Anti fli1

Antibodies used for Western blotting included anti-Myc, anti-Flag, anti-HA, anti-pJNK, anti-JNK, anti-pIKK, anti-TAK1, anti-pTAK1, cleaved caspase 3, and anti-β-tubulin from Cell Signaling. Anti-Fli1 and anti-iNOS were purchased from Santacruz Biotechnology, and anti-TRB3 antibodies were a gift from Dr Marc Montminy, Salk Institute. For immunofluorescence, mouse anti-BAX clone 6A7 (BD Bioscience), sheep anti-insulin (Binding Site), and In Situ Cell Death Detection Kit (Fluorescein) from Roche were purchased from commercial sources. Agarose-conjugated antibodies used for immunoprecipitations included anti-HA agarose (Thermo Fisher Scientific) and anti-FlagM2 agarose (Sigma-Aldrich). Streptavidin-Agarose was purchased from Pierce. Immunofluorescence and Western blotting employed fluorescent or HRP-conjugated secondary antibodies (Jackson ImmunoResearch), the latter detected using Supersignal West-Pico Plus chemiluminescence reagent (Thermo Fisher). Other reagents include IL-1β (Peprotech) and ultrapure TLR4-specific LPS (InvivoGen). Flag-TRB3 adenovirus was generated as described 3). Smartpool TARGETplus siRNA was used to knockdown Fli1, and nontargeting siRNA was used as control (Dharmacon-Horizon Discovery Sciences). TPCA-1 (Bio-Techne-Tocris) was used to inhibit the NFκB pathway, and MLK3 was inhibited using CEP11004 (gift from Cephalon Inc).