Nintedanib bibf 1120

Nintedanib (BIBF 1120) and docetaxel were obtained from Selleck Chemicals LLC (Houston, TX, USA). Silibinin was purchased from Sigma-Aldrich (St. Louis, MO, USA). All reagents were dissolved in sterile dimethylsulfoxide (DMSO) to prepare 10 mmol/L stock solutions, which were stored in aliquots at −20 °C until use. Working concentrations were diluted in culture medium prior each experiment. Antibodies against total STAT3 (124H6, Cat. No 9139), phospho-STAT3 Tyr705 (D3A7, Cat. No 9145S), and phospho-STAT3 Ser727 (Cat. No 9134) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against β-actin (Clone #2D4H5, Cat. No 66009-1-Ig,) and human recombinant human IL-6 (Cat. HZ-1019-10) were purchased from Proteintech Group, Inc., Rosemont, IL, USA). Lysotracker^® Red DND-99 (Cat. No L7528) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Ammonium chloride was purchased from Sigma-Aldrich (Cat. No A9434). Bafilomycin A1 was obtained from Calbiochem (Cat. No 196000).

Isogenic pairs of vemurafenib‐sensitive and vemurafenib‐resistant melanoma cells (M229, M238, M249) were provided by R. Lo. UACC62 vemurafenib‐sensitive (UACC62P) and vemurafenib‐resistant melanoma cells (UACC62R) were provided by Neubig's laboratory. 1205Lu cells were obtained from Rockland. YUMM1.7 mouse melanoma cells were a kind gift from M. Bosenberg (Meeth et al, 2016 (link)). A375DR melanoma cells were provided by S. Shen. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 7% FBS (Hyclone) and 1% penicillin/streptomycin. Resistant cells were continuously exposed to 1 μM of vemurafenib. Cell lines were routinely tested for the absence of Mycoplasma by PCR.
Short‐term cultures of patient melanoma cells MM034 and MM099 were generated in the laboratory of Pr G. Ghanem. Culture reagents were purchased from Thermo Fisher Scientific. Vemurafenib (PLX4032), trametinib (GSK1120212), cobimetinib (SB431542, GSK690693), nintedanib (BIBF1120), and staurosporine were from Selleckchem. CP673451 was purchased from Tocris Bioscience. Recombinant human TGF‐β1 was obtained from ImmunoTools. Recombinant human PDGF‐BB was purchased from Peprotech.
Information on all reagents used is provided in Appendix Tables S4–S6.