibody |
Myosin IIb (mouse monoclonal) |
Abcam |
Cat# ab684; RRID:AB_305661
|
IC (1:100) |
| Antibody |
Myosin IIb (rabbit polyclonal) |
Cell Signaling Technology |
Cat# 3404; RRID:AB_126421
|
IB (1:2000) |
| Antibody |
Rac1 (mouse monoclonal) |
BD Biosciences |
Cat# 610650; RRID:AB_397977
|
IB (1:1000); IC (1:50) |
| Antibody |
RFP (rabbit polyclonal) |
Takara Bio |
Cat# 632496; RRID:AB_10013483
|
IC (1:500) |
| Antibody |
RhoA (rabbit monoclonal) |
Cell Signaling Technology |
Cat# 2117; RRID:AB_10693922
|
IB (1:1000); IC (1:100) |
| Antibody |
α-Tubulin (mouse monoclonal) |
Sigma-Aldrich |
Cat# T9026; RRID:AB_47759
|
IB (1:5000) |
| Antibody |
IgG control (mouse monoclonal) |
Thermo Fisher Scientific |
Cat# 31903; RRID:AB_10959891
|
IP (2 μg) |
| Antibody |
vGAT-CypHer5E (rabbit polyclonal) |
Synaptic Systems |
Cat# 131 103CpH; RRID:AB_2189809
|
Live-imaging (1:500) |
| Antibody |
Anti-mouse IgG Atto542 (donkey polyclonal) |
Martin Lehmann |
Gonschior et al., 2022; https://doi.org/10.1038/s41467-022-32533-4
|
IC (1:400) |
| Antibody |
Anti-rabbit IgG Atto542 (donkey polyclonal) |
Martin Lehmann |
Gonschior et al., 2022; https://doi.org/10.1038/s41467-022-32533-4
|
IC (1:400) |
| Antibody |
Anti-mouse Alexa Fluor 594 (goat polyclonal) |
Thermo Fisher Scientific |
Cat# A-11032; RRID:AB_2534091
|
IC (1:200) |
| Antibody |
Anti-rabbit Alexa Fluor 594 (goat polyclonal) |
Thermo Fisher Scientific |
Cat# A-11037; RRID:AB_2534095
|
IC (1:200) |
| Antibody |
Anti-guinea pig Atto647N (camelid monoclonal) |
Synaptic Systems |
Cat# N0602-At647N-S; RRID:AB_2744576
|
IC (1:200) |
| Antibody |
Anti-mouse Alexa Fluor 647 (goat polyclonal) |
Thermo Fisher Scientific |
Cat# A-21235; RRID:AB_2535804
|
IC (1:200) |
| Antibody |
Anti-mouse IRDye 680RD (goat polyclonal) |
LI-COR Biosciences |
Cat# 925–68070; RRID:AB_2651128
|
IB (1:10000) |
| Antibody |
Anti-rabbit IRDye 680RD (goat polyclonal) |
LI-COR Biosciences |
Cat# 926–68071; RRID:AB_10956166
|
IB (1:10000) |
| Antibody |
Anti-mouse IgG, IRDye 800CW (goat polyclonal) |
LI-COR Biosciences |
Cat# 926–32210; RRID:AB_621842
|
IB (1:10000) |
| Antibody |
Anti-rabbit IgG, IRDye 800CW (goat polyclonal) |
LI-COR Biosciences |
Cat# 926–32211; RRID:AB_621843
|
IB (1:10000) |
| Chemical compound, drug |
Phalloidin-Alexa Fluor 594 |
AAT Bioquest |
Cat# ABD-23158 |
IC (1:1000) |
| Recombinant DNA reagent |
Synaptophysin-pHluorin |
Leon Lagnado |
Gonschior et al., 2022; https://doi.org/10.1038/s41467-022-32533-4
|
Expresses rat Synaptophysin-2xpHluorin (inserted between Asn183 – Thr184) under a CMV promotor |
| Recombinant DNA reagent |
vGlut1-pHluorin |
Volker Haucke |
Bolz et al., 2023; https://doi.org/10.1016/j.neuron.2023.08.016
|
Expresses rat vGlut1-pHluorin (inserted between Gly99 – Gly100) under a hSyn1 promotor; lentiviral plasmid |
| Recombinant DNA reagent |
mCherry |
Clontech |
Cat# 632523 |
Expresses mCherry under a CMV promotor |
| Recombinant DNA reagent |
mDia1-WT-mCherry |
This study |
- |
Expresses mouse mDia1-WT-mCherry under a CMV promotor |
| Recombinant DNA reagent |
mDia1-ΔN -mCherry |
This study |
- |
Expresses truncation (first 60 AA) variant of mouse mDia1-mCherry under a CMV promotor |
| Recombinant DNA reagent |
mDia1-WT-SNAP |
This study |
- |
Expresses mouse mDia1-WT-SNAP under a hSyn1 promotor; lentiviral plasmid |
| Recombinant DNA reagent |
mDia1-K994A-SNAP |
This study |
- |
Expresses K994A variant of mDia1-SNAP under a hSyn1 promotor; lentiviral plasmid |
| Recombinant DNA reagent |
Dynamin1-WT-SNAP |
This study |
- |
Expresses mouse Dynamin1-WT-SNAP under a hSyn1 promotor; lentiviral plasmid |
| Recombinant DNA reagent |
Dynamin1-K44A-SNAP |
This study |
- |
Expresses K44A variant of mouse Dynamin1-SNAP under a hSyn1 promotor; lentiviral plasmid |
| Recombinant DNA reagent |
vGlut1-mCherry |
Franck Polleux |
Kwon et al., 2016; https://doi.org/10.1371/journal.pbio.1002516
|
Expresses rat vGlut1-mCherry under a CAG promotor |
| Recombinant DNA reagent |
Rac1-CA |
Addgene |
Cat# 12983; RRID:Addgene_12983
|
Expresses Q61L variant of human myc-Rac1 under a CMV promotor |
| Recombinant DNA reagent |
Rac1-DN |
Addgene |
Cat# 12984; RRID:Addgene_12984
|
Expresses T17N variant of human myc-Rac1 under a CMV promotor |
| Recombinant DNA reagent |
RhoA-WT |
Theofilos Papadopoulos |
Reddy-Alla et al., 2010; https://doi.org/10.1111/j.1460-9568.2010.07149.x
|
Expresses human 3xHA-RhoA-WT under a CMV promotor |
| Recombinant DNA reagent |
RhoA-DN |
Theofilos Papadopoulos |
Reddy-Alla et al., 2010; https://doi.org/10.1111/j.1460-9568.2010.07149.x
|
Expresses T19N variant of human 3xHA-RhoA under a CMV promotor |
| Recombinant DNA reagent |
RhoB-WT |
Harry Mellor |
Mellor, 1998; https://doi.org/10.1074/jbc.273.9.4811
|
Expresses human myc-RhoB-WT under a CMV promotor |
| Recombinant DNA reagent |
RhoB-DN |
Harry Mellor |
Mellor, 1998; https://doi.org/10.1074/jbc.273.9.4811
|
Expresses T19N variant of human myc-RhoB under a CMV promotor |
| Recombinant DNA reagent |
shCTR (transfected) |
Sigma-Aldrich |
Cat# SHC016 |
No murine targets |
| Recombinant DNA reagent |
shmDia1 (transfected) |
Sigma-Aldrich |
Cat# TRCN0000108685 |
Targets 3'UTR of NM_007858
|
| Recombinant DNA reagent |
shRhoA (transfected) |
Sigma-Aldrich |
Cat# TRCN0000302388 |
Targets CDS of NM_016802
|
| Recombinant DNA reagent |
shRhoB (transfected) |
Sigma-Aldrich |
Cat# TRCN0000294874 |
Targets CDS of NM_007483
|
| Recombinant DNA reagent |
shCTRmiR |
This study |
- |
Expresses rat Synaptophysin-2xpHluorin under a CMV promotor and shRNA embedded into a microRNA (shRNAmiR) with no murine targets |
| Recombinant DNA reagent |
shmDia1miR |
This study |
- |
Expresses rat Synaptophysin-2xpHluorin under a CMV promotor and shRNAmiR against the CDS of mouse Diaph1 (mDia1) |
| Recombinant DNA reagent |
shCTR (transduced) |
Christian Rosenmund |
Watanabe et al., 2014; https://doi.org/10.1038/nature13846
|
Expresses NLS-RFP or BFP under a hSyn1 promotor and shRNA against no murine target (msClathrin scrambled) under a U6 promotor |
| Recombinant DNA reagent |
shmDia1 (transduced) |
This study |
- |
Expresses NLS-RFP or BFP under a hSyn1 promotor and shRNA against the 3'UTR of mouse Diaph1 (mDia1) under a U6 promotor |
| Recombinant DNA reagent |
shmDia3 (transduced) |
This study |
- |
Expresses NLS-RFP or BFP under a hSyn1 promotor and shRNA against the CDS of mouse Diaph2 (mDia3) under a U6 promotor |
| Recombinant DNA reagent |
MD2.G |
Addgene |
Cat# 12259; RRID:Addgene_12259
|
Expresses lentiviral VSV-G envelope protein |
| Recombinant DNA reagent |
psPAX2 |
Addgene |
Cat# 12260; RRID:Addgene_12260
|
Expresses lentiviral packaging protein |
| Recombinant DNA reagent |
pOrange GFP-β-Actin KI |
Addgene |
Cat#131479; RRID:Addgene_131479
|
gRNA and GFP donor for endogenous N-terminal tagging of β-Actin (amino acid position: D2) by targeting Actb |
| Chemical compound, drug |
Dynasore |
Sigma-Aldrich |
Cat# D7693 |
80 μM 10 min |
| Chemical compound, drug |
EHT 1864 |
MedChemExpress |
Cat# HY-16659 |
10 μM acute (CypHer); 2 h (IC/EM) |
| Chemical compound, drug |
IMM-01 |
Sigma-Aldrich |
Cat# SML1064 |
10 μM acute |
| Chemical compound, drug |
Jasplakinolide |
Sigma-Aldrich |
Cat# J4580 |
8 μM acute (JLY); 1 μM 30 min |
| Chemical compound, drug |
Latrunculin A |
Cayman Chemical |
Cat# CAY10684 |
5 µM acute (JLY) |
| Chemical compound, drug |
ML141 |
MedChemExpress |
Cat# HY-12755 |
10 µM acute |
| Chemical compound, drug |
Rhosin |
MedChemExpress |
Cat# HY-12646 |
10 µM 2 h |
| Chemical compound, drug |
Tetrodotoxin |
Carl Roth |
Cat# 6973.1 |
1 µM 36 h |
| Chemical compound, drug |
Y-27632 |
Tocris |
Cat# 1254 |
1 µM acute (JLY) |
| Commercial assay or kit |
Rhotekin-Rho binding domain (RBD) beads |
Cytoskeleton Inc |
Cat# RT02 |
60 μg |
| Commercial assay or kit |
PAK-p21 binding domain (PBD) beads |
Cytoskeleton Inc |
Cat# PAK02 |
20 μg |
| Commercial assay or kit |
ProFection Mammalian Transfection System – Calcium Phosphate |
Promega |
Cat# E1200 |
|
| Commercial assay or kit |
Gibson Master Mix |
New England Biolabs Inc |
Cat# E2611L |
|
| Commercial assay or kit |
Q5 site-directed mutagenesis kit |
New England Biolabs Inc |
Cat# E0552S |
|
| Software, algorithm |
Fiji |
NIH |
RRID:SCR_002285
|
|
| Software, algorithm |
Prism |
GraphPad |
RRID:SCR_002798
|
Version 9.5.1. |
| Software, algorithm |
Image Lab Software |
Bio-Rad |
RRID:SCR_014210
|
Version 6.0.1 |
| Software, algorithm |
Image Studio Lite |
LI-COR Biosciences |
RRID:SCR_013715
|
Version 5.2.5 |
| Software, algorithm |
pHluorin ROI sector |
Github |
https://github.com/DennisVoll/pHluorin_ROI_selector/; Voll, 2020
|
|
| Software, algorithm |
SynActJ |
Martin Lehmann |
Schmied et al., 2021; https://doi.org/10.3389/fcomp.2021.777837
|
|
| Software, algorithm |
Macro_plot_lineprofile_multicolor |
Kees Straatman |
Gerth et al., 2017; https://doi.org/10.1016/j.str.2019.03.020
|
|
| Software, algorithm |
MaxQuant |
Jürgen Cox |
https://www.maxquant.org/maxquant/
|
Version 1.6.1.0 |
| Software, algorithm |
Perseus |
Jürgen Cox |
https://www.maxquant.org/perseus/
|
Version 1.6.7.0 |
| Sequence-based reagent |
see Supplementary file 1
|
|
|
|
