Tpca 1

Recombinant mouse EGF was purchased from PeproTech, Inc. Recombinant human IL-1β and TNF-α were obtained from Miltenyi Biotec, GmbH (Bergisch Gladbach). The MEK inhibitors U0126 and PD98059, and the EGFR inhibitor PD153035 were purchased from Calbiochem (Merck KGaA). The NF-κB inhibitor BAY 11-7085 was obtained from Cayman Chemical. Recombinant human FGF-1 and the NF-κB kinase-2 (IKK-2) inhibitor TPCA-1 were purchased from R&D Systems, Inc. The FGFR1 inhibitor SU-5402 was obtained from Wako Pure Chemical Industries, Ltd. We confirmed that dimethyl sulfoxide (DMSO), the vehicle used for the U0126, PD98059, PD153035, BAY 11-7085, TPCA-1, and SU-5402 treatments, did not affect the expression of the MF markers α-SMA and type I collagen (data not shown). Heparin sodium salt was obtained from Merck KGaA. Heparin was included to achieve the optimal FGF-1 activity (28 (link)).

Cells were plated at 3 × 10⁶ cells/ml in a 24‐well plate (500 μl) for western blotting and at 1.33 × 10⁶ cells/ml in a 96‐well plate (150 μl) for ELISA and flow cytometry. Stimuli were used at the following concentrations, if not indicated otherwise: 20 μg/ml 2′‐3′‐cGAMP (synthesized in‐house, detailed synthesis instructions available upon request) for ELISA and WB, and 40 μg/ml 2′‐3′‐cGAMP for flow cytometry; 5:1 cells:beads αCD3/CD28 (Gibco, #11132D), 1 μM ABT737 (Selleck Chemicals, #S1002), 1 μM S63845 (Selleck Chemicals, #S8383), 6,000 IU/ml IFNα2a (Miltenyi Biotec, #130‐093‐874), 20 μM zVAD (R&D Systems, #FMK001), 2 μg/ml PHA (Sigma‐Aldrich, #61764), 50 ng/ml PMA (Enzo Life Sciences, #BML‐PE160‐0005), 1 IU/ml ionomycin (Alomone Labs, #I‐700), 10 ng/ml IFNγ (PeproTech, #300‐02), 10 ng/ml TNFα (PeproTech, #300‐01A), 50 IU/ml IL2 (R&D Systems, #202‐IL‐010), TPCA‐1 (R&D Systems, #2559/10), Dasatinib (Selleck Chemicals, #S1021), Tacrolimus (Astellas PHArma, #06896463), Torin1 (InvivoGen, #inh‐tor1), and 2 μΜ Thapsigargin (Sigma‐Aldrich, #T9033‐.5MG). For HT‐DNA delivery, 200 ng of HT‐DNA (Sigma‐Aldrich, #D6898‐1G) per condition was complexed with 0.5 μl of Lipofectamine 2000 (Thermo Fisher Scientific, #11668019) per condition in Opti‐MeM (Gibco, #10149832) for 5 min at RT.