Table 2.
Cytokines, inhibitors, and antibodies used in this study.
| Concentration | Source | ||
|---|---|---|---|
| Antibodies | |||
| mouse anti-ACTB | WB | 1:10,000 | Sigma-Aldrich, Schnelldorf, Germany, CAT. A5441 |
| rabbit anti-CLDN1 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA Cat. 519000 |
| rabbit anti-CLDN2 (used for mouse tissue analysis) | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA Cat. 516100 |
| mouse anti-CLDN2 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 325600 |
| rabbit anti-CLDN3 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 341700 |
| mouse anti-CLDN4 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 329400 |
| mouse anti-CLDN7 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 349100 |
| rabbit anti-CLDN8 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA Cat. 400700Z |
| rabbit anti-CLDN15 | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 364200 |
| rabbit anti-ILDR1 | WB | 1:1000 | Bioss, ThermoFisher, Waltham, MA, USA, Cat: bs-11013R |
| rabbit anti-LSR | WB | 1:1000 | Atlas antibodies, Cat. HPA007270 |
| rabbit anti-MD3 | IF WB |
1:200 1:1000 |
Proteintech, Rosemont, IL, USA, Cat. 25567-1-AP |
| rabbit anti-MD3 (used for patient tissue and some mouse tissue analysis) | WB | 1:1000 | Kind gift of Jerrold Turner, Laboratory of Mucosal Barrier Pathobiology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA, [13,14] |
| rabbit anti-OCCL | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 711500 |
| rabbit anti-TRIC | WB | 1:1000 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. 700191 |
| mouse anti-TUBA | WB | 1:4000 | Sigma-Aldrich, Schnellendorf, Germany, CAT. T9026 |
| mouse anti-ZO-1 647 | IF | 1:500 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. MA3-39100-A647 |
| DAPI | IF | 1:1000 | Roche Diagnositics, Mannheim, Germany, Cat. 10 236 276 001 |
| goat anti-rabbit IgG (H+L) secondary antibody Alexa Flour 488 | IF | 1:500 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. A11034 |
| goat anti-mouse IgG (H+L) antibody Alexa Flour 594 | IF | 1:500 | Invitrogen, ThermoFisher, Waltham, MA, USA, Cat. A11032 |
| secondary peroxidase-conjugated antibodies anti-rabbit | WB | 1:10000 | Jackson ImmunoResearch, Cambridge House, UK, Cat. 111-036-003 |
| secondary peroxidase-conjugated antibodies anti-mouse | WB | 1:10000 | Jackson ImmunoResearch, Cambridge House, UK, Cat. 115-036-003 |
| Cytokines | |||
| TNFα | 1000 | U/mL | PeproTech, Hamburg, Germany |
| IFNγ | 50 | U/mL | PeproTech, Hamburg, Germany |
| IL-1α | 10 | ng/mL | PeproTech, Hamburg, Germany |
| TGFβ 1 | 10 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| TGFβ 2 | 10 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| IL-4 | 100 | ng/mL | PeproTech, Hamburg, Germany |
| IL-5 | 50 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| IL-6 | 50 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| IL-9 | 100 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| IL-13 | 100 | ng/mL | PeproTech, Hamburg, Germany |
| IL-22 | 100 | ng/mL | Miltenyi Biotec, Bergisch Gladbach, Germany |
| Inhibitors | |||
| U0126 | 10 | µM | Cell signaling Technology, Frankfurt am Main, Germany |
| SAHA | 5 | µM | Sigma-Aldrich, Schnelldorf, Germany |
| AG490 | 50 | µM | Sigma-Aldrich, Schnelldorf, Germany |
| Tanshinone IIa | 10 | µM | Sigma-Aldrich, Schnelldorf, Germany |
| STAT3-Inhibitor VI | 10 | µM | Calbiochem, Darmstadt, Germany |
| JAK3-Inhibitor II | 50 | µM | Calbiochem, Darmstadt, Germany |
| JNK V | 10 | µM | Calbiochem, Darmstadt, Germany |
| SP600125 | 10 | µM | Cell Signaling Technology, Frankfurt am Main, Germany |