Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Mus musculus) | C57BL/6 | Jackson Laboratories | Cat# 002595; RRID:MGI:5656552 |
|
| Strain, strain background (M. musculus) | M2A-GFP KI | Robert Adelstein, NHLBI/NIH | ||
| Cell line (M. musculus) | A20 | American Type Culture Collection | ATCC TIB-208; RRID:CVCL_1940 |
IgG+ B cell line |
| Recombinant DNA reagent | GFP-F-Tractin | Michael Schell (Uniformed Services University, MD) | ||
| Recombinant DNA reagent | tdTomato-F-Tractin | Michael Schell (Uniformed Services University, MD) | ||
| Chemical compound, drug | Alexa Fluor 488 Phalloidin | Thermo Fisher Scientific | Cat# A12379 | Fluorescence labeling (1:500) |
| Chemical compound, drug | Alexa Fluor 568 Phalloidin | Thermo Fisher Scientific | Cat# A12380 | Fluorescence labeling (1:500) |
| Chemical compound, drug | CellMask Deep Red Plasma Membrane Stain |
Thermo Fisher Scientific | Cat# C10046 | Cell labeling (1:10000) |
| Chemical compound, drug | DMSO | MilliporeSigma | Cat# D4540 | |
| Chemical compound, drug | SMIFH2 | MilliporeSigma | Cat# 344092 | 25 μM |
| Chemical compound, drug | CK-666 | MilliporeSigma | Cat# 182515 | 100 μM |
| Chemical compound, drug | (S)-nitro- blebbistatin (pnBB) |
Cayman Chemicals | Cat# 24171 | 25 μM |
| Chemical compound, drug | Para-amino blebbistatin | Cayman Chemicals | Cat# 22699 | 25 μM |
| Chemical compound, drug | Zombie Yellow viability stain |
BioLegend | Cat# 423103 | Dead cell staining (1:300) |
| Chemical compound, drug | 18:1 Biotinyl Cap PE IN CHLOROFORM 1,2-dioleoyl- sn-glycero-3-phosphoethanolamine- N-(cap biotinyl) |
Avanti Polar Lipids | Cat# 870273C | |
| Chemical compound, drug | 18:1 DGS-NTA(Ni) in Chloroform 1,2-dioleoyl-sn-glycero- 3-[(N-(5-amino-1-carboxypentyl) iminodiacetic acid)succinyl] (nickel salt) |
Avanti Polar Lipids | Cat# 790404C | |
| Chemical compound, drug | 18:1 (9-Cis) PC (DOPC) in CHLOROFORM 1,2-dioleoyl-sn-glycero- 3-phosphocholine |
Avanti Polar Lipids | Cat# 850375C | |
| Chemical compound, drug | TransFluoSpheres (633/720) | Thermo Fisher Scientific | Cat# T8870 | |
| Other | Escherichia coli O111:B4 LPS | MilliporeSigma | Cat# L2630 | Cell culture 2.5 µg/ml |
| Biological sample (Ovis aries) | Sheep’s red blood cells | Innovative Research Novi | Cat# ISHRBC100P15ML | Injection 2 × 108 cells |
| Antibody | Alexa Fluor 488- conjugated goat, anti-rabbit, polyclonal |
Jackson Immuno Research |
Cat# 111-545-003; RRID:AB_2338046 |
Immunofluorescence (1:500) |
| Antibody | Alexa Fluor 594 conjugated goat, anti-rabbit, polyclonal |
Jackson Immuno Research |
Cat# 111-585-003; RRID:AB_2338059 |
Immunofluorescence (1:500) |
| Antibody | Alexa Fluor 647- conjugated goat, anti-rabbit, polyclonal | Jackson Immuno Research |
Cat# 111-605-003; RRID:AB_2338072 |
Immunofluorescence (1:500) |
| Antibody | Goat anti-mouse IgG, Fcγ fragment-specific, polyclonal | Jackson Immuno Research |
Cat# 115-005-008; RRID:AB_2338449 |
Coverslip coating 2.5 µg/cm2 |
| Antibody | Goat anti-mouse IgM, µ-chain-specific, polyclonal | Jackson Immuno Research |
Cat# 115-005-020; RRID:AB_2338450 |
Coverslip coating 2.5 µg/cm2 |
| Antibody | Goat anti-rabbit IgG (H + L) Poly-HRP, polyclonal | Thermo Fisher | Cat# 32260; RRID:AB_1965959 |
Western blot (1:3000) |
| Antibody | Rabbit anti-DIAPH1, polyclonal | Thermo Fisher | Cat# PA5-27607; RRID:AB_2545083 |
Western blot (1:250) |
| Antibody | β-actin antibody (C4), mouse monoclonal | Santa Cruz | Cat# SC-47778 HRP; RRID:AB_2714189 |
Western blot (1:5000) |
| Antibody | Rabbit anti-CD79a, polyclonal | Cell Signaling Technologies | Cat# 3351; RRID:AB_2075745 |
Immunofluorescence (1:250) |
| Antibody | Rabbit anti-phospho-CD79a, polyclonal | Cell Signaling Technologies | Cat# 5173; RRID:AB_10694763 |
Immunofluorescence (1:250) |
| Antibody | Rabbit anti-CD19, polyclonal | Cell Signaling Technologies | Cat# 3574; RRID:AB_2275523 |
Immunofluorescence (1:250) |
| Antibody | Rabbit anti-phospho-CD19, polyclonal | Cell Signaling Technologies | Cat# 3571; RRID:AB_2072836 |
Immunofluorescence (1:250) |
| Antibody | Rabbit anti-M2A, polyclonal | MilliporeSigma | Cat# M8064; RRID:AB_260673 |
Immunofluorescence (1:200) |
| Antibody | TruStain FcX PLUS (anti-mouse CD16/32) antibody, rat monoclonal | BioLegend | Cat# 156604; RRID:AB_2783138 |
FcR block (0.25 μg/106 cells) |
| Antibody | Pacific Blue anti-mouse CD38, rat monoclonal | BioLegend | Cat# 102719; RRID:AB_10613289 |
FACS (1:100) |
| Antibody | PerCP/Cyanine5.5 anti-mouse/human CD45R/B220, rat monoclonal | BioLegend | Cat# 103235; RRID:AB_893356 |
FACS (1:100) |
| Antibody | APC anti-MU/HU GL7 antigen, rat monoclonal | BioLegend | Cat# 144617; RRID:AB_2800674 |
FACS (1:200) |
| Recombinant DNA reagent | mNeonGreen-F-Tractin | This paper | See Materials and methods | |
| Sequence-based reagent | Non-targeting miRNA |
This paper | miRNA | ACCTAAGGTTA AGTCGCCCTCG; see also Materials and methods |
| Sequence-based reagent | mDia1 miRNA #1 | This paper | miRNA | CAGCATGGCT AAATGGTCA; see also Materials and methods |
| Sequence-based reagent | mDia1 miRNA #2 | This paper | miRNA | GGGTCCGTTT GCTGCCTTA; see also Materials and methods |
| Sequence-based reagent | mDia1 miRNA #3 | This paper | miRNA | GGGTAGCAAT GCTGTGTTT; see also Materials and methods |
| Sequence-based reagent | mirVana miRNA Mimic, Negative Control #1 | Thermo Fisher Scientific | Cat# 4464058 | |
| Sequence-based reagent | MYH9 sgRNA | Synthego | sgRNA | AAACUUCAUCA AUAACCCGC |
| Recombinant DNA reagent | Mouse GFP-M2A | Jordan Beach (Loyola University, Chicago) | CRISPR GFP- M2A template |
|
| Recombinant DNA reagent | Mouse Scarleti-M2A | Jordan Beach (Loyola University, Chicago) | mScarleti-CRISPR M2A template |
|
| Peptide, recombinant protein | Alt-R S.p. HiFi Cas9 Nuclease V3 |
IDT | Cat# 1081060 | |
| Peptide, recombinant protein | BAFF | R&D Systems | Cat# 8876-BF-010 | Cell culture 5 ng/ml |
| peptide, recombinant protein | Streptavidin | MilliporeSigma | Cat# 189730 | |
| peptide, recombinant protein | Mouse histidine-tagged ICAM-1 | Sino Biological | Cat# 50440-M08H | |
| Commercial assay or kit | Nucleofector Kit V | Lonza | Cat# VCA-1003 | |
| Commercial assay or kit | In-Fusion HD Cloning | Takara | Cat# 638911 | |
| Commercial assay or kit | Mini-extruder kit | Avanti Polar Lipids | Cat# 610000 | |
| Commercial assay or kit | Sticky-Slide VI0.4
Luer closed chambers |
Ibidi | Cat# 80608 | |
| Commercial assay or kit | Alexa Fluor 647 Antibody Labeling Kit |
Thermo Fisher Scientific | Cat# A20186 | |
| Commercial assay or kit | EZ-Link Micro Sulfo-NHS-Biotinylation Kit |
Thermo Fisher Scientific | Cat# 21925 | |
| Software, algorithm | ImageJ | NIH | ||
| Software, algorithm | Fiji | Schindelin et al., 2012 | RRID:SCR_002285 | https://imagej.net/Fiji |
| Software, algorithm | Softworx | Applied Precision Ltd.; GE Healthcare Life Sciences | RRID:SCR_019157 | |
| Software, algorithm | ZEN | Zeiss | RRID:SCR_018163 | |
| Software, algorithm | FibrilTool | Boudaoud et al., 2014 | RRID:SCR_016773 | |
| Software, algorithm | BLOCK-iT RNAi Designer | Thermo Fisher Scientific | RRID:SCR_002794 | https://rnaidesigner.thermofisher.com/rnaiexpress/ |
| Software, algorithm | Prism | GraphPad | RRID:SCR_002798 | |
| Software, algorithm | Traction Force plugin |
Martiel et al., 2015 |