Anti cebpa 14aa

CoIP was performed as described previously (M€ uller et al., 2010) . Anti-C/EBPa (14AA; Santa Cruz Biotechnology), anti-FLAG (M2, F3165; Sigma-Aldrich), anti-HA (MMS-101R; Convace), and anti-Tip60 (NBP2-20647; Novus Biologicals) were used for precipitation as indicated. To detect the acetylation of C/EBPa in Fao cells, endogenous level, or in transiently transfected HEK293T cells, the cells were treated with the deacetylase inhibitors 1 mM TSA (T8552; Sigma-Aldrich) and 5 mM NAM (47865U; Sigma-Aldrich) 8 hr before harvesting. The IP lysis buffer and IP wash buffer were supplemented with these inhibitors as well.

The pcDNA3-based full-length (p42) rat C/EBPa and rat C/EBPa-FLAG have been described earlier (M€ uller et al., 2010) ; cloning details are available upon request. See Supplemental Experimental Procedures for other plasmids used.
Cell Culture, Transfection, and Immunofluorescence All cells were cultured in DMEM plus 10% fetal calf serum (FCS) (Invitrogen) and penicillin/streptomycin at 5% CO 2 and 37 C. HEK293T cells were seeded at 2.5 3 10 6 cell in 10 cm dishes and transfected the next day with 5 mg expression vectors using calcium phosphate. The immunofluorescence staining protocol was described previously (M€ uller et al., 2010) . The primary antibodies used were anti-C/EBPa (14AA; Santa Cruz Biotechnology), anti-FLAG (M2, F3165; Sigma-Aldrich), and anti-HA (MMS-101R; Convace). Secondary antibodies used were Alexa Fluor 488 or 568 conjugated (Invitrogen). p300 inhibitor C646 (CAS 328968-36-1; Sigma-Aldrich) was used at final concentration of 10 mM.
(D and F) Basal and maximal ECAR in cumate-induced Hepa1-6 cells expressing WT, K159/298Q-, or K159/298R-C/EBPa-FLAG proteins and cultured in medium with 25 mM (D) or 2.5 mM (F) glucose. For all experiments (n = 5), statistical differences were analyzed using Student's t tests. Error bars represent ± SD. *p < 0.05, **p < 0.01, ***p < 0.001; NS, not significant.