Jnk c 17

The following antibodies were used at the indicated dilutions for immunoblotting: pJNK (#9251, 1:1000), p-p38 (#9211, 1:1000), pERK1/2 (#9101, 1:1000) and pSTAT3 antibodies (#9131, 1:1000), obtained from Cell Signaling; JNK (C-17, 1:1000), p38 (H-147, 1:1000), ERK1/2 (K-23, 1:1000), TRAF6 (H-274, 1:1000), anti-ubiquitin (P4D1, 1:1000), RAN (C-20, 1:1000), STAT3 (C-20, 1:1000), c-Myc (9E10, 1:5,000) and tubulin antibodies (10D8, 1:2000), obtained from Santa Cruz Biotech; and anti-K63 ubiquitin (HWA4C4, 1:750) and anti-Ubc13 antibodies (#371100, 1:1000), obtained from Enzo and Invitrogen, respectively. TRAF6 antibody (H-274) was also used for immunoprecipitation (1 g/sample). For ChIPs, the following antibodies were used at 2 g/sample: STAT3 (C-20) and c-Ets-1 antibodies (C-20), obtained from Santa Cruz Biotech; H3K4me3 (CS200580) and total H3 antibodies (04-928), obtained from Millipore; and Set1 antibody (A300-289A) from Bethyl Laboratories. For flow cytometry, fluorescently labeled antibodies to CD11b (M1/70, FITC-labeled), F4/80 (BM8, PerCP Cy5.5-labeled), M-CSFR (AFS98, APC-labeled) and RANK (R12-31, PE-labeled) from eBioscience were used at a 1:100 dilution. Recombinant murine RANKL, IL-6, IL-10 and IL-6 blocking antibody were purchased from R&D Systems.