Dynamic Acetylation of All Lysine 4–Methylated Histone H3 in the Mouse Nucleus: Analysis at c-fos and c-jun
Figure 7
Effect of TSA Pre-Treatment on TPA-Stimulated MAP Kinase Activation and Transcription Factor Phosphorylation
(A) Quiescent C3H 10T½ cells were treated with TSA (10 or 500 ng/ml; 5 min to 4 h). Positive controls for MAP kinase activation included ERK1/2 (TPA [T]; 10 min), JNK/SAPKs, and p38 (sAn; 30 to 60 min). “C” indicates control (unstimulated). Cell extracts were analysed by Western blotting with anti-ERK1/2, anti-phospho-p38, and anti–ACTIVE JNK antibodies. The mobility of ERKs is retarded on activation. Activation of p38 and JNK/SAPK results in phosphorylation. Note that anti–ACTIVE JNK also recognises activated ERK1/2 (lane 14).
(B) Quiescent C3H 10T½ cells were untreated (−) or pre-treated with TSA (10 or 500 ng/ml; 15 min). Cells were then left unstimulated (C) or stimulated with TPA for 5 to 30 min. Cell extracts were analysed by Western blotting with anti-ERK1/2 antibody.
(C) Quiescent C3H 10T½ cells were untreated (−) or pre-treated with TSA (500 ng/ml; 15 min). Cells were stimulated with TPA for 15 to 30 min. Cell extracts were analysed by Western blotting with anti-ATF-2 and anti-phospho-CREB antibodies. Phosphorylation of ATF-2 results in retarded mobility.