C ebpβ c terminal

Chromatin from BMM was analyzed using a modification of the ChIP
Millipore/Upstate protocol (MCPROTO407) as described^{(29 (link))} using Magna ChIP Protein A+G Beads
(16-663, Millipore). The following antibodies were used for ChIP: EZH2 (Active
Motif, 39902), H3K27me3 (Active Motif, 39155), C/EBPβ C-terminal (Santa
Cruz, sc-150) and C/EBPβ-LAP (CST, 3087). Aliquots for input and
non-specific IgG control samples were included with each experiment.
ChIP-qPCR primers are listed in Table 2. Fold enrichment was calculated based on Ct
as 2^(ΔCt), where ΔCt = (Ct_Input –
Ct_Ip). The IgG ΔCt was subtracted from the specific Ab
ΔCt to generate ΔΔCt = (ΔCt_{specific Ab}– ΔCt_IgG).

Immunoblotting was performed according to standard protocols using
following antibodies directed towards: EZH2 (CST, 3147), H3K27me3 (Active Motif,
39155), H3 (CST, 9715), H3K9ac (Active Motif, 61251), β-actin (Sigma,
85316), MafB (Santa Cruz, sc-376387), C/EBPβ C-terminal (Santa Cruz,
sc-150), C/EBPβ-LAP (CST, 3087), pPI3K-p85(Tyr458)/p55(Tyr199) (CST,
4228), pPDK1-Ser241 (CST, 3061), AKT (CST, 9272S), pAKT-Ser473 (CST, 4060), mTOR
(CST, 2983), pmTOR Ser2448 (CST, 2971), and pS6RP-Ser235/236 (CST, 2211)..
Nuclear and cytoplasmic protein separations were performed using the Nuclear
Extract Kit (Active Motif, 40010). Band signals were detected using Amersham ECL
Wstern Blotting Detection Reagent (GE Life Sciences, RPN2106) and analyzed and
quantitated using ProteinSimple Imager and AlphaView software (ProteinSimple,
CA).