Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | RTN1A | NM_0211369, Eurofins (this paper) |
||
| Cell line (Homo sapiens) | THP-1 | ATCC | TIB-202 | |
| Transfected construct (Homo sapiens) | pHR‐SIN‐BX‐IRES‐Emerald | Paster et al., 2013 | Lentiviral construct to transfect the THP-1 cell line and express RTN1A |
|
| Antibody | α-RTN1A (clone: mon162), unconjugated (Mouse monoclonal) |
abcam | ab9274 | IF staining – 0.2 µg/ml; cultivation of epidermal sheets – 5 µg/ml/ |
| Antibody | α-RTN1A (clone: mon162), unconjugated (Mouse monoclonal) |
Novus Biologicals, Biotechne | NBP1-97677 | IF staining – 0.2 µg/ml; cultivation of epidermal sheets – 5 µg/ml/ |
| Antibody | α-RTN1A-APC (clone: mon161, Mouse monoclonal) |
Novus Biologicals, Biotechne | NBP1-97678AF647 | FACS analysis – 1:500 |
| Antibody | IgG1 [15-6E10A7] - Isotype control, unconjugated (Mouse monoclonal) |
abcam | ab170190 | IF staining – 0.2 µg/ml; cultivation of epidermal sheets – 5 µg/ml/ |
| Antibody | α-CD207-FITC (clone 929F3.01, Rat monoclonal) |
Dendritics | Cat:DDX0362 | IF staining – 0.25 µg/ml. 1:250 |
| Antibody | α-CD207, unconjugated (Rabbit monoclonal) |
Sigma-Aldrich | HPA011216 | IF staining – 1:200 |
| Antibody | α-Human CD207- PE (clone DCGM4, Mouse) |
Beckman Coulter | PN IM3577 | FACS analysis – 1:200 |
| Antibody | α-Human HLA- DR-PE (Mouse) |
BD Pharmingen | Cat. 347401 | FACS analysis – 1:100 |
| Antibody | α-Human CD1a- PE (Mouse) |
BD Pharmingen | Cat. 555807 | FACS analysis – 1:100 |
| Antibody | α-Human CD83- FITC (Mouse) |
BD Pharmingen | Cat. 556910 | FACS analysis, IF staining – 1:100 |
| Antibody | α-Human CD86- FITC (Mouse) |
BD Pharmingen | Cat. 555657 | FACS analysis, IF staining – 1:100 |
| Antibody | α-CCR7-APC | Miltenyi Biotech | 5171113456 | FACS analysis, IF staining – 1:100 |
| Antibody | Recombinant α- Vimentin-FITC (clone REA409) |
Miltenyi Biotec | 130-116-508 | IF staining – 1:50 |
| Antibody | Recombinant FITC isotype control |
Miltenyi Biotech | 130-113-449 | IF staining – 1:50 |
| Antibody | F(ab')2-Goat anti- Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 |
Life Technologies | REF A-11070 | IF staining – 1:500 |
| Antibody | F(ab')2-Goat anti- Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 |
Invitrogen, Thermo Fisher Scientific | REF A-11018 | IF staining – 1:500 |
| Antibody | Alexa Flour 647F(ab´) 2 fragment of goat α-rabbit IgG (H+L) |
Invitrogen, Thermo Fisher Scientific | REF A-21246 | IF staining – 1:500 |
| Commercial assay or kit | Human TLR kit | InvitroGen | Cat. tlrl-kit1hw | |
| Commercial assay or kit | LEGENDplex Human inflammatory Panel 1 (13-plex) w/VbP |
Biolegend | Cat. 740809 | |
| Commercial assay or kit | LEGENDplex Human Adhesion Molecule Panel (13-plex) w/VbP |
Biolegend | Cat. 740946 | |
| Chemical compound, drug | Thapsigargin | Invitrogen, Thermo Fisher Scientific | T7458 | |
| Software, algorithm | GraphPad Prism v8.0.1 | GraphPad Software | ||
| Software, algorithm | FlowJo v10.6.1 | BD (Becton, Dickinson & Company) | ||
| Software, algorithm | Fiji: ImageJ | Schindelin et al., 2012 | ||
| Software, algorithm | ImarisViewer 9.8 | Oxford Instruments | ||
| Software, algorithm | Adobe Illustrator CS7 | Adobe Inc | ||
| Other | Alexa Fluor 647 Phalloidin |
Invitrogen | REF A-22287 | IF staining – 1:500 |
| Other | Fixable Viability Dye eFluor 450 |
eBioscience | Cat. 65-0863-18 | FACS analysis – 1:1000 |
| Other | DAPI (4′,6-diamidino- 2-phenylindole dihydrochloride) |
Sigma-Aldrich | Cat. D9542 | IF staining – 1:5000 |
| Other | Dispase II (neutral protease, grade II) |
Roche Diagnostics | 04942078001 | 1.2 U/ml |
| Other | ProLong Gold antifade reagent |
InvitroGen | Cat. P36934 | |
| Other | IC Fixation Buffer | eBioscience | Cat. 00-8222-49 | |
| Other | Permeabilization Buffer (×10) | eBioscience | Cat. 00-8333-56 | |
| Other | Phorbol 12-myristate 13-acetate (PMA), PKC activator |
Abcam | Ab120297 | |
| Other | Triton X-100 | Sigma-Aldrich | T9284 | |
| Other | Fura Red, AM, cell permeant |
Invitrogen, Thermo Fisher Scientific | F3020 | |
| Other | CellTrace CFSE Cell Proliferation Kit, for flow cytometry |
Invitrogen, Thermo Fisher Scientific | C34554 | |
| Other | Ionomycin- Calcium ionophore – NFAT Activator |
InvivoGen | inh-ion | |
| Other | Fibronectin Solution Human | PromoCell | Cat. C-43060 |