Table 1 Antibodies used in the study
From: Cancer-associated fibroblasts actively compress cancer cells and modulate mechanotransduction
Antigen | Antibody/Chemical | Dilution | Reference | Vendor | ||||
|---|---|---|---|---|---|---|---|---|
FACS | Cryosections | Whole-mount | Cultured cells | Western blot | ||||
Fibronectin | Anti-Fibronectin rabbit polyclonal antibody | – | 1–100 | 1–100 | 1–200 | 1–10,000 | F3648 | Sigma Aldrich |
N-Cadherin | Anti-N-Cadherin mouse monoclonal antibody | – | – | – | – | 1–500 | 33-3900 | Thermo Fisher Scientific |
MyosinIIA | Anti-MyosinIIA rabbit polyclonal antibody | – | – | 1–100 | 1–100 | 1–100 | 909801 | Biolegend |
GAPDH | Anti-GAPDH rabbit polyclonal antibody | – | – | – | – | 1–5000 | G9545 | Sigma Aldrich |
αSMA | Anti-αSMA mouse monoclonal antibody | 1–200 | – | – | 1–100 | – | A2547 | Sigma Aldrich |
pMLC | Anti-pMLC rabbit polyclonal antibody | – | – | 1–100 | 1–200 | – | 3674 | Cell Signaling |
YAP | Anti-YAP rabbit monoclonal antibody | – | 1–200 | - | 1–200 | – | 14074 | Cell Signaling |
CC3 | Anti- Cleaved Caspase 3 rabbit monoclonal antibody | – | - | 1–100 | 1–200 | – | 9661 | Cell Signaling |
PHH3 | Anti-Phospho Histone H3 rabbit monoclonal antibody | – | 1– 200 | – | 1–200 | – | H0412 | Sigma Aldrich |
Ki67 | Anti-Ki67 rabbit monoclonal antibody | – | 1–200 | – | 1–400 | – | 9129S | Cell Signaling |
EpCAM | PE/Cy7 anti-human CD326, mouse IgG2b, κ | 1–10 | – | – | – | – | 324221 | Biolegend |
EpCAM | APC anti-mouse CD326, mouse, rat IgG1 | 1–10 | – | – | – | – | 130-102-969 | Miltenyi Biotec |
CD45 | Brilliant-Violet 421 anti-human CD45, mouse IgG1, κ | 1–20 | – | – | – | – | 304032 | Biolegend |
CD45 | Brilliant-Violet 421 anti-mouse CD45, rat IgG2b, κ | 1–20 | – | – | – | – | 103133 | Biolegend |
CD31 | FITC anti-human CD31, mouse IgG1, κ | 1–20 | – | – | – | – | 303104 | Biolegend |
CD31 | FITC anti-human CD31, rat IgG2a, κ | 1–20 | – | – | – | – | 102405 | Biolegend |
Vimentin | APC anti-human Vimentin, recombinant IgG1 | 1–250 | – | – | – | – | 130-106-370 | Miltenyi Biotec |
Isotype controls | FITC mouse IgG1 | 1–10 | – | – | – | – | 130-092-213 | Miltenyi Biotec |
Brilliant-Violet 421 rat IgG2b, κ | 1–20 | – | – | – | – | 400639 | Biolegend | |
FITC rat IgG2a, κ | 1–20 | – | – | – | – | 400505 | Biolegend | |
Anti-mouse Alexa Fluor 488 | Goat anti-mouse IgG Alexa Fluor 488 polyclonal antibody | – | 1–200 | 1–200 | 1–400 | – | A11029 | Thermo Fisher Scientific |
Anti-mouse Alexa Fluor 546 | Goat anti-mouse IgG Alexa Fluor 546 polyclonal antibody | – | 1–200 | 1–200 | 1–400 | – | A11030 | Thermo Fisher Scientific |
Anti-Rabbit Alexa Fluor 488 | Goat anti-Rabbit IgG Alexa Fluor 488 polyclonal antibody | – | 1–200 | 1–200 | 1–400 | – | A32731 | Thermo Fisher Scientific |
Anti-Rabbit Alexa Fluor 568 | Goat anti-Rabbit IgG Alexa Fluor 568 polyclonal antibody | – | 1–200 | 1–200 | 1–400 | – | A11011 | Thermo Fisher Scientific |
Anti-rabbit HRP | Goat anti-Rabbit polyclonal antibody | 32260 | Thermo Fisher Scientific | |||||
Against Fibronectin | – | – | – | – | 1–10,000 | |||
Against MyosinIIA | – | – | – | – | 1–5000 | |||
Against GAPDH | – | – | – | – | 1– 2500 | |||
Anti-mouse HRP | Goat anti-Mouse polyclonal antibody | 32230 | Thermo Fisher Scientific | |||||
Against N-Cadherin | – | – | – | – | 1–500 | |||
F-actin | Phalloidin-Rhodamine | – | 1–200 | 1–100 | 1–200 | – | R415 | Thermo Fisher Scientific |
Phalloidin-Alexa Fluor 488 | – | 1–200 | 1–100 | 1–200 | – | A12379 | Thermo Fisher Scientific | |
Phalloidin-Alexa Fluor 633 | – | – | – | 1–50 | A22284 | Thermo Fisher Scientific | ||
DNA | DAPI (4′,6-Diamidino-2-Phenylindole, Dihydrochloride) | – | 1–400 | 1–400 | 1–400 | – | D1306 | Thermo Fisher Scientific |
