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. Author manuscript; available in PMC: 2013 Aug 17.
Published in final edited form as: Cell. 2012 Aug 17;150(4):685–696. doi: 10.1016/j.cell.2012.07.018

Figure 1. EGF-Induced and PKM2-Dependent Phosphorylation of Histone H3 at T11 Is Required for Acetylation of Histone H3 at K9.

Figure 1

Please also see supplemental Figures S1-S4. Immunoprecipitation and immunoblotting analyses were performed with the indicated antibodies.

(A) U87/EGFR cells expressing Flag-tagged WT H3, H3-K4R, and K9R were treated with or without EGF (100 ng/ml) for 6 h.

(B, F) U87/EGFR and U251 cells expressing a control or PKM2 shRNA were treated with or without EGF (100 ng/ml) for 6 h. Endogenously expressed histone H3 was examined. Data represent the mean ± SD of three independent experiments (F).

(C) U87/EGFR were treated with or without EGF (100 ng/ml) or 20% serum with calyculin A (25 nM) for 6 h.

(D) U87/EGFR cells with or without expressing PKM2 shRNA were treated with or without EGF (100 ng/ml) for 6 h. Endogenously expressed histone H3 was immunoprecipitated.

(E) U87/EGFR cells expressing Flag-tagged WT H3, H3-T3A, H3-T6A, and H3-T11A were treated with or without EGF (100 ng/ml) for 6 h.

(G) U87/EGFR cells expressing Flag-tagged WT H3 or H3-T11A were treated with or without EGF (100 ng/ml) for 6 h.

(H) U87/EGFR cells expressing a control shRNA or shRNA against Chk1, DAPK3, or PKN1 mRNA were analyzed by immunoblotting analysis with the indicated antibodies.

(I) U87/EGFR cells expressing a control shRNA or shRNA against Chk1, DAPK3, or PKN1 mRNA were treated with or without EGF (100 ng/ml) for 6 h and analyzed by immunoblotting analysis with the indicated antibodies.