C fos sc 52

Whole cell lysates were prepared as described previously (Xu et al., 2012 (link)). For immunoblotting analysis, lysates were separated by 10% SDS-PAGE and transferred to a PVDF membrane (Millipore) for probing with antibodies. The antibodies against p38 (sc-535) and c-Fos (sc-52) were purchased from Santa Cruz Biotechnology, YY1 antibody (ab109237) was purchased from Abcam, and c-Jun antibody (#9165) was obtained from Cell Signaling Technology.

Total cellular extracts were obtained using lysis buffer containing 150 mM Tris-HCl (pH 6.8), 6% SDS, 30% glycerol, and 0.03% Bromophenol Blue; 10% 2-ME was added immediately before harvesting cells. Cell lysates were fractionated on 7.5% SDS-PAGE, transferred to Immobilon-P membranes (Millipore), and incubated with specific antibodies. Western Lightning plus-ECL (PerkinElmer) was used for detection. NFATc1 antibody (556602, 1:1000) was from BD Biosciences; Blimp1 (sc-47732, 1:1000), c-Fos (sc-52, 1:1000), IRF8 (sc-6058, 1:1000), and p38α (sc-535, 1:3000), B-Myb (sc-390198, 1:1000) antibodies were from Santa Cruz Biotechnology; IRF1 (8478, 1:1000) antibody was obtained from Cell Signaling Technology.

Whole cell extracts and nuclear cell extracts were prepared as described [25 (link)]. Proteins were separated by SDS-PAGE and transferred onto a nitrocellulose membrane (Amersham) by semi-dry blotting. For immuno-detection, the antibodies against Δ-Actin (C4, Santa Cruz), BIM (B7929, Sigma), FasL (G247-4, BD Pharmingen), ERK2 (sc-154, Santa Cruz), γH2AX (#05-164, Upstate), XPF (Ab-5, NeoMarkers), c-Fos (sc-52, Santa Cruz) were diluted 1:1000 in 5% non-fat dry milk/Tween-TBS. For western blot analysis with phospho-specific antibodies, cells were directly lyzed in 1 x SDS-PAGE sample buffer (Roti^®-Load 1, Carl Roth GmbH) and subsequently sonified. Rabbit phospho-specific antibodies (anti-p-cJun: #3270, anti-p-JNK: #4668P; anti-p-p38K: #4511P; anti-p-ERK1/2: #4370P) as well as non- phospho-specific antibodies (anti-cJun: #9165; anti-JNK: #9258P; anti-p38K: #9212P and anti-ERK1/2: #4695P) were purchased from Cell Signaling Technology (Boston, MA, USA) and diluted 1:1000 in 5% BSA/Tween-TBS. The secondary anti-rabbit or anti-mouse antibodies (Rockland) were diluted 1:2000 in 5% non-fat dry milk/Tween-TBS.